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WO2019129144A1 - 一种用于检测食道肿瘤和/或胃肿瘤良恶性程度的分级模型及其应用 - Google Patents

一种用于检测食道肿瘤和/或胃肿瘤良恶性程度的分级模型及其应用 Download PDF

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WO2019129144A1
WO2019129144A1 PCT/CN2018/124373 CN2018124373W WO2019129144A1 WO 2019129144 A1 WO2019129144 A1 WO 2019129144A1 CN 2018124373 W CN2018124373 W CN 2018124373W WO 2019129144 A1 WO2019129144 A1 WO 2019129144A1
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imprinted
imprinted gene
gene
genes
expression amount
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成彤
周宁
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Lisen Imprinting Diagnostics Inc
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Lisen Imprinting Diagnostics Inc
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer

Definitions

  • the present disclosure relates to the field of biotechnology, and relates to the field of genetic diagnosis, such as the field of genetic diagnosis, such as a hierarchical model and its application, such as a hierarchical model for detecting the degree of benign and malignant esophageal tumors and/or gastric tumors, and applications thereof, for example, A device and application of a hierarchical model of imprinted genes in detecting the degree of benign and malignant esophageal tumors and/or gastric tumors and their composition.
  • Gastric cancer and esophageal cancer are two common types of digestive tract cancers. In 2012, there were 952,000 new cases of gastric cancer, 723,000 deaths, 456,000 new cases of esophageal cancer, and 400,000 deaths. China is a high-risk area for gastric cancer and esophageal cancer. The annual incidence of new cases and deaths of gastric cancer and esophageal cancer account for about 50% of the world. The 5-year survival rate of early gastric cancer and esophageal cancer is over 90%, while the 5-year survival rate of advanced gastric cancer and esophageal cancer is less than 30%. Therefore, early diagnosis of gastric cancer and esophageal cancer is very important to save patients' lives. However, gastric cancer and esophageal cancer usually have no obvious symptoms in the early stage. The detection rate in China is less than 10%. More than half of the patients are already in the advanced stage when they find gastric cancer or esophageal cancer.
  • intraepithelial neoplasia is a precancerous lesion of the stomach, which can be classified into low-grade intraepithelial neoplasia and high-grade intraepithelial neoplasia depending on the degree of hyperplasia.
  • Low-grade intraepithelial neoplasia is about 20% likely to develop gastric cancer, while more than 70% of high-grade intraepithelial neoplasia may develop into gastric cancer.
  • the pathological distinction between low-grade and high-grade intraepithelial neoplasia is mainly subjective judgment by pathologists based on morphology.
  • CN 106609037 A discloses a detection kit for diagnosing gastric cancer patients based on a plurality of genes, wherein the kit includes a primer for diagnosing gastric cancer, which can be used for diagnosing gastric cancer, but the detection thereof is not accurate enough to distinguish which specific gastric cancer is specific. stage.
  • the pathological diagnosis of benign and malignant cells by traditional pathology is based on the relationship between cell size, morphology, invasiveness and surrounding cellular tissues. It has great limitations on the discovery of early changes in cells (cancer), so the method of cancer diagnosis at the cellular level has once become a research hotspot. With the continuous research in the field of molecular biology, more and more molecular detection techniques are applied to cancer diagnosis.
  • the production of cancer is an uncontrolled cell growth/splitting caused by epigenetic changes and genetic variations that accumulate over time.
  • Traditional pathological diagnosis is based on the size, morphology and structural variation of cells and tissues, thereby making a benign and malignant judgment of esophageal and gastric tumors.
  • molecular detection techniques With the development and deepening of molecular biology, more and more molecular detection techniques have been applied to the detection of esophageal cancer and gastric cancer. From the analysis of the development of cancer, molecular changes (epigenetics and genetics) are much earlier than changes in cell morphology and tissue structure. Therefore, molecular biology testing is more sensitive to early detection of cancer.
  • the current diagnosis of esophageal cancer and gastric cancer requires a new detection system and detection model.
  • the molecular markers of esophageal cancer and gastric cancer at the cellular level are analyzed to provide more accurate pre-diagnosis and Diagnostic information.
  • the present disclosure provides an imprinted gene grading model and its application.
  • the present disclosure provides an imprinted gene grading model for esophageal tumors and/or gastric tumors, the model calculating the amount of imprinted gene deletion expression, the imprinted gene copy number abnormal expression amount, and the total expression amount of the imprinted gene Changes in the esophageal tumor and/or gastric tumor classify the expression status of the imprinted gene;
  • the imprinted gene is any one or a combination of at least two of Z1, Z11 or Z16, the imprinting gene Z1 is Gnas, the imprinting gene Z11 is Grb10, and the imprinting gene Z16 is Snrpn/Snurf.
  • the imprinted (trace) deletion refers to the activation (demethylation) of the allele in the imprinted (spot) gene that was originally in a silent state, and is the most common and early epigenetic change in cancer, and This property can be used as a pathological marker.
  • the proportion of imprinted deletions is very low, and the imprinted gene and the imprinted gene are simultaneously a concept, indicating the same meaning, and can be replaced.
  • the public has found that the sensitivity of the diagnosis of esophageal cancer can reach 90.4% by calculating the amount of imprinted gene deletion and the abnormal expression of imprinted gene copy number in any esophageal tumor and/or gastric tumor of Z1, Z11 and Z16. Above, the sensitivity of diagnosis to gastric cancer can reach 67.6% or more.
  • any one of Z1, Z11 and Z16 can be detected if a preliminary detection detects only one imprinted gene.
  • the most preferred imprinting gene for detecting esophageal cancer is Z1.
  • the most preferred imprinting gene for detecting gastric cancer is Z11.
  • the inventors found that the detection of a Z1 imprinted gene alone can be 96.3% sensitive to esophageal cancer, and a Z11 imprinted gene can be detected alone, and the sensitivity for diagnosis of esophageal cancer can reach 92.5%. Detection of a Z16 imprinted gene, the sensitivity of diagnosis of esophageal cancer can reach 90.4%.
  • the inventors found that the detection of a Z1 imprinted gene alone can achieve a sensitivity of 80.6% for gastric cancer, and a Z11 imprinted gene can be detected alone, and the sensitivity for diagnosis of gastric cancer can reach 88.9%.
  • the Z16 imprinted gene has a sensitivity of 67.6% for the diagnosis of gastric cancer.
  • the combination may be any two of Z1, Z11 and Z16, for example the combination may be a combination of Z1 and Z11, Z1 and Z16 Combination or combination of Z11 and Z16.
  • the inventors have found that by calculating the amount of imprinted gene deletion and the abnormal expression level of imprinted gene copy number of two or more imprinted genes, the sensitivity can be further improved, and the combination of two imprinted genes of the imprinted gene can be detected to diagnose esophageal cancer.
  • Sensitivity can reach more than 98.1%.
  • the sensitivity of diagnosis to esophageal cancer can reach 98.1%.
  • the sensitivity of diagnosis to esophageal cancer can reach 99.2%.
  • the sensitivity of diagnosis to esophageal cancer can reach 99.2%;
  • the inventors have found that by calculating the amount of imprinted gene deletion and the abnormal expression of imprinted gene copy number of two or more imprinted genes, the sensitivity can be further improved, and the combination of two imprinted genes of the imprinted gene can be detected, which is sensitive to the diagnosis of gastric cancer.
  • the degree can reach more than 94.1%.
  • the sensitivity of diagnosis to gastric cancer can reach 97.1%.
  • the sensitivity of diagnosis to gastric cancer can reach 97.2%, and the detection of Z11 and Z16 is detected.
  • the diagnostic sensitivity to gastric cancer can reach 94.1%.
  • the imprinting gene further comprises any one or a combination of at least two of Z3, Z4, Z5, Z6, Z8, Z10 or Z13; wherein the imprinting gene Z3 is Peg10, the imprint The gene Z4 is Igf2r, the imprinted gene Z5 is Mest, the imprinted gene Z6 is Plagl1, the imprinted gene Z8 is Dcn, the imprinted gene Z10 is Gatm, and the imprinted gene Z13 is Sgce.
  • the method for calculating the imprinted gene is: calculating a combination of imprinted genes, the imprinted genes are ten of Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 A combination of genes.
  • the imprinted gene is deleted after the cells are subjected to hematoxylin staining, and there are two red/brown marks in the nucleus, and the abnormal copy number of the imprinted gene is that after the cells are subjected to hematoxylin staining, there are two or more red/brown marks in the nucleus.
  • the copy number abnormality is caused by abnormal gene replication of cancer cells, resulting in the expression of this gene as triploid or even higher polyploid.
  • the hematoxylin-stained label is selected from, but not limited to, red or brown, and staining with other colors can also be used for calculation of the total expression level of the imprinted gene, the amount of imprinted gene deletion, and the abnormal expression level of the imprinted gene copy number.
  • the formula for calculating the total expression amount of the imprinted gene, the expression amount of the imprinted gene, and the abnormal expression amount of the imprinted gene are as follows:
  • Imprinted gene deletion gene expression level (LOI) c / (b + c + d) ⁇ 100%;
  • a is a cell nucleus in which there is no label in the nucleus and no imprinted gene is expressed after the hematoxylin staining of the cell; and b is a red/brown mark in the nucleus after the hematoxylin staining of the cell, and the imprinting gene exists.
  • the nucleus; the c is a hematoxylin staining of the cells, there are two red/brown marks in the nucleus, and the nucleus of the imprinted gene is deleted; and the d is a hematoxylin staining of the cells, and there are more than two red/brown marks in the nucleus.
  • imprinted gene copy number abnormal cell nucleus is a cell nucleus in which there is no label in the nucleus and no imprinted gene is expressed after the hematoxylin staining of the cell
  • b is a red/brown mark in the nucleus after the hematoxylin staining of the cell,
  • the hematoxylin-stained label is selected from, but not limited to, red or brown, and staining with other colors can also be used for imprinting gene deletion expression, imprinted gene copy number abnormal expression, and imprinted gene total expression. The calculation of the amount.
  • the probe is amplified by in situ hybridization, and Hemotoxy (hematoxylin) nuclei are used to amplify the signal, and under the 40 ⁇ or 60 ⁇ microscope, the presence of the imprinted gene in each nucleus, the imprinted gene is deleted, or The copy number is abnormal, and the degree of benign and malignant tumors of the sample is determined by calculating the expression level of the imprinted gene deletion gene, the abnormal expression level of the imprinted gene copy number, and the total expression level of the imprinted gene; since the slice is only 10 ⁇ m, the nuclei seen under the microscope are approximately 20% are incomplete nuclei, which means that there is a possibility of partial false negatives.
  • Hemotoxy hematoxylin
  • the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the imprinted gene total expression amount are divided into five different levels.
  • At least 1200 cells are counted by each probe in the region where the sample is most positive, for ten of Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16
  • the imprinted gene's imprinted gene deletion expression amount, imprinted gene copy number abnormal expression amount, and imprinted gene total expression amount were divided into five different levels.
  • the five different levels of the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount of Z1 are:
  • the imprinted gene deletion expression amount of the imprinted gene Z1 is less than 16%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z1 is less than 2.5%, or the total expression amount of the imprinted gene Z1 is less than 35% One or a combination of at least two;
  • the imprinted gene deletion expression amount of the imprinted gene Z1 is 16-20%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z1 is 2.5-4%, or the total expression amount of the imprinted gene Z1 is 35 Any one of -40% or a combination of at least two;
  • the imprinted gene deletion expression amount of the imprinted gene Z1 is 20-25%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z1 is 4-6.5%, or the total expression amount of the imprinted gene Z1 is 40. -50% of any one or a combination of at least two;
  • the imprinted gene deletion expression amount of the imprinted gene Z1 is 25-30%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z1 is 6.5-8%, or the total expression amount of the imprinted gene Z1 is 50 -65% of any one or a combination of at least two;
  • Grade IV the imprinted gene deletion expression amount of the imprinted gene Z1 is greater than 30%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z1 is greater than 8%, or the total expression amount of the imprinted gene Z1 is greater than 65% One or a combination of at least two.
  • the five different levels of the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount for Z3 and Z13 are:
  • the imprinted gene deletion expression amount of the imprinted genes Z3 and Z13 is less than 10%, the imprinted gene copy number abnormal expression amount of the imprinted genes Z3 and Z13 is less than 1% or the total expression amount of the imprinted genes Z3 and Z13 Any one of less than 25% or a combination of at least two;
  • the imprinted gene Z3 and Z13 have an imprinted gene deletion expression of 10-15%, the imprinted genes Z3 and Z13 have an imprinted gene copy number abnormal expression of 1-2% or the imprinted genes Z3 and Z13
  • the total expression amount is any one of 25-30% or a combination of at least two;
  • the imprinted gene Z3 and Z13 have an imprinted gene deletion expression of 15-25%, the imprinted genes Z3 and Z13 have an imprinted gene copy number abnormal expression of 2-3% or the imprinted genes Z3 and Z13
  • the total expression amount is any one of 30-45% or a combination of at least two;
  • the imprinted gene Z3 and Z13 have an imprinted gene deletion expression of 25-30%, the imprinted genes Z3 and Z13 have an imprinted gene copy number abnormal expression of 3-5% or the imprinted genes Z3 and Z13
  • the total expression amount is any one of 45-55% or a combination of at least two;
  • Grade IV the imprinted gene deletion expression amount of the imprinted genes Z3 and Z13 is greater than 30%, the imprinted gene copy number abnormal expression amount of the imprinted genes Z3 and Z13 is greater than 5% or the total expression amount of the imprinted genes Z3 and Z13 Any one of more than 55% or a combination of at least two;
  • the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount of the imprinted genes Z3 and Z13 are independent of each other.
  • the five different levels of the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount for Z4, Z5, Z6, Z10, and Z11 are:
  • the imprinted gene Z4, Z5, Z6, Z10 and Z11 have an imprinted gene deletion expression of less than 16%, and the imprinted genes Z4, Z5, Z6, Z10 and Z11 have an abnormal expression number of the imprinted gene of less than 1.5%. Or the total expression level of the imprinted genes Z4, Z5, Z6, Z10 and Z11 is less than 30% or a combination of at least two;
  • the imprinted gene Z4, Z5, Z6, Z10, and Z11 have an imprinted gene expression level of 16-20%, and the imprinted genes Z4, Z5, Z6, Z10, and Z11 have abnormal expression levels of the imprinted gene copy number. 1.5-2.5% or a total expression amount of the imprinting genes Z4, Z5, Z6, Z10 and Z11 of any one of 30-40% or a combination of at least two;
  • the imprinted gene Z4, Z5, Z6, Z10 and Z11 have an imprinted gene expression level of 20-25%, and the imprinted genes Z4, Z5, Z6, Z10 and Z11 have an abnormal expression level of the imprinted gene copy number.
  • 2.5-4% or a total expression amount of the imprinted genes Z4, Z5, Z6, Z10 and Z11 is any one of 40-50% or a combination of at least two;
  • the imprinted gene Z4, Z5, Z6, Z10 and Z11 have an imprinted gene expression level of 25-30%, and the imprinted genes Z4, Z5, Z6, Z10 and Z11 have an abnormal expression level of the imprinted gene copy number. 4-6% or a total expression amount of the imprinting genes Z4, Z5, Z6, Z10 and Z11 is any one of 50-65% or a combination of at least two;
  • the imprinted gene Z4, Z5, Z6, Z10 and Z11 have an imprinted gene expression level greater than 30%, and the imprinted genes Z4, Z5, Z6, Z10 and Z11 have an abnormal expression number of the imprinted gene greater than 6%. Or the total expression level of the imprinted genes Z4, Z5, Z6, Z10 and Z11 is greater than 65% or a combination of at least two;
  • the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount of the imprinted genes Z4, Z5, Z6, Z10, and Z11 are independent of each other.
  • the five different grades of the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount of Z16 are:
  • any of the imprinted gene Z16 has an imprinted gene deletion expression of less than 10%, the imprinted gene Z16 has an imprinted gene copy number abnormal expression amount of less than 1%, or the imprinted gene Z16 has a total expression amount of less than 25%.
  • the imprinted gene Z16 has an imprinted gene deletion expression of 10-15%, the imprinted gene Z16 has an imprinted gene copy number abnormal expression of 1-2% or the imprinted gene Z16 has a total expression of 25%. Any one of -30% or a combination of at least two;
  • the imprinted gene Z16 has an imprinted gene deletion expression of 15-25%, the imprinted gene Z16 has an imprinted gene copy number abnormal expression of 2-4% or the imprinted gene Z16 has a total expression of 30%. Any one of -40% or a combination of at least two;
  • the imprinted gene Z16 has an imprinted gene deletion expression amount of 25-30%, the imprinted gene Z16 has an imprinted gene copy number abnormal expression amount of 4-6% or the imprinted gene Z16 has a total expression amount of 40%. Any one of -55% or a combination of at least two;
  • Grade IV the imprinting gene deletion expression amount of the imprinting gene Z16 is greater than 30%, the imprinting gene copy number abnormal expression amount of the imprinting gene Z16 is greater than 6%, or the total expression amount of the imprinting gene Z16 is greater than 55% One or a combination of at least two;
  • the five different grades of the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount of Z16 are:
  • the imprinted gene Z16 has an imprinted gene deletion expression amount of less than 15%, the imprinted gene Z16 has an imprinted gene copy number abnormal expression amount of less than 1.5%, or the imprinted gene Z16 has a total expression amount of less than 30%.
  • the imprinted gene Z16 has an imprinted gene deletion expression of 15-20%, the imprinted gene Z16 has an imprinted gene copy number abnormal expression of 1.5-4% or the imprinted gene Z16 has a total expression of 30%. Any one of -40% or a combination of at least two;
  • the imprinted gene Z16 has an imprinted gene deletion expression of 20-25%, the imprinted gene Z16 has an imprinted gene copy number abnormal expression of 4-7% or the imprinted gene Z16 has a total expression of 40%. -50% of any one or a combination of at least two;
  • the imprinted gene Z16 has an imprinted gene deletion expression amount of 25-30%, the imprinted gene Z16 has an imprinted gene copy number abnormal expression amount of 7-10% or the imprinted gene Z16 has a total expression amount of 50%. -60% of any one or a combination of at least two;
  • Grade IV any of the imprinted gene Z16 having an imprinted gene deletion expression amount greater than 30%, the imprinted gene Z16 imprinted gene copy number abnormal expression amount being greater than 10%, or the imprinted gene Z16 total expression amount being greater than 60% One or a combination of at least two.
  • the five different grades of the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount of Z8 are:
  • the imprinted gene Z8 has an imprinted gene deletion expression amount of less than 16%, the imprinted gene Z8 has an imprinted gene copy number abnormal expression amount of less than 2.5%, or the imprinted gene Z8 has a total expression amount of less than 35%.
  • the imprinted gene Z8 has an imprinted gene deletion expression amount of less than 16%, the imprinted gene Z8 has an imprinted gene copy number abnormal expression amount of less than 2.5%, or the imprinted gene Z8 has a total expression amount of less than 35%.
  • the imprinted gene deletion expression amount of the imprinted gene Z8 is 16-20%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z8 is 2.5-4%, or the total expression amount of the imprinted gene Z8 is 35 Any one of -40% or a combination of at least two;
  • the imprinted gene deletion expression amount of the imprinted gene Z8 is 20-25%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z8 is 4-6.5%, or the total expression amount of the imprinted gene Z8 is 40 -50% of any one or a combination of at least two;
  • the imprinted gene Z8 has an imprinted gene deletion expression amount of 25-30%, the imprinted gene Z8 has an imprinted gene copy number abnormal expression amount of 6.5-8% or the imprinted gene Z8 has a total expression amount of 50%. -65% of any one or a combination of at least two;
  • Grade IV the imprinted gene deletion expression amount of the imprinted gene Z8 is greater than 30%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z8 is greater than 8%, or the total expression amount of the imprinted gene Z8 is greater than 65% One or a combination of at least two;
  • the five different grades of the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount of Z8 are:
  • the imprinted gene deletion expression amount of the imprinted gene Z8 is less than 10%, the abnormal expression level of the imprinted gene copy number of the imprinted gene Z8 is less than 1%, or the total expression amount of the imprinted gene Z8 is less than 25%.
  • the imprinted gene deletion expression amount of the imprinted gene Z8 is less than 10%, the abnormal expression level of the imprinted gene copy number of the imprinted gene Z8 is less than 1%, or the total expression amount of the imprinted gene Z8 is less than 25%.
  • the imprinted gene deletion expression amount of the imprinted gene Z8 is 10-15%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z8 is 1-2%, or the total expression amount of the imprinted gene Z8 is 25 Any one of -30% or a combination of at least two;
  • the imprinted gene deletion expression amount of the imprinted gene Z8 is 15-25%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z8 is 2-3%, or the total expression amount of the imprinted gene Z8 is 30 -45% of any one or a combination of at least two;
  • the imprinted gene Z8 has an imprinted gene deletion expression amount of 25-30%, the imprinted gene Z8 has an imprinted gene copy number abnormal expression amount of 3-5% or the imprinted gene Z8 has a total expression amount of 45%. Any one of -55% or a combination of at least two;
  • Grade IV the imprinted gene deletion expression amount of the imprinted gene Z8 is greater than 30%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z8 is greater than 5%, or the total expression amount of the imprinted gene Z8 is greater than 55% One or a combination of at least two.
  • the present disclosure provides a device for detecting the degree of benign and malignant esophageal tumors and/or gastric tumors, which employs the above-described imprinted gene grading model, comprising the following units:
  • sampling unit obtaining a sample to be tested
  • Probe design unit design specific primers according to the imprinted gene sequence
  • the analysis unit calculates the total expression amount of the imprinted gene, the imprinted gene deletion expression amount, and the abnormal expression amount of the imprinted gene copy number, and the imprinted gene deletion expression amount and the imprinted gene copy number abnormality are obtained by the imprinted gene classification model.
  • the level of expression and the total expression level of the imprinted gene are used to determine the degree of benign and malignant esophageal tumors and/or gastric tumors.
  • the imprinted gene is deleted after the cells are subjected to hematoxylin staining, and there are two red/brown labeled nuclei in the nucleus.
  • the abnormal copy number of the imprinted gene is that after the cells are subjected to hematoxylin staining, there are more than two red/brown marks in the nucleus.
  • the copy number abnormality is caused by abnormal gene replication of cancer cells, resulting in the expression of this gene as triploid or even higher polyploid.
  • the hematoxylin-stained label is selected from, but not limited to, red or brown, and staining with other colors can also be used for calculation of total expression of imprinted gene, amount of imprinted gene deletion, and abnormal expression level of imprinted gene copy number.
  • the detection system is used for early and intuitive observation of changes in the imprinted and traced genes of the esophagus and gastric tumors at the cellular and tissue levels to determine the benign and malignant extent of the tumor, and to provide the most favorable treatment for patients with early esophageal and gastric tumors. opportunity.
  • the present disclosure provides a method for detecting the degree of benign and malignant esophageal tumors and/or gastric tumors, using the imprinted gene staging model, comprising the steps of:
  • the analysis unit by calculating the imprinted gene deletion expression amount and the imprinted gene copy number abnormal expression amount, by the imprinted gene grading model according to any one of claims 1 to 9, thereby imprinting the gene deletion expression amount and the imprinted gene
  • the level of abnormal expression of copy number is used to diagnose the degree of benign and malignant esophageal tumors and/or gastric tumors.
  • the sample to be tested described in step (1) is derived from human tissues and/or cells.
  • the sample to be tested is feasible as long as the RNA is subjected to a timely fixation process, and can be selected by a person skilled in the art as needed, and is not particularly limited herein.
  • the sample to be tested in the present disclosure includes paraffin sections of the tissue and esophageal mucosa falling off. Any one or combination of at least two of the cell or gastroscopic biopsy samples.
  • the specific operation procedure of the paraffin section of the tissue is to obtain a human tumor tissue sample, which is fixed in 10% neutral formalin in time, embedded in paraffin, cut into 10 ⁇ m thick, and made into a tissue film with a positively charged slide; Only 10 ⁇ m thick, so some of the microscopically seen nuclei are incomplete, so some false negative gene deletions will occur.
  • the specific operation procedure of the esophageal mucosal exfoliated cells is to obtain mucosal exfoliated cells by esophageal pull net examination, and timely fix with 10% neutral formalin.
  • the specific operation procedure of the gastroscope biopsy sample is to obtain human cells of the esophagus or stomach under the gastroscope, and the 10th neutral formalin can be fixed in time.
  • the sampling process is simple. Compared with the blood circulation characteristics, esophageal mucosal exfoliated cells and gastroscopic biopsy can be located. Esophageal mucosal exfoliated cells and gastroscopic biopsy have special effects as experimental samples. The advantages.
  • the sample to be tested is an esophageal mucosal exfoliated cell and/or a gastroscopic biopsy sample.
  • the imprinting genes are Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16, the imprinting gene Z1 is Gnas, and the imprinting gene Z3 is Peg10, The imprinted gene Z4 is Igf2r, the imprinted gene Z5 is Mest, the imprinted gene Z6 is Plagl1, the imprinted gene Z8 is Dcn, the imprinted gene Z10 is Gatm, the imprinted gene Z11 is Grb10, and the imprinted gene Z13 is Sgce, and the imprinting gene Z16 is Snrpn/Snurf.
  • the imprinted genes Z1 (Gnas), Z3 (Peg10), Z4 (Igf2r), Z5 (Mest), Z6 (Plagl1), Z8 (Dcn), Z10 (Gatm), Z11 (Grb10), Z13 (Sgce), Z16 (Snrpn/Snurf) has different degrees of expression in normal tumor cell tissues, and the expression and imprinting state will change significantly when malignant lesions occur.
  • the design probes are based on the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16, namely Gnas, Peg10, Igf2r, Mest, Plagl, Dcn, Gatm, Grb10, Sgce and Snrpn/ Designed by Snurf, a sequence was selected as a probe within the internal loop of each gene, and the specific probe was designed by Advanced Cell Diagnostics.
  • the in situ hybridization employs an RNAscope in situ hybridization method.
  • the RNAscope in situ hybridization method uses a single or multi-channel colorimetric kit or a single or multi-channel fluorescent kit, preferably a single channel red/brown color kit or multiple channels. Fluorescent kit.
  • the multi-channel coloring kit or the multi-channel fluorescent kit comprises two or more channels of coloring kits or fluorescent kits, and the two-channel coloring kit or multi-channel fluorescent kit can use two The combined expression of imprinted gene probes or imprinted genes and other genes is even the combined expression of multiple imprinted genes and non-imprinted genes.
  • the formula for calculating the total expression amount of the imprinted gene, the amount of the imprinted gene, and the abnormal expression amount of the imprinted gene in the model are as follows:
  • Imprinted gene deletion gene expression level (LOI) c / (b + c + d) ⁇ 100%;
  • a is a cell nucleus in which there is no label in the nucleus and no imprinted gene is expressed after the hematoxylin staining of the cell; and b is a red/brown mark in the nucleus after the hematoxylin staining of the cell, and the imprinting gene exists.
  • the nucleus; the c is a hematoxylin staining of the cells, there are two red/brown marks in the nucleus, and the nucleus of the imprinted gene is deleted; and the d is a hematoxylin staining of the cells, and there are more than two red/brown marks in the nucleus.
  • imprinted gene copy number abnormal cell nucleus is a cell nucleus in which there is no label in the nucleus and no imprinted gene is expressed after the hematoxylin staining of the cell
  • b is a red/brown mark in the nucleus after the hematoxylin staining of the cell,
  • the hematoxylin-stained label is selected from, but not limited to, red or brown, and staining with other colors can also be used for calculation of the total expression level of the imprinted gene, the amount of imprinted gene deletion, and the abnormal expression level of the imprinted gene copy number.
  • the probe was amplified by in situ hybridization, and Hemotoxy (hematoxylin) nuclei were used to amplify the signal, and under the 40 ⁇ or 60 ⁇ microscope, the presence of imprinted genes, imprinted gene deletion or copy number abnormality in each nucleus was determined, and the imprint was calculated.
  • the total gene expression level, the amount of the imprinted gene deletion gene, and the gene expression amount of the imprinted gene copy number abnormality were used to determine the degree of benign and malignant tumors of the sample. Since the section is only 10 microns, about 20% of the nuclei seen under the microscope are incomplete nuclei, which means that there is a possibility of partial false negatives.
  • the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount are divided into five different levels.
  • the five different grades count at least 1200 cells in the region where the probe is most positive for each probe, for ten of Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16
  • the imprinted gene deletion expression amount of the imprinted gene, the abnormal expression level of the imprinted gene copy number, and the total expression amount of the imprinted gene were separately divided.
  • the five different levels of the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount for Z1 are:
  • the imprinted gene deletion expression amount of the imprinted gene Z1 is less than 16%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z1 is less than 2.5%, or the total expression amount of the imprinted gene Z1 is less than 35% One or a combination of at least two;
  • the imprinted gene deletion expression amount of the imprinted gene Z1 is 16-20%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z1 is 2.5-4%, or the total expression amount of the imprinted gene Z1 is 35 Any one of -40% or a combination of at least two;
  • the imprinted gene deletion expression amount of the imprinted gene Z1 is 20-25%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z1 is 4-6.5%, or the total expression amount of the imprinted gene Z1 is 40. -50% of any one or a combination of at least two;
  • the imprinted gene deletion expression amount of the imprinted gene Z1 is 25-30%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z1 is 6.5-8%, or the total expression amount of the imprinted gene Z1 is 50 -65% of any one or a combination of at least two;
  • Grade IV the imprinted gene deletion expression amount of the imprinted gene Z1 is greater than 30%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z1 is greater than 8%, or the total expression amount of the imprinted gene Z1 is greater than 65% One or a combination of at least two.
  • the five different levels of the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount for Z3 and Z13 are:
  • the imprinted gene deletion expression amount of the imprinted genes Z3 and Z13 is less than 10%, the imprinted gene copy number abnormal expression amount of the imprinted genes Z3 and Z13 is less than 1% or the total expression amount of the imprinted genes Z3 and Z13 Any one of less than 25% or a combination of at least two;
  • the imprinted gene Z3 and Z13 have an imprinted gene deletion expression of 10-15%, the imprinted genes Z3 and Z13 have an imprinted gene copy number abnormal expression of 1-2% or the imprinted genes Z3 and Z13
  • the total expression amount is any one of 25-30% or a combination of at least two;
  • the imprinted gene Z3 and Z13 have an imprinted gene deletion expression of 15-25%, the imprinted genes Z3 and Z13 have an imprinted gene copy number abnormal expression of 2-3% or the imprinted genes Z3 and Z13
  • the total expression amount is any one of 30-45% or a combination of at least two;
  • the imprinted gene Z3 and Z13 have an imprinted gene deletion expression of 25-30%, the imprinted genes Z3 and Z13 have an imprinted gene copy number abnormal expression of 3-5% or the imprinted genes Z3 and Z13
  • the total expression amount is any one of 45-55% or a combination of at least two;
  • Grade IV the imprinted gene deletion expression amount of the imprinted genes Z3 and Z13 is greater than 30%, the imprinted gene copy number abnormal expression amount of the imprinted genes Z3 and Z13 is greater than 5% or the total expression amount of the imprinted genes Z3 and Z13 Any one of more than 55% or a combination of at least two;
  • the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount of the imprinted genes Z3 and Z13 are independent of each other.
  • the five different levels of the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount for Z4, Z5, Z6, Z10, and Z11 are:
  • the imprinted gene Z4, Z5, Z6, Z10 and Z11 have an imprinted gene deletion expression of less than 16%, and the imprinted genes Z4, Z5, Z6, Z10 and Z11 have an abnormal expression number of the imprinted gene of less than 1.5%. Or the total expression level of the imprinted genes Z4, Z5, Z6, Z10 and Z11 is less than 30% or a combination of at least two;
  • the imprinted gene Z4, Z5, Z6, Z10, and Z11 have an imprinted gene expression level of 16-20%, and the imprinted genes Z4, Z5, Z6, Z10, and Z11 have abnormal expression levels of the imprinted gene copy number. 1.5-2.5% or a total expression amount of the imprinting genes Z4, Z5, Z6, Z10 and Z11 of any one of 30-40% or a combination of at least two;
  • the imprinted gene Z4, Z5, Z6, Z10 and Z11 have an imprinted gene expression level of 20-25%, and the imprinted genes Z4, Z5, Z6, Z10 and Z11 have an abnormal expression level of the imprinted gene copy number.
  • 2.5-4% or a total expression amount of the imprinted genes Z4, Z5, Z6, Z10 and Z11 is any one of 40-50% or a combination of at least two;
  • the imprinted gene Z4, Z5, Z6, Z10 and Z11 have an imprinted gene expression level of 25-30%, and the imprinted genes Z4, Z5, Z6, Z10 and Z11 have an abnormal expression level of the imprinted gene copy number. 4-6% or a total expression amount of the imprinting genes Z4, Z5, Z6, Z10 and Z11 is any one of 50-65% or a combination of at least two;
  • the imprinted gene Z4, Z5, Z6, Z10 and Z11 have an imprinted gene expression level greater than 30%, and the imprinted genes Z4, Z5, Z6, Z10 and Z11 have an abnormal expression number of the imprinted gene greater than 6%. Or the total expression level of the imprinted genes Z4, Z5, Z6, Z10 and Z11 is greater than 65% or a combination of at least two;
  • the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount of the imprinted genes Z4, Z5, Z6, Z10, and Z11 are independent of each other.
  • the five different grades of the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount of Z16 are:
  • any of the imprinted gene Z16 has an imprinted gene deletion expression of less than 10%, the imprinted gene Z16 has an imprinted gene copy number abnormal expression amount of less than 1%, or the imprinted gene Z16 has a total expression amount of less than 25%.
  • the imprinted gene Z16 has an imprinted gene deletion expression of 10-15%, the imprinted gene Z16 has an imprinted gene copy number abnormal expression of 1-2% or the imprinted gene Z16 has a total expression of 25%. Any one of -30% or a combination of at least two;
  • the imprinted gene Z16 has an imprinted gene deletion expression of 15-25%, the imprinted gene Z16 has an imprinted gene copy number abnormal expression of 2-4% or the imprinted gene Z16 has a total expression of 30%. Any one of -40% or a combination of at least two;
  • the imprinted gene Z16 has an imprinted gene deletion expression amount of 25-30%, the imprinted gene Z16 has an imprinted gene copy number abnormal expression amount of 4-6% or the imprinted gene Z16 has a total expression amount of 40%. Any one of -55% or a combination of at least two;
  • Grade IV the imprinting gene deletion expression amount of the imprinting gene Z16 is greater than 30%, the imprinting gene copy number abnormal expression amount of the imprinting gene Z16 is greater than 6%, or the total expression amount of the imprinting gene Z16 is greater than 55% One or a combination of at least two.
  • the five different grades of the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount of Z16 are:
  • the imprinted gene Z16 has an imprinted gene deletion expression amount of less than 15%, the imprinted gene Z16 has an imprinted gene copy number abnormal expression amount of less than 1.5%, or the imprinted gene Z16 has a total expression amount of less than 30%.
  • the imprinted gene Z16 has an imprinted gene deletion expression of 15-20%, the imprinted gene Z16 has an imprinted gene copy number abnormal expression of 1.5-4% or the imprinted gene Z16 has a total expression of 30%. Any one of -40% or a combination of at least two;
  • the imprinted gene Z16 has an imprinted gene deletion expression of 20-25%, the imprinted gene Z16 has an imprinted gene copy number abnormal expression of 4-7% or the imprinted gene Z16 has a total expression of 40%. -50% of any one or a combination of at least two;
  • the imprinted gene Z16 has an imprinted gene deletion expression amount of 25-30%, the imprinted gene Z16 has an imprinted gene copy number abnormal expression amount of 7-10% or the imprinted gene Z16 has a total expression amount of 50%. -60% of any one or a combination of at least two;
  • Grade IV any of the imprinted gene Z16 having an imprinted gene deletion expression amount greater than 30%, the imprinted gene Z16 imprinted gene copy number abnormal expression amount being greater than 10%, or the imprinted gene Z16 total expression amount being greater than 60% One or a combination of at least two.
  • the five different grades of the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount of Z8 are:
  • the imprinted gene deletion expression amount of the imprinted gene Z8 is less than 16%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z8 is less than 2.5%, or the total expression amount of the imprinted gene Z8 is less than 35% One or a combination of at least two;
  • the imprinted gene deletion expression amount of the imprinted gene Z8 is 16-20%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z8 is 2.5-4%, or the total expression amount of the imprinted gene Z8 is 35 Any one of -40% or a combination of at least two;
  • the imprinted gene deletion expression amount of the imprinted gene Z8 is 20-25%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z8 is 4-6.5%, or the total expression amount of the imprinted gene Z8 is 40 -50% of any one or a combination of at least two;
  • the imprinted gene Z8 has an imprinted gene deletion expression amount of 25-30%, the imprinted gene Z8 has an imprinted gene copy number abnormal expression amount of 6.5-8% or the imprinted gene Z8 has a total expression amount of 50%. -65% of any one or a combination of at least two;
  • Grade IV the imprinted gene deletion expression amount of the imprinted gene Z8 is greater than 30%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z8 is greater than 8%, or the total expression amount of the imprinted gene Z8 is greater than 65% One or a combination of at least two;
  • the five different grades of the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount of Z8 are:
  • the imprinted gene deletion expression amount of the imprinted gene Z8 is less than 10%, the abnormal expression level of the imprinted gene copy number of the imprinted gene Z8 is less than 1%, or the total expression amount of the imprinted gene Z8 is less than 25%.
  • the imprinted gene deletion expression amount of the imprinted gene Z8 is less than 10%, the abnormal expression level of the imprinted gene copy number of the imprinted gene Z8 is less than 1%, or the total expression amount of the imprinted gene Z8 is less than 25%.
  • the imprinted gene deletion expression amount of the imprinted gene Z8 is 10-15%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z8 is 1-2%, or the total expression amount of the imprinted gene Z8 is 25 Any one of -30% or a combination of at least two;
  • the imprinted gene deletion expression amount of the imprinted gene Z8 is 15-25%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z8 is 2-3%, or the total expression amount of the imprinted gene Z8 is 30 -45% of any one or a combination of at least two;
  • the imprinted gene Z8 has an imprinted gene deletion expression amount of 25-30%, the imprinted gene Z8 has an imprinted gene copy number abnormal expression amount of 3-5% or the imprinted gene Z8 has a total expression amount of 45%. Any one of -55% or a combination of at least two;
  • Grade IV the imprinted gene deletion expression amount of the imprinted gene Z8 is greater than 30%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z8 is greater than 5%, or the total expression amount of the imprinted gene Z8 is greater than 55% One or a combination of at least two.
  • the imprinted gene Z16 was uniformly expressed in the stroma and epithelial cells of gastric tumor tissues, but was significantly different in the early stage of gastric cancer before the clinical stage II stage and the advanced stage gastric cancer of the clinical stage III-IV stage.
  • the degree of benign and malignant diagnosis of esophageal tumors is classified into benign tumors, esophageal cancer potential, early esophageal cancer, meta-esophageal cancer, and advanced esophageal cancer.
  • the result of the diagnosis of the degree of benign and malignant esophagus is the imprinted gene deletion expression amount and the imprinted gene copy number of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16.
  • the abnormal expression level was less than grade I, and the imprinted gene deletion level of no more than one imprinted gene in the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 was grade I and the imprinted gene Z1
  • the result of the diagnosis of the degree of benign and malignant esophagus is that the imprinted gene deletion expression level of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is grade I, imprinted
  • the abnormal expression level of the imprinted gene copy number of at least two imprinted genes of genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is grade I or imprinted genes Z1, Z3, Z4, Z5, Z6,
  • the imprinted gene deletion expression of no more than one imprinted gene in Z8, Z10, Z11, Z13 and Z16 is class II and the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16
  • the abnormal expression level of the imprinted gene copy number of more than one imprinted gene is any one of the second grade, and is the esophageal cancer potential;
  • the result of the diagnosis of the degree of benign and malignant esophagus is that the imprinted gene deletion expression level of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is Grade II, imprinted
  • the abnormal expression level of the imprinted gene copy number of at least two imprinted genes of genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is Grade II or imprinted genes Z1, Z3, Z4, Z5, Z6,
  • the imprinted gene deletion expression level of no more than one imprinted gene in Z8, Z10, Z11, Z13 and Z16 is grade III and the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 do not If the abnormal expression level of the imprinted gene copy number of more than one imprinted gene is in any of the grade III, it is early esophageal cancer;
  • the result of the diagnosis of the degree of benign and malignant esophagus is that the imprinted gene deletion expression level of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is grade III, imprinted
  • the abnormal expression level of imprinted gene copy number of at least two imprinted genes of genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is grade III or imprinted genes Z1, Z3, Z4, Z5, Z6,
  • the imprinted gene deletion expression of no more than one imprinted gene in Z8, Z10, Z11, Z13 and Z16 is IV and the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 do not If the abnormal expression level of the imprinted gene copy number of more than one imprinted gene is in any of the IV grades, it is a metaphase esophageal cancer;
  • the result of the diagnosis of the degree of benign and malignant esophagus is that the imprinted gene of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 has a deletion level of IV or imprinted gene.
  • the abnormal expression level of the imprinted gene copy number of at least two of the Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13, Z8, Z10, Z11, Z13 and Z16 is grade IV, and is advanced esophageal cancer.
  • the diagnosis of benign and malignant gastric tumors is classified into benign gastric tumors, gastric cancer potential, early gastric cancer, metaphase gastric cancer, and advanced gastric cancer.
  • the results of the diagnosis of the degree of benign and malignant tumors of the gastric tumors are that the imprinted gene Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 have an imprinted gene expression level and an abnormal expression amount of the imprinted gene copy number are less than I.
  • the imprinted gene of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13, and Z16 has an imprinted gene deletion level of I and the imprinted genes Z1, Z3, Z4, Z5
  • the abnormal expression level of the imprinted gene copy number of not more than one imprinted gene in Z6, Z8, Z10, Z11, Z13, and Z16 is any one of grade I, and is a benign tumor;
  • the result of the diagnosis of the degree of benign and malignant of the gastric tumor is that the imprinted gene of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 has a deletion level of I, and the imprinted gene
  • the abnormal expression level of imprinted gene copy number of at least two imprinted genes of Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 is grade I
  • imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10 is expressed in the class I and has no more than one imprinted gene in the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13.
  • the abnormal expression level of the imprinted gene copy number is grade I and the imprinted gene deletion expression amount of the imprinted gene Z16 or the abnormal expression level of the imprinted gene copy number is grade I or above, or the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10
  • the imprinting gene with no more than one imprinted gene in Z11 and Z13 has a deletion level of II and imprinted with no more than one imprinted gene in the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13.
  • the abnormal expression level of the gene copy number is in any of the cases of grade II, which is the potential of gastric cancer;
  • the result of the diagnosis of the degree of benign and malignant of the gastric tumor is that the imprinted gene of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 has a deletion level of II, imprinted gene
  • the abnormal expression level of the imprinted gene copy number of at least two imprinted genes in Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 is Grade II or imprinted genes Z1, Z3, Z4, Z5, Z6, Z8,
  • the imprinted gene deletion expression level of no more than one imprinted gene in Z10, Z11 and Z13 is grade III and no more than one imprinted gene in the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 If the abnormal expression level of the imprinted gene copy is in any of the grade III, it is early gastric cancer;
  • the result of the diagnosis of the degree of benign and malignant of the gastric tumor is that the imprinted gene of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 has a deletion level of III, imprinted gene
  • the abnormal expression level of imprinted gene copy number of at least 2 imprinted genes of Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 is grade III or imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10
  • the imprinting gene with no more than one imprinted gene in Z11 and Z13 is expressed at the IV level and the imprint of no more than one imprinted gene in the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 If the abnormal expression level of the gene copy number is in any of the IV grades, it is a metaphase gastric cancer;
  • the result of diagnosing the degree of benign and malignant of the gastric tumor is that the imprinted gene of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 has a deletion level of IV or imprinted gene Z1.
  • the abnormal expression level of the imprinted gene copy number of at least two of the Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z6, Z8, Z10, Z11 and Z13 is grade IV, and is advanced gastric cancer.
  • the present disclosure provides the imprinted gene grading model and/or the device for esophageal tumor and gastric tumor detection.
  • the present disclosure provides the use of the imprinted gene grading model and/or the device for the preparation of a medicament or device for treating an esophageal tumor and/or a gastric tumor.
  • the degree of benign and malignant diagnosis of esophageal tumors is classified into benign tumors, esophageal cancer potential, early esophageal cancer, meta-esophageal cancer, and advanced esophageal cancer.
  • the results of the diagnosis of the degree of benign and malignant tumors of the esophageal tumor are that the imprinted gene Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 have an imprinted gene expression level and an abnormal expression amount of the imprinted gene copy number are less than I.
  • the imprinted gene of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13, and Z16 has an imprinted gene deletion level of I and the imprinted genes Z1, Z3, Z4, Z5
  • the abnormal expression level of the imprinted gene copy number of not more than one imprinted gene in Z6, Z8, Z10, Z11, Z13, and Z16 is any one of grade I, and is a benign tumor;
  • the result of the diagnosis of the degree of benign and malignant of the esophageal tumor is that the imprinted gene expression level of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is Grade I,
  • the abnormal expression level of the imprinted gene copy number of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is Grade I or imprinted genes Z1, Z3, Z4, Z5, Z6 In the Z8, Z10, Z11, Z13 and Z16, the imprinted gene deletion expression level of the imprinted gene is Grade II and the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16
  • the abnormal expression level of the imprinted gene copy number of no more than one imprinted gene is any one of the second grade, and is the esophageal cancer potential;
  • the result of the diagnosis of the degree of benign and malignant of the esophageal tumor is that the imprinted gene deletion expression level of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is II,
  • the abnormal expression level of the imprinted gene copy number of at least two imprinted genes of the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is Grade II or imprinted genes Z1, Z3, Z4, Z5, Z6 In the Z8, Z10, Z11, Z13 and Z16, the imprinted gene deletion expression level of the imprinted gene is III and the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 If the abnormal expression level of the imprinted gene copy number of no more than one imprinted gene is in any of the grade III, it is early esophageal cancer;
  • the result of the diagnosis of the degree of benign and malignant of the esophageal tumor is that the imprinted gene deletion expression level of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is III,
  • the abnormal expression level of imprinted gene copy number of at least two imprinted genes of imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is grade III or imprinted genes Z1, Z3, Z4, Z5, Z6 , the imprinting gene of Z8, Z10, Z11, Z13 and Z16 is not more than one imprinted gene, and the imprinted gene is expressed in the IV level and in the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16. If the abnormal expression level of the imprinted gene copy number of no more than one imprinted gene is any of the IV grades, it is a metaphase esophageal cancer;
  • the result of the diagnosis of the degree of benign and malignant tumors of the esophageal tumor is that the imprinted gene of at least two of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 has an imprinted gene expression level of IV or imprinted.
  • the abnormal expression level of the imprinted gene copy number of at least two of the imprinted genes in the genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13, and Z16 is Grade IV, and is an advanced esophageal cancer.
  • the diagnosis of gastric tumors is classified into benign gastric tumors, gastric cancer potential, early gastric cancer, metaphase gastric cancer, and advanced gastric cancer.
  • the results of the diagnosis of the degree of benign and malignant tumors of the gastric tumors are that the imprinted gene Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 have an imprinted gene expression level and an abnormal expression amount of the imprinted gene copy number are less than I.
  • the imprinted gene of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13, and Z16 has an imprinted gene deletion level of I and the imprinted genes Z1, Z3, Z4, Z5
  • the abnormal expression level of the imprinted gene copy number of not more than one imprinted gene in Z6, Z8, Z10, Z11, Z13, and Z16 is any one of grade I, and is a benign tumor;
  • the result of the diagnosis of the degree of benign and malignant of the gastric tumor is that the imprinted gene of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 has a deletion level of I, and the imprinted gene
  • the abnormal expression level of imprinted gene copy number of at least two imprinted genes of Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 is grade I
  • imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10 is expressed in the class I and has no more than one imprinted gene in the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13.
  • the abnormal expression level of the imprinted gene copy number is grade I and the imprinted gene deletion expression amount of the imprinted gene Z16 or the abnormal expression level of the imprinted gene copy number is grade I or above, or the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10
  • the imprinting gene with no more than one imprinted gene in Z11 and Z13 has a deletion level of II and imprinted with no more than one imprinted gene in the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13.
  • the abnormal expression level of the gene copy number is in any of the cases of grade II, which is the potential of gastric cancer;
  • the result of the diagnosis of the degree of benign and malignant of the gastric tumor is that the imprinted gene of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 has a deletion level of II, imprinted gene
  • the abnormal expression level of the imprinted gene copy number of at least two imprinted genes in Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 is Grade II or imprinted genes Z1, Z3, Z4, Z5, Z6, Z8,
  • the imprinted gene deletion expression level of no more than one imprinted gene in Z10, Z11 and Z13 is grade III and no more than one imprinted gene in the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 If the abnormal expression level of the imprinted gene copy is in any of the grade III, it is early gastric cancer;
  • the result of the diagnosis of the degree of benign and malignant of the gastric tumor is that the imprinted gene of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 has a deletion level of III, imprinted gene
  • the abnormal expression level of imprinted gene copy number of at least 2 imprinted genes of Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 is grade III or imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10
  • the imprinting gene with no more than one imprinted gene in Z11 and Z13 is expressed at the IV level and the imprint of no more than one imprinted gene in the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 If the abnormal expression level of the gene copy number is in any of the IV grades, it is a metaphase gastric cancer;
  • the result of diagnosing the degree of benign and malignant of the gastric tumor is that the imprinted gene of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 has a deletion level of IV or imprinted gene Z1.
  • the abnormal expression level of the imprinted gene copy number of at least two of the Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z6, Z8, Z10, Z11 and Z13 is grade IV, and is advanced gastric cancer.
  • the present disclosure provides a marker for the diagnosis of advanced gastric cancer, the marker being the imprinting gene Z1 and the imprinting gene Z11;
  • imprinting gene Z1 is Gnas
  • imprinting gene Z11 is Grb10
  • the advanced gastric cancer is gastric cancer having a TNM rating of gastric cancer greater than T2N0M0.
  • the imprinted gene Z16 was uniformly expressed in the stroma and epithelial cells of gastric tumor tissues, but was significantly different in the early stage of gastric cancer before the clinical stage II stage and the advanced stage gastric cancer of the clinical stage III-IV stage. In early gastric cancer, especially in the period of intraepithelial neoplasia, 75% of the imprinted gene Z16 has an imprinted deletion, copy number abnormality and total expression level of more than grade I. With the deterioration of cancer, 90% of advanced gastric cancer samples The imprinted gene Z16 has an imprinted deletion, an abnormal copy number, and a total expression level of 0.
  • the imprinted gene Z16 combined with the imprinting genes Z1 and Z11 is a good diagnostic marker for the diagnosis of early gastric cancer, especially the transition from low-grade intraepithelial neoplasia to gastric cancer, but the imprinting gene Z16 is useful in advanced gastric cancer. Not big, only need Z1 and Z11 to make a diagnosis.
  • the detection model and the device are used to visually express the performance of the imprinted gene on the sample of the esophageal tumor and/or the gastric tumor patient, and the method for in situ labeling the imprinted gene Objective, intuitive, early, accurate detection of changes in the imprinted (trace) gene, and can provide quantitative models that make a significant contribution to the diagnosis of molecular pathology.
  • 1 is a pathological section of gastric cancer of the present disclosure by hematoxylin-stained nuclei, wherein the a is a hematoxylin-stained cell, and there is no label in the nucleus, and the imprinted gene is not expressed; the b is a cell nucleus after hematoxylin staining There is a red/brown mark in the presence of the imprinted gene; the c is the hematoxylin staining of the cells, there are two red/brown marks in the nucleus, and the imprinted gene is deleted; the d is the cell nucleus memory after hematoxylin staining In more than two red/brown markers, the imprinted gene copy number is abnormal;
  • Fig. 2(a) shows the expression status of 10 genes in the pathological section of grade 0 gastric tumor
  • Fig. 2(b) shows the expression status of 10 genes in the pathological section of grade I gastric cancer
  • Fig. 2(c) shows grade II gastric cancer.
  • the expression status of 10 genes in the pathological section Fig. 2(d) shows the expression status of 10 genes in the pathological section of grade III gastric cancer
  • Fig. 2(e) shows the expression status of 10 genes in the pathological section of grade IV gastric cancer. ;
  • Fig. 3(a) shows the intensity of imprinting deletion of gastric cancer genes Z1, Z11 and Z16
  • Fig. 3(b) shows the intensity of copy number abnormalities of gastric cancer Z1, Z11 and Z16 for gastric cancer
  • Fig. 3(c) is an imprint.
  • Fig. 3(d) are the intensities of the imprinted genes Z3, Z4, Z5, Z6, Z8, Z10 and Z13 on the gastric cancer
  • Fig. 3(e) is The intensity of the imprinting genes Z3, Z4, Z5, Z6, Z8, Z10 and Z13 on the copy number abnormality of gastric cancer
  • Fig. 3(e) is The intensity of the imprinting genes Z3, Z4, Z5, Z6, Z8, Z10 and Z13 on the copy number abnormality of gastric cancer
  • 3(f) is the total expression of the imprinted genes Z3, Z4, Z5, Z6, Z8, Z10 and Z13 on gastric cancer.
  • the intensity of the quantity wherein LOI is the expression level of the imprinted gene deletion gene, CNV is the gene expression quantity of the imprinted gene copy number abnormality, and TE is the total expression level of the imprinted gene;
  • Fig. 4(a) shows the intensity of the imprinting gene Z1 imprinting deletion, copy number abnormality and total expression amount
  • Fig. 4(b) shows the intensity of the imprinting gene Z11 imprinting deletion, copy number abnormality and total expression amount
  • FIG. 4(c) is The imprinting gene Z16 imprinted deletion, copy number abnormality and total expression level intensity
  • Figure 4 (d) is the imprinting gene Z3 imprint deletion, copy number abnormality and total expression level intensity
  • Figure 4 (e) is the imprinted gene Z4 imprint missing
  • Figure 4 (f) is the imprinting gene Z5 imprint deletion, copy number abnormality and total expression level intensity
  • Figure 4 (g) is imprinted gene Z6 imprint deletion, copy number abnormality and total expression
  • Figure 4 (h) is the intensity of the imprinting gene Z8 imprint deletion, copy number abnormality and total expression
  • Figure 4 (i) is the imprinting gene Z10 imprint deletion, copy number abnormality and total expression intensity
  • Figure 4 (j) The intensity of imprinting gene Z13 imprinting deletion, copy number abnormality and total expression amount, wherein LOI is the expression level of imprinted gene deletion gene, CNV is the gene expression amount of imprinted
  • Fig. 5(a) shows the distribution range and grading standard of imprinted gene Z1 in 37 cases of gastric tumor pathological sections, imprinting deletion and copy number abnormality
  • Fig. 5(b) is the imprinting gene Z11 applied to 37 cases of gastric tumor pathological section.
  • Fig. 5(c) shows the distribution range and grading standard of imprinting deletion and copy number abnormality in the pathological section of 37 gastric tumors with imprinting gene Z16
  • Fig. 5(d) The imprinted gene Z3 was applied to the pathological sections of 37 cases of gastric tumors, the distribution range and grading standard of imprinting deletion and copy number abnormality
  • Fig. 5(e) was the imprinted gene Z4 applied to 37 cases of gastric tumor pathological sections, imprinted deletion and The distribution range and grading standard of copy number abnormality
  • Fig. 5(f) shows the distribution range and grading standard of imprinting deletion and copy number abnormality in 37 pathological sections of gastric tumors
  • Fig. 5(g) is imprinted gene.
  • Z6 was applied to the pathological sections of 37 cases of gastric tumors, the distribution range and grading standard of imprinting deletion and copy number abnormality.
  • Figure 5(h) shows that imprinted gene Z8 was applied to 37 cases of gastric tumor pathological sections with imprinting deletion and copy number abnormality.
  • Figure 5 (i) is the distribution range and grading standard of imprinting gene Z10 applied to 37 gastric tumor pathological sections, imprinting deletion and copy number abnormality
  • Fig. 5 (j) is applied to imprinting gene Z13 in 37 cases In the pathological section of gastric tumors, the distribution range and grading criteria of imprinting deletion and copy number abnormality, wherein LOI is the expression level of imprinted gene deletion gene, CNV is the gene expression quantity of imprinted gene copy number abnormality, and TE is the total expression level of imprinted gene;
  • Fig. 6(a) shows the expression status of 10 genes in the pathological section of grade 0 esophageal tumor
  • Fig. 6(b) shows the expression status of 10 genes in the pathological section of grade I esophageal cancer
  • Fig. 6(c) is level II.
  • Fig. 6(d) shows the expression status of 10 genes in the pathological section of grade III esophageal cancer
  • Fig. 6(e) shows 10 cases in the pathological section of grade IV esophageal cancer.
  • Fig. 7(a) shows the intensity of imprinting defects in esophageal cancer by imprinting genes Z1, Z11 and Z16
  • Fig. 7(b) shows the intensity of copying abnormalities of esophageal cancer by imprinting genes Z1, Z11 and Z16
  • Fig. 7(c) For the intensity of the total expression of the imprinted genes Z1, Z11 and Z16 for esophageal cancer
  • Fig. 7(d) shows the intensity of imprinting defects of the imprinted genes Z3, Z4, Z5, Z6, Z8, Z10 and Z13 for esophageal cancer
  • Fig. 7(d) shows the intensity of imprinting defects of the imprinted genes Z3, Z4, Z5, Z6, Z8, Z10 and Z13 for esophageal cancer
  • Figure 8 (a) shows the intensity of the imprinted gene Z1 imprint deletion, copy number abnormality and total expression
  • Figure 8 (b) shows the imprinted gene Z11 imprint deletion, copy number abnormality and total expression intensity
  • Figure 8 (c) For the imprinting gene Z16 imprint deletion, copy number abnormality and total expression intensity
  • Figure 8 (d) is the imprinting gene Z3 imprint deletion, copy number abnormality and total expression intensity
  • Figure 8 (e) is the imprinted gene Z4 imprint deletion , copy number abnormality and total expression intensity
  • Figure 8 (f) is the imprinting gene Z5 imprint deletion, copy number abnormality and total expression intensity
  • Figure 8 (g) is the imprinted gene Z6 imprint deletion, copy number abnormality and total The intensity of the expression amount
  • 8(h) shows the intensity of the imprinted gene Z8 imprint deletion, copy number abnormality and total expression amount
  • Fig. 8(i) shows the intensity of the imprinted gene Z10 imprint deletion, copy number abnormality and total expression amount
  • 8(j) is the intensity of imprinting gene Z13 imprinting deletion, copy number abnormality and total expression amount, wherein LOI is the expression level of imprinted gene deletion gene, CNV is the gene expression amount of imprinted gene copy number abnormality, and TE is imprinted gene total expression. the amount;
  • Fig. 9(a) shows the distribution range and grading standard of imprinting deletion and copy number abnormality in 57 cases of esophageal tumor pathological sections
  • Fig. 9(b) is the imprinting gene Z11 applied to 57 esophage tumor pathological sections.
  • Fig. 9(c) shows the distribution range and grading standard of imprinting deletion and copy number abnormality in 57 pathological sections of esophageal tumor, which is imprinted gene Z16
  • Fig. 9(b) shows the distribution range and grading standard of imprinting deletion and copy number abnormality in 57 cases of esophageal tumor pathological sections
  • Fig. 9(b) is the imprinting gene Z11 applied to 57 esophage tumor pathological sections.
  • Fig. 9(c) shows the distribution range and grading standard of imprinting deletion and copy number abnormality in 57 pathological sections of esophageal tumor, which is imprinted gene Z16
  • the imprinting gene Z3 was applied to 57 cases of esophageal tumor pathological sections, the distribution range and grading standard of imprinting deletion and copy number abnormality
  • Fig. 9(e) was the imprinting gene Z4 applied to 57 esophage tumor pathological sections, imprinting deletion and The distribution range and grading standard of copy number abnormality
  • Fig. 9(f) shows the distribution range and grading standard of imprinting deletion and copy number abnormality in 57 pathological sections of esophageal tumor
  • the imprinting gene is shown in Fig.
  • FIG. 9(g) Z6 was applied to the pathological sections of esophageal tumors in 57 cases, and the distribution range and grading standard of imprinting deletion and copy number abnormality.
  • Figure 9(h) shows that imprinted gene Z8 was applied to 57 cases of esophageal tumor pathological sections, and the imprint was missing.
  • the distribution range and grading standard of copy number abnormality Fig. 9(i) shows the distribution range and grading standard of imprinting deletion and copy number abnormality in 57 pathological sections of esophageal tumor
  • Fig. 9(j) is the imprint Gene Z13 was applied to the pathological sections of esophageal tumors, and the distribution range and grading criteria of imprinting deletion and copy number abnormality.
  • LOI was the expression level of imprinted gene deletion gene
  • CNV was the gene expression level of imprinted gene copy number abnormality
  • TE was The total expression level of the imprinted gene.
  • Genomic imprinting is a way of gene regulation in epigenetics. It is characterized by methylation of alleles from a particular parent, such that one gene has only one allele and the other is in a state of gene silencing. This type of gene is called a blot (remember) gene. Deletion of the blot is an epigenetic change in which the allelic gene of the imprinted gene results in a silenced allele being activated and beginning to express the gene. Numerous studies have shown that this phenomenon (missing of blots) is ubiquitous in various types of cancer and occurs earlier than cell and tissue morphological changes. At the same time, in healthy cells, the proportion of imprinted deletions is extremely low, in sharp contrast to cancer cells. Therefore, the methylation status of the imprinted gene can be used as a pathological marker to analyze the abnormal state of the cell by a specific molecular detection technique.
  • the detection model and system of the present disclosure express the impression of the imprinted defect on the sample of the esophageal and gastric tumor patients in an intuitive manner, and objectively, intuitively, earlyly and accurately detect the imprint by the method of in situ labeling of the imprinted gene. (trace) changes in genes and can provide quantitative models that make a significant contribution to the diagnosis of esophageal and gastric tumors;
  • the detection device of the present invention can determine the degree of benign and malignant esophageal and gastric tumors and the extent of invasion of cancer cells by gastroscopic biopsy samples before operation in patients with esophagus and stomach tumors, thereby providing a basis for surgery and precise treatment, which is a diagnosis in the field of cell molecules.
  • the present disclosure can accurately determine the grades of esophageal and gastric tumors, and clearly classify the early lesions of esophageal and gastric tumors, ie, the degree of malignancy of intraepithelial neoplasia, by combination of imprinted genes, especially for early and advanced gastric cancer.
  • Accurate differentiation has greatly improved the early and clear diagnosis of esophageal cancer and gastric cancer, especially in early census and postoperative follow-up of cancer, especially for follow-up of suspected relapsed patients, which can win time and save lives for patients.
  • Major contribution
  • the disclosed detection method is different from the immunohistochemical method, and reduces false positives and other negative effects, and not only the targeted drugs which are silenced, eliminated, and rearranged by the discovery of the missing regions of the esophageal and gastric tumor-associated imprinted genes. Or technical methods that can be used to guide later treatment and medication.
  • the method for detecting the imprinted gene comprises the following steps:
  • tissue cell sections (10 ⁇ m) of gastric tumors and fixing them in a 10% neutral formalin solution to prevent RNA degradation for 24 hours, paraffin embedding (FFPE), The slide needs to be loaded with a positive charge, and the slice is baked in an oven at 40 ° C for more than 3 hours;
  • FFPE paraffin embedding
  • Design probe design specific primers according to the imprinted gene sequence
  • the design probe is based on the imprinting genes Z1 (Gnas), Z3 (Peg10), Z4 (Igf2r), Z5 (Mest), Z6 (Plagl1), Z8 (Dcn), Z10 (Gatm), Z11 (Grb10), Z13. (Sgce) and Z16 (Snrpn/Snurf) were designed to select a sequence within the internal loop of each gene as a probe. The specific probe was designed by Advanced Cell Diagnostics.
  • the formula for calculating the total expression of the imprinted gene, the amount of the imprinted gene, and the abnormal expression of the imprinted gene in the model are as follows:
  • Imprinted gene deletion gene expression level (LOI) c / (b + c + d) ⁇ 100%;
  • a, b, c, and d are as shown in FIG. 1 , wherein a is a cell nucleus in which no hemoglobin is stained in the nucleus and the imprinted gene is not expressed; and b is a hematoxylin staining of the cell.
  • the c is the hematoxylin staining of the cell, there are two red/brown marks in the nucleus, and the nucleus of the imprinted gene is deleted;
  • the d is the hematoxylin After staining, there are more than two red/brown markers in the nucleus, and the nuclei with abnormal copy number of the imprinted gene are imprinted.
  • the gastroscopic biopsy sample was obtained by taking a suspicious diseased tissue under a gastroscope and fixing the 10% neutral formalin solution for more than 24 hours.
  • the other detection methods were the same as those in Example 1.
  • each imprinted gene Z1 has an imprinted deletion and an increased expression level.
  • the stage of malignant potential begins to appear, and it rises rapidly to a very high level in the development of gastric cancer from early to late stage.
  • the abnormal copy number of imprinted gene Z1 begins to appear in the malignant potential stage, rises rapidly in early and middle stage gastric cancer, and rises in advanced gastric cancer stage.
  • the speed is slowed down; the imprinting of the imprinted gene Z11 begins to appear in the malignant potential stage, rapidly rises in the early stage of gastric cancer, and the rate of increase in the metastatic gastric cancer slows down, and continues to rise to a higher level in the advanced stage of gastric cancer, and the copy number of the imprinted gene Z11 is abnormal.
  • the increase of expression and expression began to appear in the malignant potential stage, and gradually increased to a higher level in the development of early and late gastric cancer; the imprinting deletion, copy number abnormality and expression increase of the imprinted gene Z16 increased rapidly in the malignant potential and early gastric cancer stage. However, there was a significant decrease in the stage of advanced gastric cancer;
  • the imprinting deletion, copy number abnormality and expression amount of the imprinted gene Z3 appear in the early stage of gastric cancer, continue to rise in the metaphase gastric cancer, and increase in the advanced gastric cancer.
  • the abnormal copy number of the imprinted gene Z4 rises rapidly in the early stage of gastric cancer, in the stage of intermediate and advanced gastric cancer.
  • the rate of increase of the immunization gene Z4 began to appear in the malignant potential stage, and it increased rapidly in the early stage of gastric cancer.
  • the stage of gastric cancer did not increase significantly, and it continued to rise in the advanced stage of gastric cancer.
  • the imprinting of the imprinted gene Z5 was in the stage of early gastric cancer. Rapidly rising, the rate of increase in the mid-stage gastric cancer phase slowed down, and the stage of advanced gastric cancer continued to rise to a higher level.
  • the abnormal copy number of the imprinted gene Z5 began to appear in the early stage of gastric cancer, rapidly increased in the metaphase gastric cancer, and increased in the advanced gastric cancer stage.
  • the expression of the imprinted gene Z5 is increased in the early stomach
  • the stage began to appear, gradually rising to a higher level in the metaphase and advanced gastric cancer; the imprinting of the imprinted gene Z6 began to appear in the early stage of gastric cancer, continued to rise in the metaphase gastric cancer, and the rate of increase in the advanced gastric cancer stage slowed down, a copy of the imprinted gene Z6
  • the number of abnormalities began to appear in the early stage of gastric cancer, and the rate of increase in the metastatic gastric cancer slowed down, and the stage of advanced gastric cancer continued to rise.
  • imprinted gene Z6 began to appear in the early stage of gastric cancer, and slowly increased in the middle and late stage gastric cancer; imprinted genes Z8 imprinting loss and expression increase began to appear in early gastric cancer, and slowly increased in metaphase gastric cancer. The rate of increase in advanced gastric cancer increased, but the level was still not high. The abnormal copy number of imprinted gene Z8 increased rapidly in early gastric cancer.
  • the imprinting deletion, copy number abnormality and expression increase of the imprinted gene Z10 began to appear in the malignant potential stage, and gradually increased to a higher level in the development of early to late gastric cancer; imprinted gene Z13's imprinted deletion and increased expression
  • the stage of malignant potential begins to appear, gradually rising in early and mid-stage gastric cancer, and the rate of increase in late gastric cancer is slowed down.
  • the copy number abnormality of imprinting gene Z13 begins to appear in the malignant potential stage, and the rate of rise in early gastric cancer slows down, and the rate of mid-stage gastric cancer rises. Increased, the rate of increase in advanced gastric cancer slowed down.
  • the tissues of 37 patients with gastric tumors including the gastroscopic biopsy samples (10 micrometers) were obtained in the same manner as in Example 1.
  • any one of the imprinted gene deletion expression amount is less than 16%, the imprinted gene copy number abnormal expression amount is less than 2.5%, or the total imprinted gene expression amount is less than 35%.
  • the combination of at least two is class I, the imprinting gene deletion expression amount is 20-25%, the imprinted gene copy number abnormal expression amount is 4-6.5%, or the imprinted gene total expression amount is 40-50%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 6.5-8%, or the imprinted gene total expression amount is 50-65%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than 8%, or the imprinted gene total expression amount is greater than 65%, or a combination of at least two is IV;
  • any one of the imprinted gene deletion expression amount is less than 16%, the imprinted gene copy number abnormal expression amount is less than 1.5%, or the total imprinted gene expression amount is less than 30%.
  • a combination of at least two of which is level 0 an imprinted gene deletion expression amount of 16-20%, an imprinted gene copy number abnormal expression amount of 1.5-2.5%, or an imprinted gene total expression amount of 30-40% or The combination of at least two is class I, the imprinting gene deletion expression amount is 20-25%, the imprinted gene copy number abnormal expression amount is 2.5-4%, or the imprinted gene total expression amount is 40-50%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 4-6%, or the imprinted gene total expression amount is 50-65%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than
  • the imprinting gene deletion expression amount is less than 15%, the imprinted gene copy number abnormal expression amount is less than 1.5%, or the imprinted gene total expression amount is less than 30%.
  • any one of the imprinted gene deletion expression amount is less than 10%, the imprinted gene copy number abnormal expression amount is less than 1%, or the total imprinted gene expression amount is less than 25%.
  • a combination of at least two of which is 0, an imprinting gene deletion expression amount of 10-15%, an imprinted gene copy number abnormal expression amount of 1-2%, or an imprinted gene total expression amount of 25-30% or
  • the combination of at least two is class I, the imprinting gene deletion expression amount is 15-25%, the imprinted gene copy number abnormal expression amount is 2-3%, or the imprinted gene total expression amount is 30-45%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 3-5%, or the imprinted gene total expression amount is 45-55%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than 5%, or the imprinted
  • the imprinting gene deletion expression amount is less than 16%, the imprinted gene copy number abnormal expression amount is less than 1.5%, or the imprinted gene total expression amount is less than 30%.
  • a combination of at least two of which is level 0 an imprinted gene deletion expression amount of 16-20%, an imprinted gene copy number abnormal expression amount of 1.5-2.5%, or an imprinted gene total expression amount of 30-40% or The combination of at least two is class I, the imprinting gene deletion expression amount is 20-25%, the imprinted gene copy number abnormal expression amount is 2.5-4%, or the imprinted gene total expression amount is 40-50%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 4-6%, or the imprinted gene total expression amount is 50-65%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than 6%
  • any one of the imprinted gene deletion expression amount is less than 16%, the imprinted gene copy number abnormal expression amount is less than 1.5%, or the total imprinted gene expression amount is less than 30%.
  • a combination of at least two of which is level 0 an imprinted gene deletion expression amount of 16-20%, an imprinted gene copy number abnormal expression amount of 1.5-2.5%, or an imprinted gene total expression amount of 30-40% or The combination of at least two is class I, the imprinting gene deletion expression amount is 20-25%, the imprinted gene copy number abnormal expression amount is 2.5-4%, or the imprinted gene total expression amount is 40-50%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 4-6%, or the imprinted gene total expression amount is 50-65%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than
  • any one of the imprinted gene deletion expression amount is less than 16%, the imprinted gene copy number abnormal expression amount is less than 1.5%, or the total imprinted gene expression amount is less than 30%.
  • a combination of at least two of which is level 0 an imprinted gene deletion expression amount of 16-20%, an imprinted gene copy number abnormal expression amount of 1.5-2.5%, or an imprinted gene total expression amount of 30-40% or The combination of at least two is class I, the imprinting gene deletion expression amount is 20-25%, the imprinted gene copy number abnormal expression amount is 2.5-4%, or the imprinted gene total expression amount is 40-50%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 4-6%, or the imprinted gene total expression amount is 50-65%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than
  • any one of the imprinted gene deletion expression amount is less than 10%, the imprinted gene copy number abnormal expression amount is less than 1%, or the total imprinted gene expression amount is less than 25%.
  • the combination of at least two is class I, the imprinting gene deletion expression amount is 15-25%, the imprinted gene copy number abnormal expression amount is 2-3%, or the imprinted gene total expression amount is 30-45%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 3-5%, or the imprinted gene total expression amount is 45-55%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than 5%, or the imprinted
  • any one of the imprinted gene deletion expression amount is less than 16%, the imprinted gene copy number abnormal expression amount is less than 1.5%, or the total imprinted gene expression amount is less than 30%.
  • the combination of at least two is class I, the imprinting gene deletion expression amount is 20-25%, the imprinted gene copy number abnormal expression amount is 2.5-4%, or the imprinted gene total expression amount is 40-50%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 4-6%, or the imprinted gene total expression amount is 50-65%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than 6%, or the imprinted gene total expression amount is greater than 65%, or a combination of at least two is IV;
  • any one of the imprinted gene deletion expression amount is less than 10%, the imprinted gene copy number abnormal expression amount is less than 1%, or the total imprinted gene expression amount is less than 25%.
  • a combination of at least two of which is 0, an imprinting gene deletion expression amount of 10-15%, an imprinted gene copy number abnormal expression amount of 1-2%, or an imprinted gene total expression amount of 25-30% or
  • the combination of at least two is class I, the imprinting gene deletion expression amount is 15-25%, the imprinted gene copy number abnormal expression amount is 2-3%, or the imprinted gene total expression amount is 30-45%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 3-5%, or the imprinted gene total expression amount is 45-55%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than 5%, or the im
  • the degree of benign and malignant diagnosis of tumors is divided into benign tumors, gastric cancer potential, early gastric cancer, metaphase gastric cancer and advanced gastric cancer:
  • the results of the diagnosis of the degree of benign and malignant tumors of the gastric tumors are that the imprinted gene Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 have an imprinted gene expression level and an abnormal expression amount of the imprinted gene copy number are less than I.
  • the imprinted gene of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13, and Z16 has an imprinted gene deletion level of I and the imprinted genes Z1, Z3, Z4, Z5
  • the abnormal expression level of the imprinted gene copy number of not more than one imprinted gene in Z6, Z8, Z10, Z11, Z13, and Z16 is any one of grade I, and is a benign tumor;
  • the result of the diagnosis of the degree of benign and malignant of the gastric tumor is that the imprinted gene of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 has a deletion level of I, and the imprinted gene
  • the abnormal expression level of imprinted gene copy number of at least two imprinted genes of Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 is grade I
  • imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10 is expressed in the class I and has no more than one imprinting gene in the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13.
  • the abnormal expression level of the imprinted gene copy number is grade I and the imprinted gene deletion expression amount of the imprinted gene Z16 or the abnormal expression level of the imprinted gene copy number is grade I or above, or the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10
  • the imprinting gene with no more than one imprinted gene in Z11 and Z13 has a deletion level of II and imprinted with no more than one imprinted gene in the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13.
  • the abnormal expression level of the gene copy number is in any of the cases of grade II, which is the potential of gastric cancer;
  • the result of the diagnosis of the degree of benign and malignant of the gastric tumor is that the imprinted gene of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 has a deletion level of II, imprinted gene
  • the abnormal expression level of the imprinted gene copy number of at least two imprinted genes in Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 is Grade II or imprinted genes Z1, Z3, Z4, Z5, Z6, Z8,
  • the imprinted gene deletion expression level of no more than one imprinted gene in Z10, Z11 and Z13 is grade III and no more than one imprinted gene in the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 If the abnormal expression level of the imprinted gene copy is in any of the grade III, it is early gastric cancer;
  • the result of the diagnosis of the degree of benign and malignant of the gastric tumor is that the imprinted gene of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 has a deletion level of III, imprinted gene
  • the abnormal expression level of imprinted gene copy number of at least 2 imprinted genes of Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 is grade III or imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10
  • the imprinting gene with no more than one imprinted gene in Z11 and Z13 is expressed at the IV level and the imprint of no more than one imprinted gene in the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 If the abnormal expression level of the gene copy number is in any of the IV grades, it is a metaphase gastric cancer;
  • the result of diagnosing the degree of benign and malignant of the gastric tumor is that the imprinted gene of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 has a deletion level of IV or imprinted gene Z1.
  • the abnormal expression level of the imprinted gene copy number of at least two of the Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z6, Z8, Z10, Z11 and Z13 is grade IV, and is advanced gastric cancer.
  • Tissue cell sections (10 micrometers) of esophageal tumors were obtained, and the detection method was the same as in Example 1.
  • the gastroscopic biopsy sample of the esophagus is a suspicious diseased tissue in which the esophagus is taken out under a gastroscope, and the 10% neutral formalin solution is fixed for more than 24 hours.
  • the other detection methods are the same as those in the first embodiment.
  • Fig. 8(a)-Fig. 8(j) The sensitivity of each imprinted gene to esophageal cancer is shown in Fig. 8(a)-Fig. 8(j). It can be seen from Fig. 8(a)-Fig. 8(c) that the imprinting of the imprinted gene Z1 is in the stage of malignant potential. It began to appear, and did not increase significantly in early esophageal cancer. It gradually increased to a high level in the metaphase and advanced esophageal cancer. The copy number abnormality of the imprinted gene Z1 gradually increased from the malignant potential to the advanced esophageal cancer stage, and the expression level of the imprinted gene Z1 was increased.
  • the increase is slowly rising in the development of esophageal cancer; the imprinting of the imprinted gene Z11 begins to appear in the malignant potential stage, and does not increase significantly in the early and middle esophageal cancer stages, and there is a significant increase in the late stage of esophageal cancer, a copy of the imprinted gene Z11
  • the number of abnormalities increased rapidly during the malignant potential stage, and the rate of increase in the development of early and late esophageal cancer gradually slowed down.
  • the increase in the expression of the imprinted gene Z11 did not increase significantly from malignant potential to metaphase esophageal cancer, but in advanced esophageal cancer.
  • the expression of the imprinted gene Z8 increased in the malignant potential stage, but did not increase significantly during the development of esophageal cancer; the imprinted imprinted gene Z10 was deleted in the middle esophageal cancer. The stage began to appear, and did not continue to rise in the advanced esophageal cancer stage. The abnormal copy number of the imprinted gene Z10 began to appear in the metaphase esophageal cancer stage, and continued to rise in advanced esophageal cancer, but the level was not high, and the expression level of the imprinted gene Z10 increased in malignancy.
  • the latent stage began to appear, but did not increase significantly during the development of esophageal cancer; the imprinted deletion of the imprinted gene Z13 began to appear in early esophageal cancer, gradually increased in the metaphase and advanced esophageal cancer stages, but the level was not high, and the imprinted gene Z13 was copied.
  • the number of abnormalities and increased expression increased rapidly in the mid-stage esophageal cancer stage and slowed down in the advanced esophageal cancer stage.
  • the gut biopsy samples (10 micrometers) of the esophagus were obtained from 57 patients with esophageal tumors.
  • the detection method was the same as in Example 1.
  • any one of the imprinted gene deletion expression amount is less than 16%, the imprinted gene copy number abnormal expression amount is less than 2.5%, or the total imprinted gene expression amount is less than 35%.
  • the combination of at least two is class I, the imprinting gene deletion expression amount is 20-25%, the imprinted gene copy number abnormal expression amount is 4-6.5%, or the imprinted gene total expression amount is 40-50%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 6.5-8%, or the imprinted gene total expression amount is 50-65%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than 8%, or the imprinted gene total expression amount is greater than 65%, or a combination of at least two is IV;
  • any one of the imprinted gene deletion expression amount is less than 16%, the imprinted gene copy number abnormal expression amount is less than 1.5%, or the total imprinted gene expression amount is less than 30%.
  • a combination of at least two of which is level 0 an imprinted gene deletion expression amount of 16-20%, an imprinted gene copy number abnormal expression amount of 1.5-2.5%, or an imprinted gene total expression amount of 30-40% or The combination of at least two is class I, the imprinting gene deletion expression amount is 20-25%, the imprinted gene copy number abnormal expression amount is 2.5-4%, or the imprinted gene total expression amount is 40-50%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 4-6%, or the imprinted gene total expression amount is 50-65%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than
  • any one of the imprinted gene deletion expression amount is less than 10%, the imprinted gene copy number abnormal expression amount is less than 1%, or the total imprinted gene expression amount is less than 25%.
  • a combination of at least two of which is 0, an imprinting gene deletion expression amount of 10-15%, an imprinted gene copy number abnormal expression amount of 1-2%, or an imprinted gene total expression amount of 25-30% or
  • the combination of at least two is class I, the imprinting gene deletion expression amount is 15-25%, the imprinted gene copy number abnormal expression amount is 2-4%, or the imprinted gene total expression amount is 30-40%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 4-6%, or the imprinted gene total expression amount is 40-55%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than 6%, or the
  • any one of the imprinted gene deletion expression amount is less than 10%, the imprinted gene copy number abnormal expression amount is less than 1%, or the total imprinted gene expression amount is less than 25%.
  • the combination of at least two is class I, the imprinting gene deletion expression amount is 15-25%, the imprinted gene copy number abnormal expression amount is 2-3%, or the imprinted gene total expression amount is 30-45%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 3-5%, or the imprinted gene total expression amount is 45-55%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than 5%, or the im
  • any one of the imprinted gene deletion expression amount is less than 16%, the imprinted gene copy number abnormal expression amount is less than 1.5%, or the total imprinted gene expression amount is less than 30%.
  • a combination of at least two of which is level 0 an imprinted gene deletion expression amount of 16-20%, an imprinted gene copy number abnormal expression amount of 1.5-2.5%, or an imprinted gene total expression amount of 30-40% or The combination of at least two is class I, the imprinting gene deletion expression amount is 20-25%, the imprinted gene copy number abnormal expression amount is 2.5-4%, or the imprinted gene total expression amount is 40-50%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 4-6%, or the imprinted gene total expression amount is 50-65%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than
  • any one of the imprinted gene deletion expression amount is less than 16%, the imprinted gene copy number abnormal expression amount is less than 1.5%, or the total imprinted gene expression amount is less than 30%.
  • a combination of at least two of which is level 0 an imprinted gene deletion expression amount of 16-20%, an imprinted gene copy number abnormal expression amount of 1.5-2.5%, or an imprinted gene total expression amount of 30-40% or The combination of at least two is class I, the imprinting gene deletion expression amount is 20-25%, the imprinted gene copy number abnormal expression amount is 2.5-4%, or the imprinted gene total expression amount is 40-50%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 4-6%, or the imprinted gene total expression amount is 50-65%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than
  • any one of the imprinted gene deletion expression amount is less than 16%, the imprinted gene copy number abnormal expression amount is less than 1.5%, or the total imprinted gene expression amount is less than 30%.
  • a combination of at least two of which is level 0 an imprinted gene deletion expression amount of 16-20%, an imprinted gene copy number abnormal expression amount of 1.5-2.5%, or an imprinted gene total expression amount of 30-40% or The combination of at least two is class I, the imprinting gene deletion expression amount is 20-25%, the imprinted gene copy number abnormal expression amount is 2.5-4%, or the imprinted gene total expression amount is 40-50%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 4-6%, or the imprinted gene total expression amount is 50-65%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than
  • the imprinting gene deletion expression amount is less than 16%, the imprinted gene copy number abnormal expression amount is less than 2.5%, or the imprinted gene total expression amount is less than 35%.
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than
  • any one of the imprinted gene deletion expression amount is less than 16%, the imprinted gene copy number abnormal expression amount is less than 1.5%, or the total imprinted gene expression amount is less than 30%.
  • the combination of at least two is class I, the imprinting gene deletion expression amount is 20-25%, the imprinted gene copy number abnormal expression amount is 2.5-4%, or the imprinted gene total expression amount is 40-50%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 4-6%, or the imprinted gene total expression amount is 50-65%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than 6%, or the imprinted gene total expression amount is greater than 65%, or a combination of at least two is IV;
  • any one of the imprinted gene deletion expression amount is less than 10%, the imprinted gene copy number abnormal expression amount is less than 1%, or the total imprinted gene expression amount is less than 25%.
  • the combination of at least two is class I, the imprinting gene deletion expression amount is 15-25%, the imprinted gene copy number abnormal expression amount is 2-3%, or the imprinted gene total expression amount is 30-45%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 3-5%, or the imprinted gene total expression amount is 45-55%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than 5%, or the
  • Diagnosis of the degree of benign and malignant esophageal tumors can be divided into benign tumors, esophageal cancer potential, early esophageal cancer, meta-esophageal cancer and advanced esophageal cancer:
  • the results of the diagnosis of the degree of benign and malignant tumors of the esophageal tumor are that the imprinted gene Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 have an imprinted gene expression level and an abnormal expression amount of the imprinted gene copy number are less than I.
  • the imprinted gene of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13, and Z16 has an imprinted gene deletion level of I and the imprinted genes Z1, Z3, Z4, Z5
  • the abnormal expression level of the imprinted gene copy number of not more than one imprinted gene in Z6, Z8, Z10, Z11, Z13, and Z16 is any one of grade I, and is a benign tumor;
  • the result of the diagnosis of the degree of benign and malignant of the esophageal tumor is that the imprinted gene expression level of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is Grade I,
  • the abnormal expression level of the imprinted gene copy number of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is Grade I or imprinted genes Z1, Z3, Z4, Z5, Z6 In the Z8, Z10, Z11, Z13 and Z16, the imprinted gene deletion expression level of the imprinted gene is Grade II and the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16
  • the abnormal expression level of the imprinted gene copy number of no more than one imprinted gene is any one of the second grade, and is the esophageal cancer potential;
  • the result of the diagnosis of the degree of benign and malignant of the esophageal tumor is that the imprinted gene deletion expression level of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is II,
  • the abnormal expression level of the imprinted gene copy number of at least two imprinted genes of the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is Grade II or imprinted genes Z1, Z3, Z4, Z5, Z6 In the Z8, Z10, Z11, Z13 and Z16, the imprinted gene deletion expression level of the imprinted gene is III and the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 If the abnormal expression level of the imprinted gene copy number of no more than one imprinted gene is in any of the grade III, it is early esophageal cancer;
  • the result of the diagnosis of the degree of benign and malignant of the esophageal tumor is that the imprinted gene deletion expression level of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is III,
  • the abnormal expression level of imprinted gene copy number of at least two imprinted genes of imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is grade III or imprinted genes Z1, Z3, Z4, Z5, Z6 , the imprinting gene of Z8, Z10, Z11, Z13 and Z16 is not more than one imprinted gene, and the imprinted gene is expressed in the IV level and in the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16. If the abnormal expression level of the imprinted gene copy number of no more than one imprinted gene is any of the IV grades, it is a metaphase esophageal cancer;
  • the result of the diagnosis of the degree of benign and malignant tumors of the esophageal tumor is that the imprinted gene of at least two of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 has an imprinted gene expression level of IV or imprinted.
  • the abnormal expression level of the imprinted gene copy number of at least two of the imprinted genes in the genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13, and Z16 is Grade IV, and is an advanced esophageal cancer.
  • the detection model and system of the present disclosure express the impression of the imprinted defect on the sample of the esophageal and gastric tumor patients in an intuitive manner, and the method of in situ labeling of the imprinted gene is objective, intuitive, early, Accurate detection of changes in the imprinted (trace) gene and the ability to provide a quantitative model that contributes significantly to the diagnosis of esophageal and gastric tumors.

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Abstract

一种用于检测食道肿瘤和/或胃肿瘤良恶性程度的分级模型及其应用，所述模型通过计算印记基因缺失表达量、印记基因拷贝数异常表达量和印记基因总表达量对印记基因在肿瘤中的变化进行分级，所述模型以直观的方法表现了印记缺失在食道肿瘤和胃肿瘤病人的组织和细胞样本上的表现，通过对印记基因原位标记的方法，客观、直观、早期、精确地检测出印记（迹）基因的变化，并可以提供量化的模型，为食道肿瘤和胃肿瘤的诊断做出贡献。

Description

一种用于检测食道肿瘤和/或胃肿瘤良恶性程度的分级模型及其应用 技术领域
本公开涉及生物技术领域，涉及基因诊断领域，例如基因诊断领域，例如一种分级模型及其应用，例如一种用于检测食道肿瘤和/或胃肿瘤良恶性程度的分级模型及其应用，例如一组印记基因在检测食道肿瘤和/或胃肿瘤良恶性程度中的分级模型及其组成的装置和应用。
背景技术
胃癌和食道癌是两种常见的消化道肿瘤，2012年全世界新增胃癌病例95.2万，死亡72.3万，新增食道癌病例45.6万，死亡40万。我国是胃癌和食道癌的高发地区，每年胃癌和食道癌的新发病例和死亡人数都占世界的50％左右。早期胃癌和食道癌的5年生存率超过90％，而中晚期胃癌和食道癌的5年生存率不到30％，因此胃癌和食道癌的早期诊断对拯救患者生命具有非常重要的意义。但是胃癌和食道癌在早期通常没有明显的症状，在我国的检出率低于10％，超过一半的患者在发现胃癌或食道癌时已经是中晚期。
目前已经公认上皮内瘤变是胃的癌前病变，根据增生的程度可以分为低级别上皮内瘤变和高级别上皮内瘤变。低级别上皮内瘤变发展为胃癌的可能性约为20％，而高级别上皮内瘤变则有超过70％的可能会发展为胃癌。目前病理学上对低级别和高级别上皮内瘤变的区分主要由病理医生根据形态学进行主观判断。虽然已经有多项研究尝试使用粘液组织化学、免疫细胞化学检测分化或细胞增殖标记物，以及癌基因产物和形态测量分析来消除分类的主观性，但至今还没有比传统的形态学分析更具有明显的优势。类似的，目前食道癌的诊断也主要依赖于影像学和组织形态学，诊断的准确率在很大程度上依赖于医生的经 验和主观判断，容易造成误诊。
CN 106609037 A公开了一种基于多个基因诊断胃癌患者的检测试剂盒，所述试剂盒中包括了诊断胃癌的引物，可以用于诊断胃癌，但其检测不够精准，不能够区分胃癌处于具体哪个阶段。
传统病理学对细胞的良恶性诊断是基于细胞的大小，形态，浸润性和周边细胞组织的关系来作出判断的。它对细胞(癌症)的早期变化的发现有很大的局限性，因此细胞分子水平的癌症诊断方法，一度成为研究热点。随着人们在分子生物学领域的不断深入研究，越来越多的分子检测技术被运用到癌症诊断中。
癌症的产生是随时间推移而累积的表观遗传改变和基因上的变异所导致的不受控制的细胞生长/分裂。传统病理学诊断根据细胞和组织的大小，形态和结构上的变异，从而做出食道和胃肿瘤良恶性判断。随着分子生物学的发展与深入，越来越多的分子检测技术被应用于食道癌和胃癌的检测。从癌症的发展过程分析，分子层面的改变(表观遗传学和基因学)远早于细胞形态和组织结构的变异。所以分子生物学检测对癌症早期的检测更敏感。
基于上述原因，目前的食道癌和胃癌诊断需要新的检测系统和检测模型，基于患者活检样本，解析食道癌和胃癌在细胞层面上存在的分子标记物变化，以此提供更精确的预诊和诊断信息。
发明内容
针对现有技术的不足及实际的需求，本公开提供了一种印记基因分级模型及其应用。
为达到上述目的，本公开采用以下技术方案：
第一方面，本公开提供了一种用于食道肿瘤和/或胃肿瘤的印记基因分级模 型，所述模型通过计算印记基因缺失表达量、印记基因拷贝数异常表达量和印记基因的总表达量在食道肿瘤和/或胃肿瘤中的变化对印记基因的表达状态进行分级；
其中，所述印记基因为Z1、Z11或Z16中的任意一个或至少两个的组合，所述印记基因Z1为Gnas，所述印记基因Z11为Grb10，所述印记基因Z16为Snrpn/Snurf。
所述印记(迹)缺失是指印记(迹)基因中原先处在沉寂状态的等位基因被激活(去甲基化)，是癌症中最常见和最早期就发生的表观遗传改变，并且这个特性可以用作病理标记。相对而言，在健康细胞检测中，印迹缺失比例很低，所述印记基因与印迹基因同时一个概念，表示同一个意思，可以进行替换。
公开人发现通过计算Z1、Z11和Z16中任意一个印记基因在食道肿瘤和/或胃肿瘤中的印记基因缺失表达量和印记基因拷贝数异常表达量，对食道癌的诊断敏感度可以达到90.4％以上，对胃癌的诊断敏感度可以达到67.6％以上。
在本文一实施例中，若初步检测只检测一个印记基因，可以检测Z1、Z11和Z16中的任意一个印记基因。
在本文一实施例中，用于检测食道癌时最优选的印记基因为Z1。
在本文一实施例中，用于检测胃癌时最优选的印记基因为Z11。
在本文一实施例中，发明人发现，单独检测一个Z1印记基因，对食道癌的诊断敏感度可以达到96.3％，单独检测一个Z11印记基因，对食道癌的诊断敏感度可以达到92.5％，单独检测一个Z16印记基因，对食道癌的诊断敏感度可以达到90.4％。
在本文一实施例中，发明人发现，单独检测一个Z1印记基因，对胃癌的诊断敏感度可以达到80.6％，单独检测一个Z11印记基因，对胃癌的诊断敏感度可 以达到88.9％，单独检测一个Z16印记基因，对胃癌的诊断敏感度可以达到67.6％。
在本文一实施例中，若检测印记基因的两个印记基因的组合，所述组合可以是Z1、Z11和Z16中的任意两个，例如所述组合可以是Z1和Z11的组合，Z1和Z16的组合或Z11和Z16的组合。
发明人发现通过计算两个或两个以上的印记基因的印记基因缺失表达量和印记基因拷贝数异常表达量可以进一步提高敏感度，检测印记基因的两个印记基因的组合，对食道癌的诊断敏感度可以达到98.1％以上，检测Z1和Z16的组合时，对食道癌的诊断敏感度可以达到98.1％，检测Z1和Z11的组合时，对食道癌的诊断敏感度可以达到99.2％，检测Z11和Z16的组合时，对食道癌的诊断敏感度可以达到99.2％；
发明人发现通过计算两个或两个以上的印记基因的印记基因缺失表达量和印记基因拷贝数异常表达量可以进一步提高敏感度，检测印记基因的两个印记基因的组合，对胃癌的诊断敏感度可以达到94.1％以上，检测Z1和Z16的组合时，对胃癌的诊断敏感度可以达到97.1％，检测Z1和Z11的组合时，对胃癌的诊断敏感度可以达到97.2％，检测Z11和Z16的组合时，对胃癌的诊断敏感度可以达到94.1％。
在本文一实施例中，所述印记基因还包括Z3、Z4、Z5、Z6、Z8、Z10或Z13中的任意一个或至少两个的组合；其中，所述印记基因Z3为Peg10，所述印记基因Z4为Igf2r，所述印记基因Z5为Mest，所述印记基因Z6为Plagl1，所述印记基因Z8为Dcn，所述印记基因Z10为Gatm，所述印记基因Z13为Sgce。
本中，人发现在使用Z1、Z11和Z16基因检测的基础上再增加Z3、Z4、Z5、Z6、Z8、Z10、Z13基因进行联合诊断，不仅有助于增加检测的准确度，而且增 加其他探针辅助诊断可以进一步避免假阳性的出现，能够将检测准确度进一步提高，从而能够实现所有食道和胃肿瘤样本的精确分级和判断。
在本文一实施例中，所述模型计算印记基因的方法为：计算印记基因的组合，所述印记基因为Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16的十个基因的组合。
所述印记基因缺失为将细胞进行苏木素染色后，细胞核内存在两个红色/棕色标记，所述印记基因拷贝数异常为将细胞进行苏木素染色后，细胞核内存在两个以上红色/棕色标记，所述拷贝数异常是由于癌细胞异常地进行基因复制，导致这个基因表达时呈现为三倍体甚至更高的多倍体的情况。
所述苏木素染色后的标记选自但不限于红色或棕色，用其他颜色进行染色标记也可用于印迹基因总表达量、印记基因缺失表达量和印记基因拷贝数异常表达量的计算。
在本文一实施例中，所述计算印记基因总表达量、印记基因缺失表达量和印记基因拷贝数异常表达量的公式如下：
总表达量＝(b+c+d)/(a+b+c+d)×100％；
正常印记基因总表达量＝b/(b+c+d)×100％；
印记基因缺失基因表达量(LOI)＝c/(b+c+d)×100％；
印记基因拷贝数异常的基因表达量(CNV)＝d/(b+c+d)×100％；
其中，所述a为将细胞进行苏木素染色后，细胞核内不存在标记，印记基因没有表达的细胞核；所述b为将细胞进行苏木素染色后，细胞核内存在一个红色/棕色标记，印记基因存在的细胞核；所述c为将细胞进行苏木素染色后，细胞核内存在两个红色/棕色标记，印记基因缺失的细胞核；所述d为将细胞进行苏木素染色后，细胞核内存在两个以上红色/棕色标记，印记基因拷贝数异常 的细胞核。
在本文一实施例中，所述苏木素染色后的标记选自但不限于红色或棕色，用其他颜色进行染色标记也可用于印记基因缺失表达量、印记基因拷贝数异常表达量和印记基因总表达量的计算。
在本文一实施例中，将探针通过原位杂交，和Hemotoxy(苏木精)细胞核染色扩增信号，在40×或60×显微镜下，判断每一个细胞核内印记基因存在、印记基因缺失或拷贝数异常，通过计算印记基因缺失基因表达量、印记基因拷贝数异常表达量和印记基因总表达量来判定该样本的肿瘤良恶性程度；由于切片仅为10μm，所以在显微镜下所见细胞核大约有20％为不完整细胞核，也就是说有部分假阴性的可能性存在。
在本文一实施例中，所述印记基因缺失表达量、印记基因拷贝数异常表达量和印记基因总表达量分成五个不同的等级。
在本文一实施例中，通过每个探针在样本表达最阳性的区域对至少1200个细胞进行计数，针对Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16的十个印记基因的印记基因缺失表达量、印记基因拷贝数异常表达量和印记基因总表达量分别进行划分的五个不同的等级。
在本文一实施例中，所述针对Z1的印记基因缺失表达量、印记基因拷贝数异常表达量和总表达量划分的五个不同的等级为：
0级：所述印记基因Z1的印记基因缺失表达量小于16％、所述印记基因Z1的印记基因拷贝数异常表达量小于2.5％或所述印记基因Z1的总表达量小于35％中的任意一种或至少两种的组合；
I级：所述印记基因Z1的印记基因缺失表达量为16-20％、所述印记基因Z1的印记基因拷贝数异常表达量为2.5-4％或所述印记基因Z1的总表达量为 35-40％中的任意一种或至少两种的组合；
II级：所述印记基因Z1的印记基因缺失表达量为20-25％、所述印记基因Z1的印记基因拷贝数异常表达量为4-6.5％或所述印记基因Z1的总表达量为40-50％中的任意一种或至少两种的组合；
III级：所述印记基因Z1的印记基因缺失表达量为25-30％、所述印记基因Z1的印记基因拷贝数异常表达量为6.5-8％或所述印记基因Z1的总表达量为50-65％中的任意一种或至少两种的组合；
IV级：所述印记基因Z1的印记基因缺失表达量大于30％、所述印记基因Z1的印记基因拷贝数异常表达量大于8％或所述印记基因Z1的总表达量大于65％中的任意一种或至少两种的组合。
所述针对Z3和Z13的印记基因缺失表达量、印记基因拷贝数异常表达量和总表达量划分的五个不同的等级为：
0级：所述印记基因Z3和Z13的印记基因缺失表达量小于10％、所述印记基因Z3和Z13的印记基因拷贝数异常表达量小于1％或所述印记基因Z3和Z13的总表达量小于25％中的任意一种或至少两种的组合；
I级：所述印记基因Z3和Z13的印记基因缺失表达量为10-15％、所述印记基因Z3和Z13的印记基因拷贝数异常表达量为1-2％或所述印记基因Z3和Z13的总表达量为25-30％中的任意一种或至少两种的组合；
II级：所述印记基因Z3和Z13的印记基因缺失表达量为15-25％、所述印记基因Z3和Z13的印记基因拷贝数异常表达量为2-3％或所述印记基因Z3和Z13的总表达量为30-45％中的任意一种或至少两种的组合；
III级：所述印记基因Z3和Z13的印记基因缺失表达量为25-30％、所述印记基因Z3和Z13的印记基因拷贝数异常表达量为3-5％或所述印记基因Z3和 Z13的总表达量为45-55％中的任意一种或至少两种的组合；
IV级：所述印记基因Z3和Z13的印记基因缺失表达量大于30％、所述印记基因Z3和Z13的印记基因拷贝数异常表达量大于5％或所述印记基因Z3和Z13的总表达量大于55％中的任意一种或至少两种的组合；
所述印记基因Z3和Z13的印记基因缺失表达量、印记基因拷贝数异常表达量和总表达量是相互独立的。
所述针对Z4、Z5、Z6、Z10和Z11的印记基因缺失表达量、印记基因拷贝数异常表达量和总表达量划分的五个不同的等级为：
0级：所述印记基因Z4、Z5、Z6、Z10和Z11的印记基因缺失表达量小于16％、所述印记基因Z4、Z5、Z6、Z10和Z11的印记基因拷贝数异常表达量小于1.5％或所述印记基因Z4、Z5、Z6、Z10和Z11的总表达量小于30％中的任意一种或至少两种的组合；
I级：所述印记基因Z4、Z5、Z6、Z10和Z11的印记基因缺失表达量为16-20％、所述印记基因Z4、Z5、Z6、Z10和Z11的印记基因拷贝数异常表达量为1.5-2.5％或所述印记基因Z4、Z5、Z6、Z10和Z11的总表达量为30-40％中的任意一种或至少两种的组合；
II级：所述印记基因Z4、Z5、Z6、Z10和Z11的印记基因缺失表达量为20-25％、所述印记基因Z4、Z5、Z6、Z10和Z11的印记基因拷贝数异常表达量为2.5-4％或所述印记基因Z4、Z5、Z6、Z10和Z11的总表达量为40-50％中的任意一种或至少两种的组合；
III级：所述印记基因Z4、Z5、Z6、Z10和Z11的印记基因缺失表达量为25-30％、所述印记基因Z4、Z5、Z6、Z10和Z11的印记基因拷贝数异常表达量为4-6％或所述印记基因Z4、Z5、Z6、Z10和Z11的总表达量为50-65％中的任 意一种或至少两种的组合；
IV级：所述印记基因Z4、Z5、Z6、Z10和Z11的印记基因缺失表达量大于30％、所述印记基因Z4、Z5、Z6、Z10和Z11的印记基因拷贝数异常表达量大于6％或所述印记基因Z4、Z5、Z6、Z10和Z11的总表达量大于65％中的任意一种或至少两种的组合；
所述印记基因Z4、Z5、Z6、Z10和Z11的印记基因缺失表达量、印记基因拷贝数异常表达量和总表达量是相互独立的。
用于检测食道肿瘤时，所述针对Z16的印记基因缺失表达量、印记基因拷贝数异常表达量和总表达量划分的五个不同的等级为：
0级：所述印记基因Z16的印记基因缺失表达量小于10％、所述印记基因Z16的印记基因拷贝数异常表达量小于1％或所述印记基因Z16的总表达量小于25％中的任意一种或至少两种的组合；
I级：所述印记基因Z16的印记基因缺失表达量为10-15％、所述印记基因Z16的印记基因拷贝数异常表达量为1-2％或所述印记基因Z16的总表达量为25-30％中的任意一种或至少两种的组合；
II级：所述印记基因Z16的印记基因缺失表达量为15-25％、所述印记基因Z16的印记基因拷贝数异常表达量为2-4％或所述印记基因Z16的总表达量为30-40％中的任意一种或至少两种的组合；
III级：所述印记基因Z16的印记基因缺失表达量为25-30％、所述印记基因Z16的印记基因拷贝数异常表达量为4-6％或所述印记基因Z16的总表达量为40-55％中的任意一种或至少两种的组合；
IV级：所述印记基因Z16的印记基因缺失表达量大于30％、所述印记基因Z16的印记基因拷贝数异常表达量大于6％或所述印记基因Z16的总表达量大于 55％中的任意一种或至少两种的组合；
用于检测胃肿瘤时，所述针对Z16的印记基因缺失表达量、印记基因拷贝数异常表达量和总表达量划分的五个不同的等级为：
0级：所述印记基因Z16的印记基因缺失表达量小于15％、所述印记基因Z16的印记基因拷贝数异常表达量小于1.5％或所述印记基因Z16的总表达量小于30％中的任意一种或至少两种的组合；
I级：所述印记基因Z16的印记基因缺失表达量为15-20％、所述印记基因Z16的印记基因拷贝数异常表达量为1.5-4％或所述印记基因Z16的总表达量为30-40％中的任意一种或至少两种的组合；
II级：所述印记基因Z16的印记基因缺失表达量为20-25％、所述印记基因Z16的印记基因拷贝数异常表达量为4-7％或所述印记基因Z16的总表达量为40-50％中的任意一种或至少两种的组合；
III级：所述印记基因Z16的印记基因缺失表达量为25-30％、所述印记基因Z16的印记基因拷贝数异常表达量为7-10％或所述印记基因Z16的总表达量为50-60％中的任意一种或至少两种的组合；
IV级：所述印记基因Z16的印记基因缺失表达量大于30％、所述印记基因Z16的印记基因拷贝数异常表达量大于10％或所述印记基因Z16的总表达量大于60％中的任意一种或至少两种的组合。
由于印记基因Z16在胃癌中表达的特殊性，Z16的III级和IV级大部分出现在胃癌的早中期，90％的晚期胃癌样本Z16表达都较低。
用于检测食道肿瘤时，所述针对Z8的印记基因缺失表达量、印记基因拷贝数异常表达量和总表达量划分的五个不同的等级为：
0级：所述印记基因Z8的印记基因缺失表达量小于16％、所述印记基因Z8 的印记基因拷贝数异常表达量小于2.5％或所述印记基因Z8的总表达量小于35％中的任意一种或至少两种的组合；
I级：所述印记基因Z8的印记基因缺失表达量为16-20％、所述印记基因Z8的印记基因拷贝数异常表达量为2.5-4％或所述印记基因Z8的总表达量为35-40％中的任意一种或至少两种的组合；
II级：所述印记基因Z8的印记基因缺失表达量为20-25％、所述印记基因Z8的印记基因拷贝数异常表达量为4-6.5％或所述印记基因Z8的总表达量为40-50％中的任意一种或至少两种的组合；
III级：所述印记基因Z8的印记基因缺失表达量为25-30％、所述印记基因Z8的印记基因拷贝数异常表达量为6.5-8％或所述印记基因Z8的总表达量为50-65％中的任意一种或至少两种的组合；
IV级：所述印记基因Z8的印记基因缺失表达量大于30％、所述印记基因Z8的印记基因拷贝数异常表达量大于8％或所述印记基因Z8的总表达量大于65％中的任意一种或至少两种的组合；
用于检测胃肿瘤时，所述针对Z8的印记基因缺失表达量、印记基因拷贝数异常表达量和总表达量划分的五个不同的等级为：
0级：所述印记基因Z8的印记基因缺失表达量小于10％、所述印记基因Z8的印记基因拷贝数异常表达量小于1％或所述印记基因Z8的总表达量小于25％中的任意一种或至少两种的组合；
I级：所述印记基因Z8的印记基因缺失表达量为10-15％、所述印记基因Z8的印记基因拷贝数异常表达量为1-2％或所述印记基因Z8的总表达量为25-30％中的任意一种或至少两种的组合；
II级：所述印记基因Z8的印记基因缺失表达量为15-25％、所述印记基因 Z8的印记基因拷贝数异常表达量为2-3％或所述印记基因Z8的总表达量为30-45％中的任意一种或至少两种的组合；
III级：所述印记基因Z8的印记基因缺失表达量为25-30％、所述印记基因Z8的印记基因拷贝数异常表达量为3-5％或所述印记基因Z8的总表达量为45-55％中的任意一种或至少两种的组合；
IV级：所述印记基因Z8的印记基因缺失表达量大于30％、所述印记基因Z8的印记基因拷贝数异常表达量大于5％或所述印记基因Z8的总表达量大于55％中的任意一种或至少两种的组合。
发明人发现印记基因Z8和Z16比较特殊，其在食道肿瘤或胃肿瘤中的印记基因缺失表达量、印记基因拷贝数异常表达量和总表达量的分级情况是不同的，所以将印记基因Z8和Z16按照食道肿瘤或胃肿瘤两种情况分开来分级。
在本文一实施例中，本公开提供了一种用于检测食道肿瘤和/或胃肿瘤良恶性程度的装置，其采用上述印记基因分级模型，包括如下单元：
(1)取样单元：获取待测样本；
(2)探针设计单元：根据印记基因序列设计特异性引物；
(3)检测单元：将步骤(2)的探针与待测样本进行原位杂交；
(4)分析单元：显微镜成像分析印记基因的表达情况；
其中，所述分析单元通过计算印记基因总表达量、印记基因缺失表达量和印记基因拷贝数异常表达量，通过所述的印记基因分级模型，从而通过印记基因缺失表达量、印记基因拷贝数异常表达量和印记基因总表达量的等级来判断食道肿瘤和/或胃肿瘤的良恶性程度。
所述印记基因缺失为将细胞进行苏木素染色后，细胞核内存在两个红色/棕色标记的细胞核，所述印记基因拷贝数异常为将细胞进行苏木素染色后，细胞 核内存在两个以上红色/棕色标记的细胞核，所述拷贝数异常是由于癌细胞异常地进行基因复制，导致这个基因表达时呈现为三倍体甚至更高的多倍体的情况。
所述苏木素染色后的标记选自但不限于红色或棕色，用其他颜色进行染色标记也可用于印记基因总表达量、印记基因缺失表达量和印记基因拷贝数异常表达量的计算。
所述检测系统是用于细胞和组织水平下早期直观地观察食道和胃肿瘤的印记(迹)基因的变化从而判断肿瘤的良恶性及恶性程度，为早期食道和胃肿瘤患者提供最有利的治疗机会。
在本文一实施例中，本公开提供一种用于检测食道肿瘤和/或胃肿瘤良恶性程度的方法，采用所述的印记基因分级模型，包括如下步骤：
(1)获取待测样本；
(2)根据印记基因序列设计特异性引物；
(3)将步骤(2)的探针与待测样本进行原位杂交；
(4)显微镜成像分析印记基因的表达情况，从而诊断乳腺肿瘤的良恶性程度；
其中，所述分析单元通过计算印记基因缺失表达量和印记基因拷贝数异常表达量，通过权利要求1-9中任一项所述的印记基因分级模型，从而通过印记基因缺失表达量和印记基因拷贝数异常表达量的等级来诊断食道肿瘤和/或胃肿瘤的良恶性程度。
在本文一实施例中，步骤(1)所述的待测样本来自于人的组织和/或细胞。
所述待测样本只要RNA经过及时固定的处理都是可行的，本领域技术人员可以根据需要进行选择，在此不做特殊限定，本公开所述待测样本包括组织的 石蜡切片、食道黏膜脱落细胞或胃镜活检样本中的任意一种或至少两种的组合。
所述组织的石蜡切片具体操作步骤为获取人体肿瘤组织样本，及时用10％中性福尔马林固定，石蜡包埋，切成10μm厚，用带正电荷的玻片制成组织片子；因为只有10μm厚，因此显微镜下看见的有一部分为不完整的细胞核，所以会出现部分假阴性的基因缺失。
所述食道黏膜脱落细胞具体操作步骤为通过食道拉网检查获取黏膜脱落细胞，及时用10％中性福尔马林固定即可。
所述胃镜活检样本具体操作步骤为在胃镜下获取食道或胃的人体细胞，及时用10％中性福尔马林固定即可。
由于食道黏膜脱落细胞获取原理与胃镜活检样本类似，其样本检测的操作方式与胃镜活检样本类似，其结果也类似。
由于食道黏膜脱落细胞检查和胃镜活检对病人伤害小，取样过程简单，相较于血液的循环特性，食道黏膜脱落细胞和胃镜活检还能定位，食道黏膜脱落细胞和胃镜活检作为实验样本有其特殊的优势。
在本文一实施例中，所述待测样本为食道黏膜脱落细胞和/或胃镜活检样本。
在本文一实施例中，所述印记基因为Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16，所述印记基因Z1为Gnas，所述印记基因Z3为Peg10，所述印记基因Z4为Igf2r，所述印记基因Z5为Mest，所述印记基因Z6为Plagl1，所述印记基因Z8为Dcn，所述印记基因Z10为Gatm，所述印记基因Z11为Grb10，所述印记基因Z13为Sgce，所述印记基因Z16为Snrpn/Snurf。
所述印记基因Z1(Gnas)，Z3(Peg10)，Z4(Igf2r)，Z5(Mest)，Z6(Plagl1)，Z8(Dcn)，Z10(Gatm)，Z11(Grb10)，Z13(Sgce)，Z16(Snrpn/Snurf)在正常肿瘤细胞组织内有不同程度的表达，在发生恶性病变时，表达量和印记状态都会发生 明显变化。
所述设计探针是根据印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16，即Gnas，Peg10，Igf2r，Mest，Plagl1，Dcn，Gatm，Grb10，Sgce和Snrpn/Snurf进行设计的，具体在每个基因的内旋子内选择一段序列作为探针，具体的探针由Advanced Cell Diagnostics公司设计。
在本文一实施例中，所述原位杂交采用RNAscope原位杂交方法。
在本文一实施例中，所述RNAscope原位杂交方法使用单通道或多通道的呈色试剂盒或者单通道或多通道的荧光试剂盒，优选为单通道红色/棕色呈色试剂盒或多通道的荧光试剂盒。
所述多通道呈色试剂盒或多通道荧光试剂盒包括两通道或两通道以上的呈色试剂盒或荧光试剂盒，所述两通道的呈色试剂盒或多通道的荧光试剂盒可以使用两个印记基因探针或印记基因和其他基因的联合表达甚至多个印记基因和非印记基因的综合表达。
在本文一实施例中，所述模型中的计算印记基因总表达量、印记基因缺失表达量和印记基因拷贝数异常表达量的公式如下：
总表达量＝(b+c+d)/(a+b+c+d)×100％；
正常印记基因总表达量＝b/(b+c+d)×100％；
印记基因缺失基因表达量(LOI)＝c/(b+c+d)×100％；
印记基因拷贝数异常的基因表达量(CNV)＝d/(b+c+d)×100％；
其中，所述a为将细胞进行苏木素染色后，细胞核内不存在标记，印记基因没有表达的细胞核；所述b为将细胞进行苏木素染色后，细胞核内存在一个红色/棕色标记，印记基因存在的细胞核；所述c为将细胞进行苏木素染色后，细胞核内存在两个红色/棕色标记，印记基因缺失的细胞核；所述d为将细胞进 行苏木素染色后，细胞核内存在两个以上红色/棕色标记，印记基因拷贝数异常的细胞核。
所述苏木素染色后的标记选自但不限于红色或棕色，用其他颜色进行染色标记也可用于印迹基因总表达量、印记基因缺失表达量和印记基因拷贝数异常表达量的计算。
将探针通过原位杂交，和Hemotoxy(苏木精)细胞核染色扩增信号，在40×或60×显微镜下，判断每一个细胞核内印记基因存在、印记基因缺失或拷贝数异常，通过计算印记基因总表达量、印记基因缺失基因表达量和印记基因拷贝数异常的基因表达量来判定该样本的肿瘤良恶性程度。由于切片仅为10微米，所以在显微镜下所见细胞核大约有20％为不完整细胞核，也就是说有部分假阴性的可能性存在。
在本文一实施例中，所述印记基因缺失表达量、印记基因拷贝数异常表达量和总表达量分成五个不同的等级。
所述五个不同的等级为在样本每个探针表达最阳性的区域对至少1200个细胞进行计数，针对Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16的十个印记基因的印记基因缺失表达量、印记基因拷贝数异常表达量和印记基因总表达量分别进行划分。
所述针对Z1的印记基因缺失表达量、印记基因拷贝数异常表达量和总表达量划分的五个不同的等级为：
0级：所述印记基因Z1的印记基因缺失表达量小于16％、所述印记基因Z1的印记基因拷贝数异常表达量小于2.5％或所述印记基因Z1的总表达量小于35％中的任意一种或至少两种的组合；
I级：所述印记基因Z1的印记基因缺失表达量为16-20％、所述印记基因 Z1的印记基因拷贝数异常表达量为2.5-4％或所述印记基因Z1的总表达量为35-40％中的任意一种或至少两种的组合；
II级：所述印记基因Z1的印记基因缺失表达量为20-25％、所述印记基因Z1的印记基因拷贝数异常表达量为4-6.5％或所述印记基因Z1的总表达量为40-50％中的任意一种或至少两种的组合；
III级：所述印记基因Z1的印记基因缺失表达量为25-30％、所述印记基因Z1的印记基因拷贝数异常表达量为6.5-8％或所述印记基因Z1的总表达量为50-65％中的任意一种或至少两种的组合；
IV级：所述印记基因Z1的印记基因缺失表达量大于30％、所述印记基因Z1的印记基因拷贝数异常表达量大于8％或所述印记基因Z1的总表达量大于65％中的任意一种或至少两种的组合。
所述针对Z3和Z13的印记基因缺失表达量、印记基因拷贝数异常表达量和总表达量划分的五个不同的等级为：
0级：所述印记基因Z3和Z13的印记基因缺失表达量小于10％、所述印记基因Z3和Z13的印记基因拷贝数异常表达量小于1％或所述印记基因Z3和Z13的总表达量小于25％中的任意一种或至少两种的组合；
I级：所述印记基因Z3和Z13的印记基因缺失表达量为10-15％、所述印记基因Z3和Z13的印记基因拷贝数异常表达量为1-2％或所述印记基因Z3和Z13的总表达量为25-30％中的任意一种或至少两种的组合；
II级：所述印记基因Z3和Z13的印记基因缺失表达量为15-25％、所述印记基因Z3和Z13的印记基因拷贝数异常表达量为2-3％或所述印记基因Z3和Z13的总表达量为30-45％中的任意一种或至少两种的组合；
III级：所述印记基因Z3和Z13的印记基因缺失表达量为25-30％、所述印 记基因Z3和Z13的印记基因拷贝数异常表达量为3-5％或所述印记基因Z3和Z13的总表达量为45-55％中的任意一种或至少两种的组合；
IV级：所述印记基因Z3和Z13的印记基因缺失表达量大于30％、所述印记基因Z3和Z13的印记基因拷贝数异常表达量大于5％或所述印记基因Z3和Z13的总表达量大于55％中的任意一种或至少两种的组合；
所述印记基因Z3和Z13的印记基因缺失表达量、印记基因拷贝数异常表达量和总表达量是相互独立的。
所述针对Z4、Z5、Z6、Z10和Z11的印记基因缺失表达量、印记基因拷贝数异常表达量和总表达量划分的五个不同的等级为：
0级：所述印记基因Z4、Z5、Z6、Z10和Z11的印记基因缺失表达量小于16％、所述印记基因Z4、Z5、Z6、Z10和Z11的印记基因拷贝数异常表达量小于1.5％或所述印记基因Z4、Z5、Z6、Z10和Z11的总表达量小于30％中的任意一种或至少两种的组合；
I级：所述印记基因Z4、Z5、Z6、Z10和Z11的印记基因缺失表达量为16-20％、所述印记基因Z4、Z5、Z6、Z10和Z11的印记基因拷贝数异常表达量为1.5-2.5％或所述印记基因Z4、Z5、Z6、Z10和Z11的总表达量为30-40％中的任意一种或至少两种的组合；
II级：所述印记基因Z4、Z5、Z6、Z10和Z11的印记基因缺失表达量为20-25％、所述印记基因Z4、Z5、Z6、Z10和Z11的印记基因拷贝数异常表达量为2.5-4％或所述印记基因Z4、Z5、Z6、Z10和Z11的总表达量为40-50％中的任意一种或至少两种的组合；
III级：所述印记基因Z4、Z5、Z6、Z10和Z11的印记基因缺失表达量为25-30％、所述印记基因Z4、Z5、Z6、Z10和Z11的印记基因拷贝数异常表达量 为4-6％或所述印记基因Z4、Z5、Z6、Z10和Z11的总表达量为50-65％中的任意一种或至少两种的组合；
IV级：所述印记基因Z4、Z5、Z6、Z10和Z11的印记基因缺失表达量大于30％、所述印记基因Z4、Z5、Z6、Z10和Z11的印记基因拷贝数异常表达量大于6％或所述印记基因Z4、Z5、Z6、Z10和Z11的总表达量大于65％中的任意一种或至少两种的组合；
所述印记基因Z4、Z5、Z6、Z10和Z11的印记基因缺失表达量、印记基因拷贝数异常表达量和总表达量是相互独立的。
用于检测食道肿瘤时，所述针对Z16的印记基因缺失表达量、印记基因拷贝数异常表达量和总表达量划分的五个不同的等级为：
0级：所述印记基因Z16的印记基因缺失表达量小于10％、所述印记基因Z16的印记基因拷贝数异常表达量小于1％或所述印记基因Z16的总表达量小于25％中的任意一种或至少两种的组合；
I级：所述印记基因Z16的印记基因缺失表达量为10-15％、所述印记基因Z16的印记基因拷贝数异常表达量为1-2％或所述印记基因Z16的总表达量为25-30％中的任意一种或至少两种的组合；
II级：所述印记基因Z16的印记基因缺失表达量为15-25％、所述印记基因Z16的印记基因拷贝数异常表达量为2-4％或所述印记基因Z16的总表达量为30-40％中的任意一种或至少两种的组合；
III级：所述印记基因Z16的印记基因缺失表达量为25-30％、所述印记基因Z16的印记基因拷贝数异常表达量为4-6％或所述印记基因Z16的总表达量为40-55％中的任意一种或至少两种的组合；
IV级：所述印记基因Z16的印记基因缺失表达量大于30％、所述印记基因 Z16的印记基因拷贝数异常表达量大于6％或所述印记基因Z16的总表达量大于55％中的任意一种或至少两种的组合。
用于检测胃肿瘤时，所述针对Z16的印记基因缺失表达量、印记基因拷贝数异常表达量和总表达量划分的五个不同的等级为：
0级：所述印记基因Z16的印记基因缺失表达量小于15％、所述印记基因Z16的印记基因拷贝数异常表达量小于1.5％或所述印记基因Z16的总表达量小于30％中的任意一种或至少两种的组合；
I级：所述印记基因Z16的印记基因缺失表达量为15-20％、所述印记基因Z16的印记基因拷贝数异常表达量为1.5-4％或所述印记基因Z16的总表达量为30-40％中的任意一种或至少两种的组合；
II级：所述印记基因Z16的印记基因缺失表达量为20-25％、所述印记基因Z16的印记基因拷贝数异常表达量为4-7％或所述印记基因Z16的总表达量为40-50％中的任意一种或至少两种的组合；
III级：所述印记基因Z16的印记基因缺失表达量为25-30％、所述印记基因Z16的印记基因拷贝数异常表达量为7-10％或所述印记基因Z16的总表达量为50-60％中的任意一种或至少两种的组合；
IV级：所述印记基因Z16的印记基因缺失表达量大于30％、所述印记基因Z16的印记基因拷贝数异常表达量大于10％或所述印记基因Z16的总表达量大于60％中的任意一种或至少两种的组合。
由于印记基因Z16在胃癌中表达的特殊性，Z16的III级和IV级大部分出现在胃癌的早中期，90％的晚期胃癌样本Z16表达都较低。
用于检测食道肿瘤时，所述针对Z8的印记基因缺失表达量、印记基因拷贝数异常表达量和总表达量划分的五个不同的等级为：
0级：所述印记基因Z8的印记基因缺失表达量小于16％、所述印记基因Z8的印记基因拷贝数异常表达量小于2.5％或所述印记基因Z8的总表达量小于35％中的任意一种或至少两种的组合；
I级：所述印记基因Z8的印记基因缺失表达量为16-20％、所述印记基因Z8的印记基因拷贝数异常表达量为2.5-4％或所述印记基因Z8的总表达量为35-40％中的任意一种或至少两种的组合；
II级：所述印记基因Z8的印记基因缺失表达量为20-25％、所述印记基因Z8的印记基因拷贝数异常表达量为4-6.5％或所述印记基因Z8的总表达量为40-50％中的任意一种或至少两种的组合；
III级：所述印记基因Z8的印记基因缺失表达量为25-30％、所述印记基因Z8的印记基因拷贝数异常表达量为6.5-8％或所述印记基因Z8的总表达量为50-65％中的任意一种或至少两种的组合；
IV级：所述印记基因Z8的印记基因缺失表达量大于30％、所述印记基因Z8的印记基因拷贝数异常表达量大于8％或所述印记基因Z8的总表达量大于65％中的任意一种或至少两种的组合；
用于检测胃肿瘤时，所述针对Z8的印记基因缺失表达量、印记基因拷贝数异常表达量和总表达量划分的五个不同的等级为：
0级：所述印记基因Z8的印记基因缺失表达量小于10％、所述印记基因Z8的印记基因拷贝数异常表达量小于1％或所述印记基因Z8的总表达量小于25％中的任意一种或至少两种的组合；
I级：所述印记基因Z8的印记基因缺失表达量为10-15％、所述印记基因Z8的印记基因拷贝数异常表达量为1-2％或所述印记基因Z8的总表达量为25-30％中的任意一种或至少两种的组合；
II级：所述印记基因Z8的印记基因缺失表达量为15-25％、所述印记基因Z8的印记基因拷贝数异常表达量为2-3％或所述印记基因Z8的总表达量为30-45％中的任意一种或至少两种的组合；
III级：所述印记基因Z8的印记基因缺失表达量为25-30％、所述印记基因Z8的印记基因拷贝数异常表达量为3-5％或所述印记基因Z8的总表达量为45-55％中的任意一种或至少两种的组合；
IV级：所述印记基因Z8的印记基因缺失表达量大于30％、所述印记基因Z8的印记基因拷贝数异常表达量大于5％或所述印记基因Z8的总表达量大于55％中的任意一种或至少两种的组合。
发明人发现印记基因Z8和Z16比较特殊，这两个基因在食道肿瘤或胃肿瘤中的印记基因缺失表达量、印记基因拷贝数异常表达量和总表达量的分级情况是不同的，所以将印记基因Z8和Z16按照食道肿瘤或胃肿瘤两种情况分开来分级。
印记基因Z16在胃肿瘤组织间质和上皮细胞中均匀表达，但在临床分期II期以前的早期胃癌和临床分期III-IV期的中晚期胃癌中表达有明显的不同。
在本文一实施例中，诊断食道肿瘤的良恶性程度分为良性肿瘤、食道癌潜能、早期食道癌、中期食道癌和晚期食道癌。
在本文一实施例中，所述诊断食道的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16的印记基因缺失表达量和印记基因拷贝数异常表达量均小于I级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因缺失表达量为I级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因拷贝数异常表达量为I级中的任意一种情况，则为良性肿瘤；
所述诊断食道的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的至少2个印记基因的印记基因缺失表达量为I级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16的至少2个印记基因的印记基因拷贝数异常表达量为I级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中不超过1个印记基因的印记基因缺失表达量为II级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因拷贝数异常表达量为II级中的任意一种情况，则为食道癌潜能；
所述诊断食道的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的至少2个印记基因的印记基因缺失表达量为II级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16的至少2个印记基因的印记基因拷贝数异常表达量为II级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中不超过1个印记基因的印记基因缺失表达量为III级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因拷贝数异常表达量为III级中的任意一种情况，则为早期食道癌；
所述诊断食道的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的至少2个印记基因的印记基因缺失表达量为III级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16的至少2个印记基因的印记基因拷贝数异常表达量为III级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中不超过1个印记基因的印记基因缺失表达量为IV级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因拷贝数异常表达量为IV级中的任意一种情况，则 为中期食道癌；
所述诊断食道的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中至少2个印记基因的印记基因缺失表达量为IV级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中至少2个印记基因的印记基因拷贝数异常表达量为IV级，则为晚期食道癌。
在本文一实施例中，诊断胃肿瘤的良恶性程度分为良性胃肿瘤、胃癌潜能、早期胃癌、中期胃癌和晚期胃癌。
所述诊断胃肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16的印记基因缺失表达量和印记基因拷贝数异常表达量均小于I级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因缺失表达量为I级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因拷贝数异常表达量为I级中的任意一种情况，则为良性肿瘤；
所述诊断胃肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的至少2个印记基因的印记基因缺失表达量为I级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13的至少2个印记基因的印记基因拷贝数异常表达量为I级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的不超过1个印记基因的印记基因缺失表达量为I级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的不超过1个印记基因的印记基因拷贝数异常表达量为I级且印记基因Z16的印记基因缺失表达量或印记基因拷贝数异常表达量为I级或以上，或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中不超过1个印记基因的印记基因缺失表达量为II级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的不超过1个印记基因的印 记基因拷贝数异常表达量为II级中的任意一种情况，则为胃癌潜能；
所述诊断胃肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的至少2个印记基因的印记基因缺失表达量为II级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的至少2个印记基因的印记基因拷贝数异常表达量为II级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中不超过1个印记基因的印记基因缺失表达量为III级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的不超过1个印记基因的印记基因拷贝数异常表达量为III级中的任意一种情况，则为早期胃癌；
所述诊断胃肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的至少2个印记基因的印记基因缺失表达量为III级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13的至少2个印记基因的印记基因拷贝数异常表达量为III级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中不超过1个印记基因的印记基因缺失表达量为IV级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的不超过1个印记基因的印记基因拷贝数异常表达量为IV级中的任意一种情况，则为中期胃癌；
所述诊断胃肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中至少2个印记基因的印记基因缺失表达量为IV级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中至少2个印记基因的印记基因拷贝数异常表达量为IV级，则为晚期胃癌。
在本文一实施例中，本公开提供一种所述的印记基因分级模型和/或所述的装置用于食道肿瘤和胃肿瘤检测。
在本文一实施例中，本公开提供一种所述的印记基因分级模型和/或所述的装置用于制备治疗食道肿瘤和/或胃肿瘤的药物或器械的用途。
在本文一实施例中，诊断食道肿瘤的良恶性程度分为良性肿瘤、食道癌潜能、早期食道癌、中期食道癌和晚期食道癌。
所述诊断食道肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16的印记基因缺失表达量和印记基因拷贝数异常表达量均小于I级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因缺失表达量为I级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因拷贝数异常表达量为I级中的任意一种情况，则为良性肿瘤；
所述诊断食道肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的至少2个印记基因的印记基因缺失表达量为I级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16的至少2个印记基因的印记基因拷贝数异常表达量为I级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中不超过1个印记基因的印记基因缺失表达量为II级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因拷贝数异常表达量为II级中的任意一种情况，则为食道癌潜能；
所述诊断食道肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的至少2个印记基因的印记基因缺失表达量为II级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16的至少2个印记基因的印记基因拷贝数异常表达量为II级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中不超过1个印记基因的印记基因缺失表达量为III级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因拷贝数异常表达量为III级中的任意一种情 况，则为早期食道癌；
所述诊断食道肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的至少2个印记基因的印记基因缺失表达量为III级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16的至少2个印记基因的印记基因拷贝数异常表达量为III级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中不超过1个印记基因的印记基因缺失表达量为IV级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因拷贝数异常表达量为IV级中的任意一种情况，则为中期食道癌；
所述诊断食道肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中至少2个印记基因的印记基因缺失表达量为IV级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中至少2个印记基因的印记基因拷贝数异常表达量为IV级，则为晚期食道癌。
在本文一实施例中，所述诊断胃肿瘤的良恶性程度分为良性胃肿瘤、胃癌潜能、早期胃癌、中期胃癌和晚期胃癌。
所述诊断胃肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16的印记基因缺失表达量和印记基因拷贝数异常表达量均小于I级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因缺失表达量为I级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因拷贝数异常表达量为I级中的任意一种情况，则为良性肿瘤；
所述诊断胃肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的至少2个印记基因的印记基因缺失表达量为I级，印 记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13的至少2个印记基因的印记基因拷贝数异常表达量为I级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的不超过1个印记基因的印记基因缺失表达量为I级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的不超过1个印记基因的印记基因拷贝数异常表达量为I级且印记基因Z16的印记基因缺失表达量或印记基因拷贝数异常表达量为I级或以上，或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中不超过1个印记基因的印记基因缺失表达量为II级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的不超过1个印记基因的印记基因拷贝数异常表达量为II级中的任意一种情况，则为胃癌潜能；
所述诊断胃肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的至少2个印记基因的印记基因缺失表达量为II级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的至少2个印记基因的印记基因拷贝数异常表达量为II级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中不超过1个印记基因的印记基因缺失表达量为III级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的不超过1个印记基因的印记基因拷贝数异常表达量为III级中的任意一种情况，则为早期胃癌；
所述诊断胃肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的至少2个印记基因的印记基因缺失表达量为III级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13的至少2个印记基因的印记基因拷贝数异常表达量为III级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中不超过1个印记基因的印记基因缺失表达量为IV级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的不超过1个印记基因的印记基因拷贝数异常表达量为IV级中的任意一种情况，则为中期胃癌；
所述诊断胃肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中至少2个印记基因的印记基因缺失表达量为IV级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中至少2个印记基因的印记基因拷贝数异常表达量为IV级，则为晚期胃癌。
在一个具体的实施例中，本公开提供一种晚期胃癌诊断的标志物，所述标志物为印记基因Z1和印记基因Z11；
其中，所述印记基因Z1为Gnas，所述印记基因Z11为Grb10；
在一个具体的实施例中，所述晚期胃癌为胃癌的TNM等级大于T2N0M0的胃癌。
印记基因Z16在胃肿瘤组织间质和上皮细胞中均匀表达，但在临床分期II期以前的早期胃癌和临床分期III-IV期的中晚期胃癌中表达有明显的不同。在早期胃癌中，特别是在上皮内瘤变时期，75％的样本印记基因Z16的印记缺失、拷贝数异常和总表达量达到I级以上，随着癌症的恶化，90％的中晚期胃癌样本印记基因Z16的印记缺失、拷贝数异常和总表达量为0级。所以印记基因Z16联合印记基因Z1和Z11，对早期胃癌的诊断，特别是低级别上皮内瘤变到胃癌的转变过程，是一个很好的诊断标记物，但在晚期胃癌中印记基因Z16则用处不大，仅需要Z1和Z11就能作出诊断。
与相关技术相比，本文实施例中，利用所述检测模型和装置，以直观的方法表现了印记基因在食道肿瘤和/或胃肿瘤病人样本上的表现，通过对印记基因原位标记的方法，客观，直观，早期，精确地检测出印记(迹)基因的变化，并可以提供量化的模型，为分子病理学的诊断做出巨大贡献。
附图说明
图1是本公开苏木素染色细胞核的胃癌的病理切片，其中，所述a为将细 胞进行苏木素染色后，细胞核内不存在标记，印记基因没有表达；所述b为将细胞进行苏木素染色后，细胞核内存在一个红色/棕色标记，印记基因存在；所述c为将细胞进行苏木素染色后，细胞核内存在两个红色/棕色标记，印记基因缺失；所述d为将细胞进行苏木素染色后，细胞核内存在两个以上红色/棕色标记，印记基因拷贝数异常；
图2(a)为0级胃肿瘤的病理切片中10个基因的表达状态，图2(b)为I级胃癌的病理切片中10个基因的表达状态，图2(c)为II级胃癌的病理切片中10个基因的表达状态，图2(d)为III级胃癌的病理切片中10个基因的表达状态，图2(e)为IV级胃癌的病理切片中10个基因的表达状态；
图3(a)为印记基因Z1、Z11和Z16对胃癌的印记缺失的强度，图3(b)为印记基因Z1、Z11和Z16对胃癌的拷贝数异常的强度，图3(c)为印记基因Z1、Z11和Z16对胃癌的总表达量的强度，图3(d)为印记基因Z3、Z4、Z5、Z6、Z8、Z10和Z13对胃癌的印记缺失的强度，图3(e)为印记基因Z3、Z4、Z5、Z6、Z8、Z10和Z13对胃癌的拷贝数异常的强度，图3(f)为印记基因Z3、Z4、Z5、Z6、Z8、Z10和Z13对胃癌的总表达量的强度，其中，LOI为印记基因缺失基因表达量，CNV为印记基因拷贝数异常的基因表达量，TE为印记基因总表达量；
图4(a)为印记基因Z1印记缺失、拷贝数异常和总表达量的强度，图4(b)为印记基因Z11印记缺失、拷贝数异常和总表达量的强度，图4(c)为印记基因Z16印记缺失、拷贝数异常和总表达量的强度，图4(d)为印记基因Z3印记缺失、拷贝数异常和总表达量的强度，图4(e)为印记基因Z4印记缺失、拷贝数异常和总表达量的强度，图4(f)为印记基因Z5印记缺失、拷贝数异常和总表达量的强度，图4(g)为印记基因Z6印记缺失、拷贝数异常和总表达量的 强度，图4(h)为印记基因Z8印记缺失、拷贝数异常和总表达量的强度，图4(i)为印记基因Z10印记缺失、拷贝数异常和总表达量的强度，图4(j)为印记基因Z13印记缺失、拷贝数异常和总表达量的强度，其中，LOI为印记基因缺失基因表达量，CNV为印记基因拷贝数异常的基因表达量，TE为印记基因总表达量；
图5(a)为印记基因Z1应用于37例胃肿瘤病理切片中，印记缺失和拷贝数异常的分布范围和分级标准，图5(b)为印记基因Z11应用于37例胃肿瘤病理切片中，印记缺失和拷贝数异常的分布范围和分级标准，图5(c)为印记基因Z16应用于37例胃肿瘤病理切片中，印记缺失和拷贝数异常的分布范围和分级标准，图5(d)为印记基因Z3应用于37例胃肿瘤病理切片中，印记缺失和拷贝数异常的分布范围和分级标准，图5(e)为印记基因Z4应用于37例胃肿瘤病理切片中，印记缺失和拷贝数异常的分布范围和分级标准，图5(f)为印记基因Z5应用于37例胃肿瘤病理切片中，印记缺失和拷贝数异常的分布范围和分级标准，图5(g)为印记基因Z6应用于37例胃肿瘤病理切片中，印记缺失和拷贝数异常的分布范围和分级标准，图5(h)为印记基因Z8应用于37例胃肿瘤病理切片中，印记缺失和拷贝数异常的分布范围和分级标准，图5(i)为印记基因Z10应用于37例胃肿瘤病理切片中，印记缺失和拷贝数异常的分布范围和分级标准，图5(j)为印记基因Z13应用于37例胃肿瘤病理切片中，印记缺失和拷贝数异常的分布范围和分级标准，其中，LOI为印记基因缺失基因表达量，CNV为印记基因拷贝数异常的基因表达量，TE为印记基因总表达量；
图6(a)为0级食道肿瘤的病理切片中10个基因的表达状态，图6(b)为I级食道癌的病理切片中10个基因的表达状态，图6(c)为II级食道癌的病理切片中10个基因的表达状态，图6(d)为III级食道癌的病理切片中10个基 因的表达状态，图6(e)为IV级食道癌的病理切片中10个基因的表达状态；
图7(a)为印记基因Z1、Z11和Z16对食道癌的印记缺失的强度，图7(b)为印记基因Z1、Z11和Z16对食道癌的拷贝数异常的强度，图7(c)为印记基因Z1、Z11和Z16对食道癌的总表达量的强度，图7(d)为印记基因Z3、Z4、Z5、Z6、Z8、Z10和Z13对食道癌的印记缺失的强度，图7(e)为印记基因Z3、Z4、Z5、Z6、Z8、Z10和Z13对食道癌的拷贝数异常的强度，图7(f)为印记基因Z3、Z4、Z5、Z6、Z8、Z10和Z13对食道癌的总表达量的强度，其中，LOI为印记基因缺失基因表达量，CNV为印记基因拷贝数异常的基因表达量，TE为印记基因总表达量；
其中图8(a)为印记基因Z1印记缺失、拷贝数异常和总表达量的强度，图8(b)为印记基因Z11印记缺失、拷贝数异常和总表达量的强度，图8(c)为印记基因Z16印记缺失、拷贝数异常和总表达量的强度，图8(d)为印记基因Z3印记缺失、拷贝数异常和总表达量的强度，图8(e)为印记基因Z4印记缺失、拷贝数异常和总表达量的强度，图8(f)为印记基因Z5印记缺失、拷贝数异常和总表达量的强度，图8(g)为印记基因Z6印记缺失、拷贝数异常和总表达量的强度，图8(h)为印记基因Z8印记缺失、拷贝数异常和总表达量的强度，图8(i)为印记基因Z10印记缺失、拷贝数异常和总表达量的强度，图8(j)为印记基因Z13印记缺失、拷贝数异常和总表达量的强度，其中，LOI为印记基因缺失基因表达量，CNV为印记基因拷贝数异常的基因表达量，TE为印记基因总表达量；
图9(a)为印记基因Z1应用于57例食道肿瘤病理切片中，印记缺失和拷贝数异常的分布范围和分级标准，图9(b)为印记基因Z11应用于57例食道肿瘤病理切片中，印记缺失和拷贝数异常的分布范围和分级标准，图9(c)为印 记基因Z16应用于57例食道肿瘤病理切片中，印记缺失和拷贝数异常的分布范围和分级标准，图9(d)为印记基因Z3应用于57例食道肿瘤病理切片中，印记缺失和拷贝数异常的分布范围和分级标准，图9(e)为印记基因Z4应用于57例食道肿瘤病理切片中，印记缺失和拷贝数异常的分布范围和分级标准，图9(f)为印记基因Z5应用于57例食道肿瘤病理切片中，印记缺失和拷贝数异常的分布范围和分级标准，图9(g)为印记基因Z6应用于57例食道肿瘤病理切片中，印记缺失和拷贝数异常的分布范围和分级标准，图9(h)为印记基因Z8应用于57例食道肿瘤病理切片中，印记缺失和拷贝数异常的分布范围和分级标准，图9(i)为印记基因Z10应用于57例食道肿瘤病理切片中，印记缺失和拷贝数异常的分布范围和分级标准，图9(j)为印记基因Z13应用于57例食道肿瘤病理切片中，印记缺失和拷贝数异常的分布范围和分级标准，其中，LOI为印记基因缺失基因表达量，CNV为印记基因拷贝数异常的基因表达量，TE为印记基因总表达量。
具体实施方式
为更进一步阐述本公开所采取的技术手段及其效果，以下结合附图并通过具体实施方式来进一步说明本公开的技术方案，但本公开并非局限在实施例范围内。
基因组印记是表观遗传学中基因调控的一种方式。其特点是，通过甲基化来自特定亲代的等位基因，使某个基因只有一个等位基因表达，而另一个则陷入基因沉默状态。该种类的基因，被称为印迹(记)基因。印迹缺失是印迹基因去甲基化导致沉默状态的等位基因被激活并且开始基因表达的一种表观遗传改变。大量研究表明，该现象(印迹缺失)普遍存在于各类癌症并且发生时间早于细胞和组织形态改变。与此同时，在健康细胞中，印迹缺失比例极低，与 癌细胞成鲜明对比。所以，印迹基因的甲基化状态可以作为病理标记，通过特定分子检测技术，对细胞异常状态进行分析。
本公开所述检测模型和系统，以直观的方法表现了印记缺失在食道和胃肿瘤病人的样本上的表现，通过对印记基因原位标记的方法，客观，直观，早期，精确地检测出印记(迹)基因的变化，并可以提供量化的模型，为食道和胃肿瘤的诊断做出巨大贡献；
本公开检测装置，可以在食道和胃肿瘤病人手术前通过胃镜活检样本得出食道和胃肿瘤良恶性程度以及癌细胞侵袭范围的判断，从而为手术及精准治疗提供依据，这是细胞分子领域诊断食道和胃肿瘤的革命性突破；
本公开可以精确的判断食道和胃肿瘤的级别，通过印记基因的组合检测对食道和胃肿瘤的早期病变即上皮内瘤变的恶性程度进行明确地分级，特别是能够对早期和中晚期胃癌进行准确区分，极大地提高了对食道癌和胃癌的早期、明确诊断，特别是用在早期普查和癌症术后随访，尤其是对于疑似复发病人的跟踪随访，可以争取时间，为挽救病人生命做出重大贡献；
本公开检测方法区别于免疫组化方法，减少了假阳性和其他负面作用，不仅如此，通过发现的食道和胃肿瘤相关印记基因缺失位点的致该基因沉默、剔除、重排的靶向药物或技术方法，可用于指导后期的治疗和用药。
实施例1 胃肿瘤的印记基因分析
所述的印记基因的检测方法，包括如下步骤：
(1)获取胃肿瘤的组织细胞切片(10微米)，放入10％中性福尔马林溶液中进行固定，以防RNA降解，固定时间为24小时，石蜡包埋(FFPE)，所述玻片需要用正电荷脱载玻片，所述切片在40℃烤箱烘烤3h以上；
(2)按照RNASCope的样品处理方法进行脱蜡处理，封闭样本中内源性过 氧化物酶活性，增强通透性并暴露出RNA分子；
(3)设计探针：根据印记基因序列设计特异性引物；
所述设计探针是根据印记基因Z1(Gnas)、Z3(Peg10)、Z4(Igf2r)、Z5(Mest)、Z6(Plagl1)、Z8(Dcn)、Z10(Gatm)、Z11(Grb10)、Z13(Sgce)和Z16(Snrpn/Snurf)进行设计的，具体在每个基因的内旋子内选择一段序列作为探针，具体的探针由Advanced Cell Diagnostics公司设计。
(4)将步骤(3)的探针与待测样本通过试剂盒进行RNA SCope原位杂交；
(5)信号扩增和苏木精染色，用显微镜成像分析印记基因的表达情况；
所述模型中的计算印记基因总表达量、印记基因缺失表达量和印记基因拷贝数异常表达量的公式如下：
总表达量＝(b+c+d)/(a+b+c+d)×100％；
正常印记基因总表达量＝b/(b+c+d)×100％；
印记基因缺失基因表达量(LOI)＝c/(b+c+d)×100％；
印记基因拷贝数异常的基因表达量(CNV)＝d/(b+c+d)×100％；
其中，a、b、c、d如图1所示，所述a为将细胞进行苏木素染色后，细胞核内不存在标记，印记基因没有表达的细胞核；所述b为将细胞进行苏木素染色后，细胞核内存在一个红色/棕色标记，印记基因存在的细胞核；所述c为将细胞进行苏木素染色后，细胞核内存在两个红色/棕色标记，印记基因缺失的细胞核；所述d为将细胞进行苏木素染色后，细胞核内存在两个以上红色/棕色标记，印记基因拷贝数异常的细胞核。
从图2(a)-图2(e)可以看出，从0级到IV级的样本中，印记缺失(细胞核内有两个信号点)和拷贝数异常(细胞核内有三个或以上信号点)的细胞比例随恶性程度的增加而逐渐增加。
实施例2 胃镜活检样本的印记基因分析
所述胃镜活检样本是，在胃镜下取出可疑病变组织，10％中性福尔马林溶液固定24h以上，其他检测方法同实施例1。
从图3(a)-图3(f)可以看出，Z1，Z3，Z4，Z5，Z6，Z8，Z10，Z11，Z13，Z16每个基因对胃癌的反应敏感性或者说对应于胃癌表达的印记缺失的强度和状态是不同的。
具体每个印记基因对胃癌的敏感度如图4(a)-图4(j)，从图4(a)-图4(c)可以看出，印记基因Z1的印记缺失和表达量增加在恶性潜能阶段开始出现，在早期到晚期的胃癌发展过程中快速上升到很高水平，印记基因Z1的拷贝数异常在恶性潜能阶段开始出现，在早期和中期胃癌中快速上升，到晚期胃癌阶段上升速度减缓；印记基因Z11的印记缺失在恶性潜能阶段开始出现，在早期胃癌阶段快速上升，在中期胃癌中上升速度减缓，到晚期胃癌阶段又继续上升到较高水平，印记基因Z11的拷贝数异常和表达量增加在恶性潜能阶段开始出现，在早期到晚期的胃癌发展过程中逐渐上升到较高水平；印记基因Z16的印记缺失、拷贝数异常和表达量增加在恶性潜能和早期胃癌阶段快速上升，但是到中晚期胃癌阶段却出现明显下降；
从图4(d)-图4(j)可以看出，印记基因Z3的印记缺失、拷贝数异常和表达量增加在早期胃癌阶段开始出现，在中期胃癌中继续上升，在晚期胃癌中上升速度减缓；印记基因Z4的印记缺失在早期胃癌阶段开始出现，在中期胃癌阶段快速上升，到晚期胃癌阶段上升速度减缓，印记基因Z4的拷贝数异常在早期胃癌阶段快速上升，在中期和晚期胃癌阶段上升速度减缓，印记基因Z4的表达量增加在恶性潜能阶段开始出现，在早期胃癌阶段快速上升，中期胃癌阶段没有明显上升，到晚期胃癌阶段又继续上升；印记基因Z5的印记缺失在早期胃 癌阶段快速上升，在中期胃癌阶段上升速度减缓，到晚期胃癌阶段又继续上升到较高水平，印记基因Z5的拷贝数异常在早期胃癌阶段开始出现，在中期胃癌中快速上升，在晚期胃癌阶段上升速度减缓，印记基因Z5的表达量增加在早期胃癌阶段开始出现，在中期和晚期胃癌中逐渐上升到较高水平；印记基因Z6的印记缺失在早期胃癌阶段开始出现，在中期胃癌中继续上升，到晚期胃癌阶段上升速度减缓，印记基因Z6的拷贝数异常在早期胃癌阶段开始出现，在中期胃癌中上升速度减缓，到晚期胃癌阶段又继续上升，印记基因Z6的表达量增加在早期胃癌阶段开始出现，在中期和晚期胃癌中缓慢上升；印记基因Z8的印记缺失和表达量增加在早期胃癌中开始出现，在中期胃癌中缓慢上升，在晚期胃癌阶段上升速度增加，但水平仍然不高，印记基因Z8的拷贝数异常在早期胃癌阶段快速上升，但是在中期和晚期胃癌中没有继续增加；印记基因Z10的印记缺失、拷贝数异常和表达量增加在恶性潜能阶段开始出现，在早期到晚期的胃癌发展过程中逐渐上升到较高水平；印记基因Z13的印记缺失和表达量增加在恶性潜能阶段开始出现，在早期和中期胃癌中逐渐上升，在晚期胃癌中上升速度减缓，印记基因Z13的拷贝数异常在恶性潜能阶段开始出现，在早期胃癌中上升速度减缓，中期胃癌阶段上升速度增加，在晚期胃癌中上升速度又减缓。
实施例3 37例胃肿瘤样本的印记基因分析
获取37例胃肿瘤病人的组织包括胃镜活检样本(10微米)，检测方法同实施例1。
从图5(a)-图5(j)可以看出，37例胃肿瘤组织样本中Z1，Z3，Z4，Z5，Z6，Z8，Z10，Z11，Z13和Z16十个探针的印记缺失和拷贝数异常的比例呈现从低到高的分布，根据不同探针的分布趋势，我们计算得到了图中虚线所示的分级标准，可以将每个探针的印记缺失和拷贝数异常分别从低到高分成5个等 级。
具体的分级如下：
从图5(a)可以看出，对于所述印记基因Z1，印记基因缺失表达量小于16％、印记基因拷贝数异常表达量小于2.5％或印记基因总表达量小于35％中的任意一种或至少两种的组合为0级，印记基因缺失表达量为16-20％、印记基因拷贝数异常表达量为2.5-4％或印记基因总表达量为35-40％中的任意一种或至少两种的组合为I级，印记基因缺失表达量为20-25％、印记基因拷贝数异常表达量为4-6.5％或印记基因总表达量为40-50％中的任意一种或至少两种的组合为II级，印记基因缺失表达量为25-30％、印记基因拷贝数异常表达量为6.5-8％或印记基因总表达量为50-65％中的任意一种或至少两种的组合为III级，印记基因缺失表达量大于30％、印记基因拷贝数异常表达量大于8％或印记基因总表达量大于65％中的任意一种或至少两种的组合为IV级；
从图5(b)可以看出，对于所述印记基因Z11，印记基因缺失表达量小于16％、印记基因拷贝数异常表达量小于1.5％或印记基因总表达量小于30％中的任意一种或至少两种的组合为0级，印记基因缺失表达量为16-20％、印记基因拷贝数异常表达量为1.5-2.5％或印记基因总表达量为30-40％中的任意一种或至少两种的组合为I级，印记基因缺失表达量为20-25％、印记基因拷贝数异常表达量为2.5-4％或印记基因总表达量为40-50％中的任意一种或至少两种的组合为II级，印记基因缺失表达量为25-30％、印记基因拷贝数异常表达量为4-6％或印记基因总表达量为50-65％中的任意一种或至少两种的组合为III级，印记基因缺失表达量大于30％、印记基因拷贝数异常表达量大于6％或印记基因总表达量大于65％中的任意一种或至少两种的组合为IV级；
从图5(c)可以看出，对于所述印记基因Z16，印记基因缺失表达量小于 15％、印记基因拷贝数异常表达量小于1.5％或印记基因总表达量小于30％中的任意一种或至少两种的组合为0级，印记基因缺失表达量为15-20％、印记基因拷贝数异常表达量为1.5-4％或印记基因总表达量为30-40％中的任意一种或至少两种的组合为I级，印记基因缺失表达量为20-25％、印记基因拷贝数异常表达量为4-7％或印记基因总表达量为40-50％中的任意一种或至少两种的组合为II级，印记基因缺失表达量为25-30％、印记基因拷贝数异常表达量为7-10％或印记基因总表达量为50-60％中的任意一种或至少两种的组合为III级，印记基因缺失表达量大于30％、印记基因拷贝数异常表达量大于10％或印记基因总表达量大于60％中的任意一种或至少两种的组合为IV级；
从图5(d)可以看出，对于所述印记基因Z3，印记基因缺失表达量小于10％、印记基因拷贝数异常表达量小于1％或印记基因总表达量小于25％中的任意一种或至少两种的组合为0级，印记基因缺失表达量为10-15％、印记基因拷贝数异常表达量为1-2％或印记基因总表达量为25-30％中的任意一种或至少两种的组合为I级，印记基因缺失表达量为15-25％、印记基因拷贝数异常表达量为2-3％或印记基因总表达量为30-45％中的任意一种或至少两种的组合为II级，印记基因缺失表达量为25-30％、印记基因拷贝数异常表达量为3-5％或印记基因总表达量为45-55％中的任意一种或至少两种的组合为III级，印记基因缺失表达量大于30％、印记基因拷贝数异常表达量大于5％或印记基因总表达量大于55％中的任意一种或至少两种的组合为IV级；
从图5(e)可以看出，对于所述印记基因Z4，印记基因缺失表达量小于16％、印记基因拷贝数异常表达量小于1.5％或印记基因总表达量小于30％中的任意一种或至少两种的组合为0级，印记基因缺失表达量为16-20％、印记基因拷贝数异常表达量为1.5-2.5％或印记基因总表达量为30-40％中的任意一种或至少两种 的组合为I级，印记基因缺失表达量为20-25％、印记基因拷贝数异常表达量为2.5-4％或印记基因总表达量为40-50％中的任意一种或至少两种的组合为II级，印记基因缺失表达量为25-30％、印记基因拷贝数异常表达量为4-6％或印记基因总表达量为50-65％中的任意一种或至少两种的组合为III级，印记基因缺失表达量大于30％、印记基因拷贝数异常表达量大于6％或印记基因总表达量大于65％中的任意一种或至少两种的组合为IV级；
从图5(f)可以看出，对于所述印记基因Z5，印记基因缺失表达量小于16％、印记基因拷贝数异常表达量小于1.5％或印记基因总表达量小于30％中的任意一种或至少两种的组合为0级，印记基因缺失表达量为16-20％、印记基因拷贝数异常表达量为1.5-2.5％或印记基因总表达量为30-40％中的任意一种或至少两种的组合为I级，印记基因缺失表达量为20-25％、印记基因拷贝数异常表达量为2.5-4％或印记基因总表达量为40-50％中的任意一种或至少两种的组合为II级，印记基因缺失表达量为25-30％、印记基因拷贝数异常表达量为4-6％或印记基因总表达量为50-65％中的任意一种或至少两种的组合为III级，印记基因缺失表达量大于30％、印记基因拷贝数异常表达量大于6％或印记基因总表达量大于65％中的任意一种或至少两种的组合为IV级；
从图5(g)可以看出，对于所述印记基因Z6，印记基因缺失表达量小于16％、印记基因拷贝数异常表达量小于1.5％或印记基因总表达量小于30％中的任意一种或至少两种的组合为0级，印记基因缺失表达量为16-20％、印记基因拷贝数异常表达量为1.5-2.5％或印记基因总表达量为30-40％中的任意一种或至少两种的组合为I级，印记基因缺失表达量为20-25％、印记基因拷贝数异常表达量为2.5-4％或印记基因总表达量为40-50％中的任意一种或至少两种的组合为II级，印记基因缺失表达量为25-30％、印记基因拷贝数异常表达量为4-6％或印记基因 总表达量为50-65％中的任意一种或至少两种的组合为III级，印记基因缺失表达量大于30％、印记基因拷贝数异常表达量大于6％或印记基因总表达量大于65％中的任意一种或至少两种的组合为IV级；
从图5(h)可以看出，对于所述印记基因Z8，印记基因缺失表达量小于10％、印记基因拷贝数异常表达量小于1％或印记基因总表达量小于25％中的任意一种或至少两种的组合为0级，印记基因缺失表达量为10-15％、印记基因拷贝数异常表达量为1-2％或印记基因总表达量为25-30％中的任意一种或至少两种的组合为I级，印记基因缺失表达量为15-25％、印记基因拷贝数异常表达量为2-3％或印记基因总表达量为30-45％中的任意一种或至少两种的组合为II级，印记基因缺失表达量为25-30％、印记基因拷贝数异常表达量为3-5％或印记基因总表达量为45-55％中的任意一种或至少两种的组合为III级，印记基因缺失表达量大于30％、印记基因拷贝数异常表达量大于5％或印记基因总表达量大于55％中的任意一种或至少两种的组合为IV级；
从图5(i)可以看出，对于所述印记基因Z10，印记基因缺失表达量小于16％、印记基因拷贝数异常表达量小于1.5％或印记基因总表达量小于30％中的任意一种或至少两种的组合为0级，印记基因缺失表达量为16-20％、印记基因拷贝数异常表达量为1.5-2.5％或印记基因总表达量为30-40％中的任意一种或至少两种的组合为I级，印记基因缺失表达量为20-25％、印记基因拷贝数异常表达量为2.5-4％或印记基因总表达量为40-50％中的任意一种或至少两种的组合为II级，印记基因缺失表达量为25-30％、印记基因拷贝数异常表达量为4-6％或印记基因总表达量为50-65％中的任意一种或至少两种的组合为III级，印记基因缺失表达量大于30％、印记基因拷贝数异常表达量大于6％或印记基因总表达量大于65％中的任意一种或至少两种的组合为IV级；
从图5(j)可以看出，对于所述印记基因Z13，印记基因缺失表达量小于10％、印记基因拷贝数异常表达量小于1％或印记基因总表达量小于25％中的任意一种或至少两种的组合为0级，印记基因缺失表达量为10-15％、印记基因拷贝数异常表达量为1-2％或印记基因总表达量为25-30％中的任意一种或至少两种的组合为I级，印记基因缺失表达量为15-25％、印记基因拷贝数异常表达量为2-3％或印记基因总表达量为30-45％中的任意一种或至少两种的组合为II级，印记基因缺失表达量为25-30％、印记基因拷贝数异常表达量为3-5％或印记基因总表达量为45-55％中的任意一种或至少两种的组合为III级，印记基因缺失表达量大于30％、印记基因拷贝数异常表达量大于5％或印记基因总表达量大于55％中的任意一种或至少两种的组合为IV级。
从这37例胃肿瘤的样本综合分析可以得出：
诊断肿瘤的良恶性程度分为良性肿瘤、胃癌潜能、早期胃癌、中期胃癌和晚期胃癌：
所述诊断胃肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16的印记基因缺失表达量和印记基因拷贝数异常表达量均小于I级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因缺失表达量为I级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因拷贝数异常表达量为I级中的任意一种情况，则为良性肿瘤；
所述诊断胃肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的至少2个印记基因的印记基因缺失表达量为I级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13的至少2个印记基因的印记基因拷贝数异常表达量为I级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、 Z11和Z13中的不超过1个印记基因的印记基因缺失表达量为I级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的不超过1个印记基因的印记基因拷贝数异常表达量为I级且印记基因Z16的印记基因缺失表达量或印记基因拷贝数异常表达量为I级或以上，或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中不超过1个印记基因的印记基因缺失表达量为II级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的不超过1个印记基因的印记基因拷贝数异常表达量为II级中的任意一种情况，则为胃癌潜能；
所述诊断胃肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的至少2个印记基因的印记基因缺失表达量为II级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的至少2个印记基因的印记基因拷贝数异常表达量为II级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中不超过1个印记基因的印记基因缺失表达量为III级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的不超过1个印记基因的印记基因拷贝数异常表达量为III级中的任意一种情况，则为早期胃癌；
所述诊断胃肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的至少2个印记基因的印记基因缺失表达量为III级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13的至少2个印记基因的印记基因拷贝数异常表达量为III级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中不超过1个印记基因的印记基因缺失表达量为IV级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的不超过1个印记基因的印记基因拷贝数异常表达量为IV级中的任意一种情况，则为中期胃癌；
所述诊断胃肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中至少2个印记基因的印记基因缺失表达量为IV级或印 记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中至少2个印记基因的印记基因拷贝数异常表达量为IV级，则为晚期胃癌。
实施例4 食道肿瘤的印记基因分析
获取食道肿瘤的组织细胞切片(10微米)，检测方法同实施例1。
从图6(a)-图6(e)可以看出，从0级到IV级的样本中，印记缺失(细胞核内有两个信号点)和拷贝数异常(细胞核内有三个或以上信号点)的细胞比例随恶性程度的增加而逐渐增加。
实施例5 食道的胃镜活检样本的印记基因分析
所述食道的胃镜活检样本是，在胃镜下取出食道的可疑病变组织，10％中性福尔马林溶液固定24h以上，其他检测方法同实施例1。
从图7(a)-图7(f)可以看出，Z1，Z3，Z4，Z5，Z6，Z8，Z10，Z11，Z13，Z16每个基因对食道癌的反应敏感性或者说对应于食道癌表达的印记缺失的强度和状态是不同的。
具体每个印记基因对食道癌的敏感度如图8(a)-图8(j)，从图8(a)-图8(c)可以看出，印记基因Z1的印记缺失在恶性潜能阶段开始出现，在早期食道癌中没有明显上升，在中期和晚期食道癌中逐渐上升到很高水平，印记基因Z1的拷贝数异常在恶性潜能到晚期食道癌阶段逐渐上升，印记基因Z1的表达量增加在食道癌发展过程中缓慢上升；印记基因Z11的印记缺失在恶性潜能阶段开始出现，在早期和中期食道癌阶段没有明显上升，到晚期食道癌阶段有比较明显的上升，印记基因Z11的拷贝数异常在恶性潜能阶段迅速上升，在早期到晚期食道癌的发展过程中上升速度逐渐减缓，印记基因Z11的表达量增加在恶性潜能到中期食道癌阶段都没有明显上升，但在晚期食道癌中有明显的上升；印记基因Z16的印记缺失、拷贝数异常和表达量增加在恶性潜能阶段开始出现， 在早期和中期食道癌阶段快速上升，到晚期食道癌阶段上升速度减缓；
从图8(d)-图8(j)可以看出，印记基因Z3的印记缺失、拷贝数异常和表达量增加在中期食道癌阶段开始上升，在晚期食道癌中继续上升，但水平仍然不高；印记基因Z4的印记缺失在中期食道癌阶段开始出现，但是在晚期食道癌阶段上升速度减缓，印记基因的拷贝数异常和表达量增加在中期食道癌阶段开始出现，在晚期食道癌中继续上升；印记基因Z5的印记缺失、拷贝数异常和表达量增加在中期食道癌阶段开始上升，在晚期食道癌中继续上升，但水平仍然不高；印记基因Z6的印记缺失在中期食道癌阶段开始出现，在晚期食道癌中加速上升，但水平仍然不高，印记基因Z6的拷贝数异常和表达量增加在恶性潜能阶段开始出现，随着食道癌的发展缓慢上升，到晚期食道癌阶段水平仍然不高；印记基因Z8的印记缺失和拷贝数异常在恶性潜能阶段开始出现，在早期和中期食道癌中快速上升，到晚期食道癌阶段上升速度减缓，印记基因Z8的表达量增加在恶性潜能阶段开始出现，但是在食道癌发展过程中没有明显上升；印记基因Z10的印记缺失在中期食道癌阶段开始出现，在晚期食道癌阶段没有继续上升，印记基因Z10的拷贝数异常在中期食道癌阶段开始出现，在晚期食道癌中继续上升，但水平不高，印记基因Z10的表达量增加在恶性潜能阶段开始出现，但是在食道癌发展过程中没有明显上升；印记基因Z13的印记缺失在早期食道癌中开始出现，在中期和晚期食道癌阶段逐渐上升，但水平不高，印记基因Z13的拷贝数异常和表达量增加在中期食道癌阶段快速上升，在晚期食道癌阶段上升速度减缓。
实施例6 57例食道肿瘤样本的印记基因分析
获取57例食道肿瘤病人的组织包括食道的胃镜活检样本(10微米)，检测方法同实施例1。
从图9(a)-图9(j)可以看出，57例食道肿瘤组织样本中Z1，Z3，Z4，Z5，Z6，Z8，Z10，Z11，Z13和Z16十个探针的印记缺失和拷贝数异常的比例呈现从低到高的分布，根据不同探针的分布趋势，我们计算得到了图中虚线所示的分级标准，可以将每个探针的印记缺失和拷贝数异常分别从低到高分成5个等级。
具体的分级如下：
从图9(a)可以看出，对于所述印记基因Z1，印记基因缺失表达量小于16％、印记基因拷贝数异常表达量小于2.5％或印记基因总表达量小于35％中的任意一种或至少两种的组合为0级，印记基因缺失表达量为16-20％、印记基因拷贝数异常表达量为2.5-4％或印记基因总表达量为35-40％中的任意一种或至少两种的组合为I级，印记基因缺失表达量为20-25％、印记基因拷贝数异常表达量为4-6.5％或印记基因总表达量为40-50％中的任意一种或至少两种的组合为II级，印记基因缺失表达量为25-30％、印记基因拷贝数异常表达量为6.5-8％或印记基因总表达量为50-65％中的任意一种或至少两种的组合为III级，印记基因缺失表达量大于30％、印记基因拷贝数异常表达量大于8％或印记基因总表达量大于65％中的任意一种或至少两种的组合为IV级；
从图9(b)可以看出，对于所述印记基因Z11，印记基因缺失表达量小于16％、印记基因拷贝数异常表达量小于1.5％或印记基因总表达量小于30％中的任意一种或至少两种的组合为0级，印记基因缺失表达量为16-20％、印记基因拷贝数异常表达量为1.5-2.5％或印记基因总表达量为30-40％中的任意一种或至少两种的组合为I级，印记基因缺失表达量为20-25％、印记基因拷贝数异常表达量为2.5-4％或印记基因总表达量为40-50％中的任意一种或至少两种的组合为II级，印记基因缺失表达量为25-30％、印记基因拷贝数异常表达量为4-6％或印 记基因总表达量为50-65％中的任意一种或至少两种的组合为III级，印记基因缺失表达量大于30％、印记基因拷贝数异常表达量大于6％或印记基因总表达量大于65％中的任意一种或至少两种的组合为IV级；
从图9(c)可以看出，对于所述印记基因Z16，印记基因缺失表达量小于10％、印记基因拷贝数异常表达量小于1％或印记基因总表达量小于25％中的任意一种或至少两种的组合为0级，印记基因缺失表达量为10-15％、印记基因拷贝数异常表达量为1-2％或印记基因总表达量为25-30％中的任意一种或至少两种的组合为I级，印记基因缺失表达量为15-25％、印记基因拷贝数异常表达量为2-4％或印记基因总表达量为30-40％中的任意一种或至少两种的组合为II级，印记基因缺失表达量为25-30％、印记基因拷贝数异常表达量为4-6％或印记基因总表达量为40-55％中的任意一种或至少两种的组合为III级，印记基因缺失表达量大于30％、印记基因拷贝数异常表达量大于6％或印记基因总表达量大于55％中的任意一种或至少两种的组合为IV级；
从图9(d)可以看出，对于所述印记基因Z3，印记基因缺失表达量小于10％、印记基因拷贝数异常表达量小于1％或印记基因总表达量小于25％中的任意一种或至少两种的组合为0级，印记基因缺失表达量为10-15％、印记基因拷贝数异常表达量为1-2％或印记基因总表达量为25-30％中的任意一种或至少两种的组合为I级，印记基因缺失表达量为15-25％、印记基因拷贝数异常表达量为2-3％或印记基因总表达量为30-45％中的任意一种或至少两种的组合为II级，印记基因缺失表达量为25-30％、印记基因拷贝数异常表达量为3-5％或印记基因总表达量为45-55％中的任意一种或至少两种的组合为III级，印记基因缺失表达量大于30％、印记基因拷贝数异常表达量大于5％或印记基因总表达量大于55％中的任意一种或至少两种的组合为IV级；
从图9(e)可以看出，对于所述印记基因Z4，印记基因缺失表达量小于16％、印记基因拷贝数异常表达量小于1.5％或印记基因总表达量小于30％中的任意一种或至少两种的组合为0级，印记基因缺失表达量为16-20％、印记基因拷贝数异常表达量为1.5-2.5％或印记基因总表达量为30-40％中的任意一种或至少两种的组合为I级，印记基因缺失表达量为20-25％、印记基因拷贝数异常表达量为2.5-4％或印记基因总表达量为40-50％中的任意一种或至少两种的组合为II级，印记基因缺失表达量为25-30％、印记基因拷贝数异常表达量为4-6％或印记基因总表达量为50-65％中的任意一种或至少两种的组合为III级，印记基因缺失表达量大于30％、印记基因拷贝数异常表达量大于6％或印记基因总表达量大于65％中的任意一种或至少两种的组合为IV级；
从图9(f)可以看出，对于所述印记基因Z5，印记基因缺失表达量小于16％、印记基因拷贝数异常表达量小于1.5％或印记基因总表达量小于30％中的任意一种或至少两种的组合为0级，印记基因缺失表达量为16-20％、印记基因拷贝数异常表达量为1.5-2.5％或印记基因总表达量为30-40％中的任意一种或至少两种的组合为I级，印记基因缺失表达量为20-25％、印记基因拷贝数异常表达量为2.5-4％或印记基因总表达量为40-50％中的任意一种或至少两种的组合为II级，印记基因缺失表达量为25-30％、印记基因拷贝数异常表达量为4-6％或印记基因总表达量为50-65％中的任意一种或至少两种的组合为III级，印记基因缺失表达量大于30％、印记基因拷贝数异常表达量大于6％或印记基因总表达量大于65％中的任意一种或至少两种的组合为IV级；
从图9(g)可以看出，对于所述印记基因Z6，印记基因缺失表达量小于16％、印记基因拷贝数异常表达量小于1.5％或印记基因总表达量小于30％中的任意一种或至少两种的组合为0级，印记基因缺失表达量为16-20％、印记基因拷贝数 异常表达量为1.5-2.5％或印记基因总表达量为30-40％中的任意一种或至少两种的组合为I级，印记基因缺失表达量为20-25％、印记基因拷贝数异常表达量为2.5-4％或印记基因总表达量为40-50％中的任意一种或至少两种的组合为II级，印记基因缺失表达量为25-30％、印记基因拷贝数异常表达量为4-6％或印记基因总表达量为50-65％中的任意一种或至少两种的组合为III级，印记基因缺失表达量大于30％、印记基因拷贝数异常表达量大于6％或印记基因总表达量大于65％中的任意一种或至少两种的组合为IV级；
从图9(h)可以看出，对于所述印记基因Z8，印记基因缺失表达量小于16％、印记基因拷贝数异常表达量小于2.5％或印记基因总表达量小于35％中的任意一种或至少两种的组合为0级，印记基因缺失表达量为16-20％、印记基因拷贝数异常表达量为2.5-4％或印记基因总表达量为35-40％中的任意一种或至少两种的组合为I级，印记基因缺失表达量为20-25％、印记基因拷贝数异常表达量为4-6.5％或印记基因总表达量为40-50％中的任意一种或至少两种的组合为II级，印记基因缺失表达量为25-30％、印记基因拷贝数异常表达量为6.5-8％或印记基因总表达量为50-65％中的任意一种或至少两种的组合为III级，印记基因缺失表达量大于30％、印记基因拷贝数异常表达量大于8％或印记基因总表达量大于65％中的任意一种或至少两种的组合为IV级；
从图9(i)可以看出，对于所述印记基因Z10，印记基因缺失表达量小于16％、印记基因拷贝数异常表达量小于1.5％或印记基因总表达量小于30％中的任意一种或至少两种的组合为0级，印记基因缺失表达量为16-20％、印记基因拷贝数异常表达量为1.5-2.5％或印记基因总表达量为30-40％中的任意一种或至少两种的组合为I级，印记基因缺失表达量为20-25％、印记基因拷贝数异常表达量为2.5-4％或印记基因总表达量为40-50％中的任意一种或至少两种的组合为 II级，印记基因缺失表达量为25-30％、印记基因拷贝数异常表达量为4-6％或印记基因总表达量为50-65％中的任意一种或至少两种的组合为III级，印记基因缺失表达量大于30％、印记基因拷贝数异常表达量大于6％或印记基因总表达量大于65％中的任意一种或至少两种的组合为IV级；
从图9(j)可以看出，对于所述印记基因Z13，印记基因缺失表达量小于10％、印记基因拷贝数异常表达量小于1％或印记基因总表达量小于25％中的任意一种或至少两种的组合为0级，印记基因缺失表达量为10-15％、印记基因拷贝数异常表达量为1-2％或印记基因总表达量为25-30％中的任意一种或至少两种的组合为I级，印记基因缺失表达量为15-25％、印记基因拷贝数异常表达量为2-3％或印记基因总表达量为30-45％中的任意一种或至少两种的组合为II级，印记基因缺失表达量为25-30％、印记基因拷贝数异常表达量为3-5％或印记基因总表达量为45-55％中的任意一种或至少两种的组合为III级，印记基因缺失表达量大于30％、印记基因拷贝数异常表达量大于5％或印记基因总表达量大于55％中的任意一种或至少两种的组合为IV级。
从这57例食道肿瘤的样本综合分析可以得出：
诊断食道肿瘤的良恶性程度分为良性肿瘤、食道癌潜能、早期食道癌、中期食道癌和晚期食道癌：
所述诊断食道肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16的印记基因缺失表达量和印记基因拷贝数异常表达量均小于I级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因缺失表达量为I级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因拷贝数异常表达量为I级中的任意一种情况，则为良性肿瘤；
所述诊断食道肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的至少2个印记基因的印记基因缺失表达量为I级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16的至少2个印记基因的印记基因拷贝数异常表达量为I级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中不超过1个印记基因的印记基因缺失表达量为II级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因拷贝数异常表达量为II级中的任意一种情况，则为食道癌潜能；
所述诊断食道肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的至少2个印记基因的印记基因缺失表达量为II级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16的至少2个印记基因的印记基因拷贝数异常表达量为II级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中不超过1个印记基因的印记基因缺失表达量为III级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因拷贝数异常表达量为III级中的任意一种情况，则为早期食道癌；
所述诊断食道肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的至少2个印记基因的印记基因缺失表达量为III级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16的至少2个印记基因的印记基因拷贝数异常表达量为III级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中不超过1个印记基因的印记基因缺失表达量为IV级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因拷贝数异常表达量为IV级中的任意一种情 况，则为中期食道癌；
所述诊断食道肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中至少2个印记基因的印记基因缺失表达量为IV级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中至少2个印记基因的印记基因拷贝数异常表达量为IV级，则为晚期食道癌。
综上所述，本公开所述检测模型和系统，以直观的方法表现了印记缺失在食道和胃肿瘤病人的样本上的表现，通过对印记基因原位标记的方法，客观，直观，早期，精确地检测出印记(迹)基因的变化，并可以提供量化的模型，为食道和胃肿瘤的诊断做出巨大贡献。
申请人声明，本公开通过上述实施例来说明本公开的详细方法，但本公开并不局限于上述详细方法，即不意味着本公开必须依赖上述详细方法才能实施。所属技术领域的技术人员应该明了，对本公开的任何改进，对本公开产品各原料的等效替换及辅助成分的添加、具体方式的选择等，均落在本公开的保护范围和公开范围之内。

Claims (21)

  1. 一种用于食道肿瘤和/或胃肿瘤的印记基因分级模型，其通过计算印记基因缺失表达量、印记基因拷贝数异常表达量和印记基因的总表达量在食道肿瘤和/或胃肿瘤中的变化对印记基因的表达状态进行分级；
    其中，所述印记基因为Z1、Z11或Z16中的任意一个或至少两个的组合，所述印记基因Z1为Gnas，所述印记基因Z11为Grb10，所述印记基因Z16为Snrpn/Snurf。
  2. 根据权利要求1所述的印记基因分级模型，其中，所述模型计算印记基因的方法如下：
    计算Z1、Z11或Z16中的任意一个，优选为Z1或Z11中的任意一个。
  3. 根据权利要求1或2所述的印记基因分级模型，其中，用于检测食道肿瘤时，所述模型计算的印记基因为Z1。
  4. 根据权利要求1-3中任一项所述的印记基因分级模型，其中，用于检测胃肿瘤时，所述模型计算的印记基因为Z11。
  5. 根据权利要求1-4中任一项所述的印记基因分级模型，其中，所述模型计算印记基因的方法为：计算Z1、Z11或Z16中的任意两个印记基因的组合。
  6. 根据权利要求1-5中任一项所述的印记基因分级模型，其中，所述印记基因还包括Z3、Z4、Z5、Z6、Z8、Z10或Z13中的任意一个或至少两个的组合；其中，所述印记基因Z3为Peg10，所述印记基因Z4为Igf2r，所述印记基因Z5为Mest，所述印记基因Z6为Plagl1，所述印记基因Z8为Dcn，所述印记基因Z10为Gatm，所述印记基因Z13为Sgce。
  7. 根据权利要求1-6中任一项所述的印记基因分级模型，其中，所述模型计算印记基因的方法为：计算印记基因的组合，计算Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16的十个印记基因的组合。
  8. 根据权利要求1-7中任一项所述的印记基因分级模型，其中，所述计算印记基因的总表达量、印记基因缺失表达量和印记基因拷贝数异常表达量的公式如下：
    总表达量＝(b+c+d)/(a+b+c+d)×100％；
    正常印记基因总表达量＝b/(b+c+d)×100％；
    印记基因缺失基因表达量＝c/(b+c+d)×100％；
    印记基因拷贝数异常的基因表达量＝d/(b+c+d)×100％；
    其中，所述a为将细胞进行苏木素染色后，细胞核内不存在标记，印记基因没有表达的细胞核；所述b为将细胞进行苏木素染色后，细胞核内存在一个红色/棕色标记，印记基因存在的细胞核；所述c为将细胞进行苏木素染色后，细胞核内存在两个红色/棕色标记，印记基因缺失的细胞核；所述d为将细胞进行苏木素染色后，细胞核内存在两个以上红色/棕色标记，印记基因拷贝数异常的细胞核。
  9. 根据权利要求7所述的印记基因分级模型，其中，所述印记基因缺失表达量、印记基因拷贝数异常表达量和印记基因总表达量分成五个不同的等级；
    所述五个不同的等级为针对Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16的十个印记基因的印记基因缺失表达量、印记基因拷贝数异常表达量和印记基因总表达量分别进行划分的五个不同的等级；
    所述针对Z1的印记基因缺失表达量、印记基因拷贝数异常表达量和总表达量划分的五个不同的等级为：
    0级：所述印记基因Z1的印记基因缺失表达量小于16％、所述印记基因Z1的印记基因拷贝数异常表达量小于2.5％或所述印记基因Z1的总表达量小于35％中的任意一种或至少两种的组合；
    I级：所述印记基因Z1的印记基因缺失表达量为16-20％、所述印记基因Z1的印记基因拷贝数异常表达量为2.5-4％或所述印记基因Z1的总表达量为35-40％中的任意一种或至少两种的组合；
    II级：所述印记基因Z1的印记基因缺失表达量为20-25％、所述印记基因Z1的印记基因拷贝数异常表达量为4-6.5％或所述印记基因Z1的总表达量为40-50％中的任意一种或至少两种的组合；
    III级：所述印记基因Z1的印记基因缺失表达量为25-30％、所述印记基因Z1的印记基因拷贝数异常表达量为6.5-8％或所述印记基因Z1的总表达量为50-65％中的任意一种或至少两种的组合；
    IV级：所述印记基因Z1的印记基因缺失表达量大于30％、所述印记基因Z1的印记基因拷贝数异常表达量大于8％或所述印记基因Z1的总表达量大于65％中的任意一种或至少两种的组合；
    所述针对Z3和Z13的印记基因缺失表达量、印记基因拷贝数异常表达量和总表达量划分的五个不同的等级为：
    0级：所述印记基因Z3和Z13的印记基因缺失表达量小于10％、所述印记基因Z3和Z13的印记基因拷贝数异常表达量小于1％或所述印记基因Z3和Z13的总表达量小于25％中的任意一种或至少两种的组合；
    I级：所述印记基因Z3和Z13的印记基因缺失表达量为10-15％、所述印记基因Z3和Z13的印记基因拷贝数异常表达量为1-2％或所述印记基因Z3和Z13的总表达量为25-30％中的任意一种或至少两种的组合；
    II级：所述印记基因Z3和Z13的印记基因缺失表达量为15-25％、所述印记基因Z3和Z13的印记基因拷贝数异常表达量为2-3％或所述印记基因Z3和Z13的总表达量为30-45％中的任意一种或至少两种的组合；
    III级：所述印记基因Z3和Z13的印记基因缺失表达量为25-30％、所述印记基因Z3和Z13的印记基因拷贝数异常表达量为3-5％或所述印记基因Z3和Z13的总表达量为45-55％中的任意一种或至少两种的组合；
    IV级：所述印记基因Z3和Z13的印记基因缺失表达量大于30％、所述印记基因Z3和Z13的印记基因拷贝数异常表达量大于5％或所述印记基因Z3和Z13的总表达量大于55％中的任意一种或至少两种的组合；
    所述针对Z4、Z5、Z6、Z10和Z11的印记基因缺失表达量、印记基因拷贝数异常表达量和总表达量划分的五个不同的等级为：
    0级：所述印记基因Z4、Z5、Z6、Z10和Z11的印记基因缺失表达量小于16％、所述印记基因Z4、Z5、Z6、Z10和Z11的印记基因拷贝数异常表达量小于1.5％或所述印记基因Z4、Z5、Z6、Z10和Z11的总表达量小于30％中的任意一种或至少两种的组合；
    I级：所述印记基因Z4、Z5、Z6、Z10和Z11的印记基因缺失表达量为16-20％、所述印记基因Z4、Z5、Z6、Z10和Z11的印记基因拷贝数异常表达量为1.5-2.5％或所述印记基因Z4、Z5、Z6、Z10和Z11的总表达量为30-40％中的任意一种或至少两种的组合；
    II级：所述印记基因Z4、Z5、Z6、Z10和Z11的印记基因缺失表达量为20-25％、所述印记基因Z4、Z5、Z6、Z10和Z11的印记基因拷贝数异常表达量为2.5-4％或所述印记基因Z4、Z5、Z6、Z10和Z11的总表达量为40-50％中的任意一种或至少两种的组合；
    III级：所述印记基因Z4、Z5、Z6、Z10和Z11的印记基因缺失表达量为25-30％、所述印记基因Z4、Z5、Z6、Z10和Z11的印记基因拷贝数异常表达量为4-6％或所述印记基因Z4、Z5、Z6、Z10和Z11的总表达量为50-65％中的任 意一种或至少两种的组合；
    IV级：所述印记基因Z4、Z5、Z6、Z10和Z11的印记基因缺失表达量大于30％、所述印记基因Z4、Z5、Z6、Z10和Z11的印记基因拷贝数异常表达量大于6％或所述印记基因Z4、Z5、Z6、Z10和Z11的总表达量大于65％中的任意一种或至少两种的组合；
    用于检测食道肿瘤时，所述针对Z16的印记基因缺失表达量、印记基因拷贝数异常表达量和总表达量划分的五个不同的等级为：
    0级：所述印记基因Z16的印记基因缺失表达量小于10％、所述印记基因Z16的印记基因拷贝数异常表达量小于1％或所述印记基因Z16的总表达量小于25％中的任意一种或至少两种的组合；
    I级：所述印记基因Z16的印记基因缺失表达量为10-15％、所述印记基因Z16的印记基因拷贝数异常表达量为1-2％或所述印记基因Z16的总表达量为25-30％中的任意一种或至少两种的组合；
    II级：所述印记基因Z16的印记基因缺失表达量为15-25％、所述印记基因Z16的印记基因拷贝数异常表达量为2-4％或所述印记基因Z16的总表达量为30-40％中的任意一种或至少两种的组合；
    III级：所述印记基因Z16的印记基因缺失表达量为25-30％、所述印记基因Z16的印记基因拷贝数异常表达量为4-6％或所述印记基因Z16的总表达量为40-55％中的任意一种或至少两种的组合；
    IV级：所述印记基因Z16的印记基因缺失表达量大于30％、所述印记基因Z16的印记基因拷贝数异常表达量大于6％或所述印记基因Z16的总表达量大于55％中的任意一种或至少两种的组合；
    用于检测胃肿瘤时，所述针对Z16的印记基因缺失表达量、印记基因拷贝 数异常表达量和总表达量划分的五个不同的等级为：
    0级：所述印记基因Z16的印记基因缺失表达量小于15％、所述印记基因Z16的印记基因拷贝数异常表达量小于1.5％或所述印记基因Z16的总表达量小于30％中的任意一种或至少两种的组合；
    I级：所述印记基因Z16的印记基因缺失表达量为15-20％、所述印记基因Z16的印记基因拷贝数异常表达量为1.5-4％或所述印记基因Z16的总表达量为30-40％中的任意一种或至少两种的组合；
    II级：所述印记基因Z16的印记基因缺失表达量为20-25％、所述印记基因Z16的印记基因拷贝数异常表达量为4-7％或所述印记基因Z16的总表达量为40-50％中的任意一种或至少两种的组合；
    III级：所述印记基因Z16的印记基因缺失表达量为25-30％、所述印记基因Z16的印记基因拷贝数异常表达量为7-10％或所述印记基因Z16的总表达量为50-60％中的任意一种或至少两种的组合；
    IV级：所述印记基因Z16的印记基因缺失表达量大于30％、所述印记基因Z16的印记基因拷贝数异常表达量大于10％或所述印记基因Z16的总表达量大于60％中的任意一种或至少两种的组合；
    用于检测食道肿瘤时，所述针对Z8的印记基因缺失表达量、印记基因拷贝数异常表达量和总表达量划分的五个不同的等级为：
    0级：所述印记基因Z8的印记基因缺失表达量小于16％、所述印记基因Z8的印记基因拷贝数异常表达量小于2.5％或所述印记基因Z8的总表达量小于35％中的任意一种或至少两种的组合；
    I级：所述印记基因Z8的印记基因缺失表达量为16-20％、所述印记基因Z8的印记基因拷贝数异常表达量为2.5-4％或所述印记基因Z8的总表达量为 35-40％中的任意一种或至少两种的组合；
    II级：所述印记基因Z8的印记基因缺失表达量为20-25％、所述印记基因Z8的印记基因拷贝数异常表达量为4-6.5％或所述印记基因Z8的总表达量为40-50％中的任意一种或至少两种的组合；
    III级：所述印记基因Z8的印记基因缺失表达量为25-30％、所述印记基因Z8的印记基因拷贝数异常表达量为6.5-8％或所述印记基因Z8的总表达量为50-65％中的任意一种或至少两种的组合；
    IV级：所述印记基因Z8的印记基因缺失表达量大于30％、所述印记基因Z8的印记基因拷贝数异常表达量大于8％或所述印记基因Z8的总表达量大于65％中的任意一种或至少两种的组合；
    用于检测胃肿瘤时，所述针对Z8的印记基因缺失表达量、印记基因拷贝数异常表达量和总表达量划分的五个不同的等级为：
    0级：所述印记基因Z8的印记基因缺失表达量小于10％、所述印记基因Z8的印记基因拷贝数异常表达量小于1％或所述印记基因Z8的总表达量小于25％中的任意一种或至少两种的组合；
    I级：所述印记基因Z8的印记基因缺失表达量为10-15％、所述印记基因Z8的印记基因拷贝数异常表达量为1-2％或所述印记基因Z8的总表达量为25-30％中的任意一种或至少两种的组合；
    II级：所述印记基因Z8的印记基因缺失表达量为15-25％、所述印记基因Z8的印记基因拷贝数异常表达量为2-3％或所述印记基因Z8的总表达量为30-45％中的任意一种或至少两种的组合；
    III级：所述印记基因Z8的印记基因缺失表达量为25-30％、所述印记基因Z8的印记基因拷贝数异常表达量为3-5％或所述印记基因Z8的总表达量为 45-55％中的任意一种或至少两种的组合；
    IV级：所述印记基因Z8的印记基因缺失表达量大于30％、所述印记基因Z8的印记基因拷贝数异常表达量大于5％或所述印记基因Z8的总表达量大于55％中的任意一种或至少两种的组合。
  10. 一种用于检测食道肿瘤和/或胃肿瘤良恶性程度的装置，采用如权利要求1-9中任一项所述的印记基因分级模型，包括如下单元：
    (1)取样单元：获取待测样本；
    (2)探针设计单元：根据印记基因序列设计特异性引物；
    (3)检测单元：将步骤(2)的探针与待测样本进行原位杂交；
    (4)分析单元：显微镜成像分析印记基因的表达情况；
    其中，所述分析单元通过计算印记基因缺失表达量和印记基因拷贝数异常表达量，通过权利要求1-9中任一项所述的印记基因分级模型，从而通过印记基因缺失表达量和印记基因拷贝数异常表达量的等级来诊断食道肿瘤和/或胃肿瘤的良恶性程度。
  11. 一种用于检测食道肿瘤和/或胃肿瘤良恶性程度的方法，采用如权利要求1-9中任一项所述的印记基因分级模型，包括如下步骤：
    (1)获取待测样本；
    (2)根据印记基因序列设计特异性引物；
    (3)将步骤(2)的探针与待测样本进行原位杂交；
    (4)显微镜成像分析印记基因的表达情况，从而诊断乳腺肿瘤的良恶性程度；
    其中，所述分析单元通过计算印记基因缺失表达量和印记基因拷贝数异常表达量，通过权利要求1-9中任一项所述的印记基因分级模型，从而通过印记 基因缺失表达量和印记基因拷贝数异常表达量的等级来诊断食道肿瘤和/或胃肿瘤的良恶性程度。
  12. 根据权利要求11所述的方法，其中，步骤(1)所述的待测样本来自于人的组织和/或细胞。
  13. 根据权利要求11或12所述的方法，其中，所述待测样本为组织的石蜡切片、食道黏膜脱落细胞或胃镜活检样本，优选为食道黏膜脱落细胞和/或胃镜活检样本。
  14. 根据权利要求11-13中任一项所述的方法，其中，所述原位杂交采用RNAscope原位杂交方法。
  15. 根据权利要求11-14中任一项所述的方法，其中，所述RNAscope原位杂交方法使用单通道或多通道的呈色试剂盒或者单通道或多通道的荧光试剂盒，优选为单通道红色/棕色呈色试剂盒或多通道的荧光试剂盒。
  16. 根据权利要求11-15中任一项所述的方法，其中，所述诊断食道肿瘤的良恶性程度分为良性肿瘤、食道癌潜能、早期食道癌、中期食道癌和晚期食道癌。
  17. 根据权利要求11-16中任一项所述的方法，其中，所述判断食道肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16的印记基因缺失表达量和印记基因拷贝数异常表达量均小于I级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因缺失表达量为I级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因拷贝数异常表达量为I级中的任意一种情况，则为良性肿瘤；
    所述诊断食道肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、 Z6、Z8、Z10、Z11、Z13和Z16中的至少2个印记基因的印记基因缺失表达量为I级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16的至少2个印记基因的印记基因拷贝数异常表达量为I级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中不超过1个印记基因的印记基因缺失表达量为II级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因拷贝数异常表达量为II级中的任意一种情况，则为食道癌潜能；
    所述诊断食道肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的至少2个印记基因的印记基因缺失表达量为II级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16的至少2个印记基因的印记基因拷贝数异常表达量为II级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中不超过1个印记基因的印记基因缺失表达量为III级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因拷贝数异常表达量为III级中的任意一种情况，则为早期食道癌；
    所述诊断食道肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的至少2个印记基因的印记基因缺失表达量为III级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16的至少2个印记基因的印记基因拷贝数异常表达量为III级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中不超过1个印记基因的印记基因缺失表达量为IV级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因拷贝数异常表达量为IV级中的任意一种情况，则为中期食道癌；
    所述诊断食道肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中至少2个印记基因的印记基因缺失表达量为IV级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中至少2个印记基因的印记基因拷贝数异常表达量为IV级，则为晚期食道癌；
    所述诊断胃肿瘤的良恶性程度分为良性胃肿瘤、胃癌潜能、早期胃癌、中期胃癌和晚期胃癌；
    所述诊断胃肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16的印记基因缺失表达量和印记基因拷贝数异常表达量均小于I级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因缺失表达量为I级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因拷贝数异常表达量为I级中的任意一种情况，则为良性肿瘤；
    所述诊断胃肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的至少2个印记基因的印记基因缺失表达量为I级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13的至少2个印记基因的印记基因拷贝数异常表达量为I级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的不超过1个印记基因的印记基因缺失表达量为I级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的不超过1个印记基因的印记基因拷贝数异常表达量为I级且印记基因Z16的印记基因缺失表达量或印记基因拷贝数异常表达量为I级或以上，或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中不超过1个印记基因的印记基因缺失表达量为II级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的不超过1个印记基因的印记基因拷贝数异常表达量为II级中的任意一种情况，则为胃癌潜能；
    所述诊断胃肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的至少2个印记基因的印记基因缺失表达量为II级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的至少2个印记基因的印记基因拷贝数异常表达量为II级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中不超过1个印记基因的印记基因缺失表达量为III级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的不超过1个印记基因的印记基因拷贝数异常表达量为III级中的任意一种情况，则为早期胃癌；
    所述诊断胃肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的至少2个印记基因的印记基因缺失表达量为III级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13的至少2个印记基因的印记基因拷贝数异常表达量为III级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中不超过1个印记基因的印记基因缺失表达量为IV级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的不超过1个印记基因的印记基因拷贝数异常表达量为IV级中的任意一种情况，则为中期胃癌；
    所述诊断胃肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中至少2个印记基因的印记基因缺失表达量为IV级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中至少2个印记基因的印记基因拷贝数异常表达量为IV级，则为晚期胃癌。
  18. 一种如权利要求1-9中任一项所述的印记基因分级模型和/或如权利要求10所述的装置用于食道肿瘤和/或胃肿瘤检测。
  19. 一种如权利要求1-9中任一项所述的印记基因分级模型和/或如权利要求10所述的装置用于制备治疗食道肿瘤和/或胃肿瘤的药物或器械的用途。
  20. 根据权利要求19所述的用途，其中，诊断食道肿瘤的良恶性程度分为 良性肿瘤、食道癌潜能、早期食道癌、中期食道癌和晚期食道癌；
    所述诊断食道肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16的印记基因缺失表达量和印记基因拷贝数异常表达量均小于I级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因缺失表达量为I级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因拷贝数异常表达量为I级中的任意一种情况，则为良性肿瘤；
    所述诊断食道肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的至少2个印记基因的印记基因缺失表达量为I级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16的至少2个印记基因的印记基因拷贝数异常表达量为I级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中不超过1个印记基因的印记基因缺失表达量为II级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因拷贝数异常表达量为II级中的任意一种情况，则为食道癌潜能；
    所述诊断食道肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的至少2个印记基因的印记基因缺失表达量为II级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16的至少2个印记基因的印记基因拷贝数异常表达量为II级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中不超过1个印记基因的印记基因缺失表达量为III级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因拷贝数异常表达量为III级中的任意一种情况，则为早期食道癌；
    所述诊断食道肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的至少2个印记基因的印记基因缺失表达量为III级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16的至少2个印记基因的印记基因拷贝数异常表达量为III级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中不超过1个印记基因的印记基因缺失表达量为IV级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因拷贝数异常表达量为IV级中的任意一种情况，则为中期食道癌；
    所述诊断食道肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中至少2个印记基因的印记基因缺失表达量为IV级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中至少2个印记基因的印记基因拷贝数异常表达量为IV级，则为晚期食道癌；
    所述诊断胃肿瘤的良恶性程度分为良性胃肿瘤、胃癌潜能、早期胃癌、中期胃癌和晚期胃癌；
    所述诊断胃肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16的印记基因缺失表达量和印记基因拷贝数异常表达量均小于I级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因缺失表达量为I级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11、Z13和Z16中的不超过1个印记基因的印记基因拷贝数异常表达量为I级中的任意一种情况，则为良性肿瘤；
    所述诊断胃肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的至少2个印记基因的印记基因缺失表达量为I级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13的至少2个印记基因的印 记基因拷贝数异常表达量为I级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的不超过1个印记基因的印记基因缺失表达量为I级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的不超过1个印记基因的印记基因拷贝数异常表达量为I级且印记基因Z16的印记基因缺失表达量或印记基因拷贝数异常表达量为I级或以上，或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中不超过1个印记基因的印记基因缺失表达量为II级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的不超过1个印记基因的印记基因拷贝数异常表达量为II级中的任意一种情况，则为胃癌潜能；
    所述诊断胃肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的至少2个印记基因的印记基因缺失表达量为II级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的至少2个印记基因的印记基因拷贝数异常表达量为II级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中不超过1个印记基因的印记基因缺失表达量为III级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的不超过1个印记基因的印记基因拷贝数异常表达量为III级中的任意一种情况，则为早期胃癌；
    所述诊断胃肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的至少2个印记基因的印记基因缺失表达量为III级，印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13的至少2个印记基因的印记基因拷贝数异常表达量为III级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中不超过1个印记基因的印记基因缺失表达量为IV级且印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中的不超过1个印记基因的印记基因拷贝数异常表达量为IV级中的任意一种情况，则为中期胃癌；
    所述诊断胃肿瘤的良恶性程度的结果为印记基因Z1、Z3、Z4、Z5、Z6、 Z8、Z10、Z11和Z13中至少2个印记基因的印记基因缺失表达量为IV级或印记基因Z1、Z3、Z4、Z5、Z6、Z8、Z10、Z11和Z13中至少2个印记基因的印记基因拷贝数异常表达量为IV级，则为晚期胃癌；
  21. 一种晚期胃癌诊断的标志物，其中，所述标志物为印记基因Z1和印记基因Z11；
    其中，所述印记基因Z1为Gnas，所述印记基因Z11为Grb10。
    根据权利要求21所述的标志物，其中，所述晚期胃癌为胃癌的TNM等级大于T2N0M0的胃癌。
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