WO2021246780A1 - Composition for diagnosis of cancer metastasis and recurrence - Google Patents

Abstract

The present invention relates to a biomarker including at least one protein selected from the group consisting of CD110, CDCP1, ABCA1, and IDOL or a gene coding therefor, a composition and a kit for diagnosis of cancer metastasis or recurrence, and a method for providing information for diagnosis of cancer metastasis or recurrence, wherein the composition and the kit each comprises an agent for measuring a level of at least one protein selected from the group consisting of CD110, CDCP1, ABCA1, and IDOL, or an expression level of a gene coding therefor. The method for diagnosis of cancer metastasis and recurrence by using the composition for diagnosis of cancer metastasis or recurrence, the composition comprising an agent for measuring a level of at least one protein selected from the group consisting of CD110, CDCP1, ABCA1, and IDOL or an expression level of a gene coding therefor according to the present disclosure can accurately diagnose cancer metastasis or recurrence in a non-invasive manner and as such, can diagnose cancer metastasis or recurrence more stably and easily than invasive methods.

Description

Composition for diagnosing cancer metastasis and recurrence

The present invention relates to a biomarker comprising one or more proteins selected from the group consisting of CD110, CDCP1, ABCA1, IDOL, or a gene encoding the same, and the level of any one or more proteins selected from the group consisting of CD110, CDCP1, ABCA1, IDOL, or To a composition and kit for diagnosing cancer metastasis or recurrence, and a method for providing information for diagnosing cancer metastasis or recurrence, which include an agent for measuring the expression level of a gene encoding the same.

Cancer metastasis refers to a process in which cancer cells are transferred from an initial site of occurrence to another site and grow again through interaction of cancer cells in that other site. Most cancer cells die on their own after leaving the primary site, where the cancer first occurred, as they no longer receive survival signals from surrounding cells or tissues, but some cancer cells survive and ultimately cause metastasis do. In particular, colorectal cancer can be expected to be cured if it is detected at an early stage, but as the detection time is delayed, metastasis occurs to places that are difficult to resection, such as the lungs, liver, lymph nodes, or peritoneum. In most cases, colon cancer has no symptoms in its early stages, and when symptoms appear, it is often quite advanced. Therefore, it is important to diagnose it early, but since there are often no specific abnormal signs, it is difficult to diagnose colon cancer metastasis (KR 10-2006999 B1).

In addition, since the risk of recurrence remains after chemical or surgical chemotherapy, early diagnosis of recurrence is important.

Invasive biopsy is mainly used as a method of diagnosing cancer, but surgical operation is required to collect cancer tissue from the lesion site, causing pain to the patient, and difficult tissue acquisition depending on the presence or absence of cancer metastasis. , repeated biopsy may be required, which puts a great burden on the patient. Accordingly, a method of using a blood sample, which is a non-invasive method, has been tried as a method of diagnosing cancer, but the accuracy is low, and thus it can be used only as a reference test.

[Prior art literature]

[Patent Literature]

(Patent Document 1) KR 10-2006999 B1

Accordingly, the present inventors have studied to develop a non-invasive, early diagnosis, and high-accuracy biomarker for diagnosing cancer metastasis or recurrence. As a result, CD110 and CDCP1, which have not been previously reported to be detected in blood, are detected in cancer metastasis or recurrence. To complete the present invention by confirming for the first time that the expression level is increased in a blood sample.

Accordingly, it is an object of the present invention to provide a biomarker for diagnosing cancer metastasis or recurrence in a non-invasive method and a composition for diagnosing cancer metastasis or recurrence.

Another object of the present invention is to provide an information providing method for diagnosing metastasis or recurrence of cancer using the biomarker.

In order to achieve the above object, one aspect of the present invention provides a blood sample collected from an individual comprising an agent for measuring the level of one or more proteins selected from the group consisting of CD110 and CDCP1 or the expression level of a gene encoding the same. Provided is a composition for diagnosing cancer metastasis or recurrence.

Another aspect of the present invention provides a kit for diagnosing cancer metastasis or recurrence using a blood sample collected from an individual comprising the composition.

In another aspect of the present invention, measuring the level of any one or more proteins selected from the group consisting of CD110 and CDCP1 or the expression level of a gene encoding the same in a blood sample isolated from an individual having cancer; And it provides an information providing method for diagnosing cancer metastasis, comprising determining a cancer metastasis state when the measured protein level or the expression level of a gene encoding the same is higher than that of an individual without cancer metastasis.

In another aspect of the present invention, measuring the level of any one or more proteins selected from the group consisting of CD110 and CDCP1 or the expression level of a gene encoding the same in a blood sample isolated from a subject undergoing tumor treatment; And it provides an information providing method for diagnosing cancer recurrence, comprising the step of judging a cancer recurrence state when the measured level of the protein or the expression level of the gene encoding the same is higher than that of an individual without cancer recurrence.

Another aspect of the present invention is for diagnosing cancer metastasis or recurrence using a blood sample collected from an individual comprising an agent for measuring the level of one or more proteins selected from the group consisting of ABCA1 and IDOL or the expression level of a gene encoding the same A composition is provided.

Another aspect of the present invention provides a kit for diagnosing cancer metastasis or recurrence using a blood sample collected from an individual comprising the composition.

In another aspect of the present invention, measuring the level of one or more proteins selected from the group consisting of ABCA1 and IDOL or the expression level of a gene encoding the same in a blood sample isolated from an individual having cancer; And when the measured level of ABCA1 protein or the expression level of a gene encoding the same is low compared to a non-cancerous individual, it is determined as a cancer metastasis state, and the measured level of IDOL protein or expression of a gene encoding the same It provides a method of providing information for diagnosing cancer metastasis, comprising determining a cancer metastasis state when the level is higher than that of an individual who has not metastasized.

In another aspect of the present invention, measuring the level of one or more proteins selected from the group consisting of ABCA1 and IDOL or the expression level of a gene encoding the same in a blood sample isolated from a subject undergoing tumor treatment; and a case where the measured level of ABCA1 protein or the expression level of a gene encoding the same is low compared to a non-cancerous individual as a cancer recurrence state, and the measured level of IDOL protein or expression of a gene encoding the same It provides a method for providing information for diagnosing cancer recurrence, comprising determining a cancer recurrence state when the level is higher than that of an individual with no recurrence of cancer.

Another aspect of the present invention provides a composition using a blood sample collected from an individual comprising an agent for measuring the level of one or more proteins selected from the group consisting of CD110 and CDCP1 or the expression level of a gene encoding the same. Provided is a use for diagnosing relapse.

Another aspect of the present invention provides a composition using a blood sample collected from an individual comprising an agent for measuring the level of one or more proteins selected from the group consisting of CD110 and CDCP1 or the expression level of a gene encoding the same. Provided is a use for manufacturing a medicament for diagnosis of recurrence.

Another aspect of the present invention provides the use of a composition comprising an agent for measuring the level of one or more proteins selected from the group consisting of ABCA1 and IDOL or the expression level of a gene encoding the same for diagnosing cancer metastasis or recurrence do.

Another aspect of the present invention is the use of a composition comprising an agent for measuring the level of one or more proteins selected from the group consisting of ABCA1 and IDOL or the expression level of a gene encoding the same for preparing a medicament for diagnosing cancer metastasis or recurrence provides

Cancer metastasis and recurrence using a composition for diagnosing cancer metastasis or recurrence comprising an agent for measuring the level of one or more proteins selected from the group consisting of CD110, CDCP1, ABCA1 and IDOL according to the present invention or the expression level of a gene encoding the same Since the method for diagnosis can accurately diagnose cancer metastasis or recurrence in a non-invasive method, it is possible to diagnose cancer metastasis or recurrence more safely and easily than an invasive method.

In addition, the composition for diagnosing cancer metastasis or recurrence according to the present invention may further include an agent for measuring the level of one or more proteins selected from the group consisting of EpCam and SQLE or the expression level of a gene encoding the same, in this case , it can diagnose cancer metastasis or recurrence with higher accuracy.

1 is a view of the proliferation of colorectal cancer cell lines (HT29 and HCT116) according to the treatment of cholesterol using a non-adhesive plate or the degree of inhibition of squalene epoxidase (SQLE) expression using light microscopy and MTT analysis. It is the result.

2 shows the gene expression levels of cancer cell markers (CD110 and CDCP1), cholesterol influx related protein (IDOL) and cholesterol release related protein (ABCA1) in colorectal cancer cell lines (HCT116 and HT29) according to the treatment of cholesterol using non-adherent plates. is the result confirmed using Quantitative PCR.

Figure 3a is the result of confirming the expression levels of CD45, cancer stem cell marker (EpCam), cancer cell markers (CD110 and CDCP1) and SQLE in colorectal cancer cell line (HCT116) according to the treatment of cholesterol using flow cytometry analysis. , Figure 3b shows the expression levels of CD45, cancer stem cell markers (EpCam), cancer cell markers (CD110 and CDCP1) and SQLE in colorectal cancer cell line (HT29) according to the treatment of cholesterol using flow cytometry analysis. to be.

4A is a schematic diagram illustrating a method for preparing an animal model of cancer metastasis.

Figure 4b is a cancer metastasis animal model mouse (HCT116 p53 ^-/- , HT29 or HCT116) by adjusting the diet for the measurement results of serum cholesterol at 5 weeks, 9 weeks, and 25 to 26 weeks from the day starting breeding. it has been shown

4c and 4d are cancer metastasis animal model mice (HT29) breeding for 30 days, 60 days, 90 days and 120 days using in-vivo bioluminescence imaging (BLi) according to the treatment of cholesterol and the degree of inhibition of SQLE expression These are the results of photographing the degree of cancer metastasis (FIG. 4c) and digitizing it (FIG. 4D).

4e and 4f show the treatment of cholesterol and the expression inhibition of SQLE using in-vivo bioluminescence imaging (BLi) while breeding cancer metastasis animal model mice (HCT116) for 30 days, 60 days, 90 days and 120 days. These are the results of photographing the degree of cancer metastasis (FIG. 4e) and digitization (FIG. 4F).

Figure 5a is a result of analyzing the level of colorectal cancer cell marker (CDCP1) in lung tissue according to the degree of inhibition of cholesterol treatment and SQLE expression in a cancer metastasis mouse model (HT29) using a confocal fluorescence microscope.

Figure 5b is a result of analyzing the level of colorectal cancer cell marker (CDCP1) in lung tissue according to the degree of inhibition of cholesterol treatment and SQLE expression in a cancer metastasis mouse model (HCT116) using a confocal fluorescence microscope.

Figures 6a and 6c flow cytometry analysis of the levels of cancer stem cell markers (EpCam), cancer cell markers (CD110 and CDCP1) and SQLE according to the level of inhibition of the expression of SQLE and the treatment of cholesterol in blood samples from cancer metastasis animal model mice (HT29). It is the result of analysis (FIG. 6a) and numerical value (FIG. 6c) using

6b and 6d are flow cytometry analysis of the levels of cancer stem cell markers (EpCam), cancer cell markers (CD110 and CDCP1) and SQLE according to the degree of cholesterol treatment and SQLE expression inhibition in blood samples from cancer metastasis animal model mice (HCT116). It is the result of analysis (FIG. 6b) and numerical value (FIG. 6D) using

Hereinafter, the present invention will be described in detail.

One aspect of the present invention provides a composition for diagnosing cancer metastasis or recurrence using a blood sample collected from an individual comprising an agent for measuring the level of one or more proteins selected from the group consisting of CD110 and CDCP1 or the expression level of a gene encoding the same provides

As used herein, the term "CD110" is an abbreviation of Cluster of Differentiation 110, and is known as a thrombopoietin receptor or myeloproliferative leukemia protein.

The CD110 may include the amino acid sequence represented by SEQ ID NO: 1, and may include an amino acid sequence having 97% or more, specifically 98% or more, more specifically 99% or more homology to the sequence. .

As a sequence consisting of such homology, it is included without limitation as long as it is an amino acid sequence representing a protein that exhibits substantially the same or corresponding efficacy as each protein. In addition, if it is an amino acid sequence consisting of such homology, it is apparent that an amino acid sequence in which some sequences are deleted, modified, substituted or added is also included within the scope of the present invention.

As used herein, the term "CDCP1" is an abbreviation of CUB domain-containing protein 1, which is a large extracellular domain (ECD) including two CUB domains and a smaller intracellular domain (ICD). It may be a transmembrane glycoprotein consisting of a molecular weight of 140 kD. In vivo, CDCP1 is not cleaved under normal physiological circumstances, but cleavage can be induced during tumorigenesis or tissue damage.

The CDCP1 may include the amino acid sequence represented by SEQ ID NO: 2, and may include an amino acid sequence having 97% or more, specifically 98% or more, more specifically 99% or more homology to the sequence. .

In one embodiment of the present invention, the composition for diagnosing cancer metastasis using a blood sample collected from the subject is the level of any one protein selected from the group consisting of ABCA1, IDOL, EpCam, SQLE, and combinations thereof or a gene encoding the same. It may further include an agent for measuring the expression level.

As used herein, the term “ABCA1” is an abbreviation of ATP-binding cassette transporter 1, also called cholesterol efflux regulatory protein (CERP), and is known as a major regulator of cellular cholesterol and phospholipid homeostasis.

The ABCA1 may include the amino acid sequence represented by SEQ ID NO: 3, and may include an amino acid sequence having 97% or more, specifically 98% or more, more specifically 99% or more homology to the sequence. .

As used herein, the term “IDOL” is an abbreviation for Increased Degradation of LDL Receptor Protein, and is known as a ubiquitin ligase that ubiquinates LDL receptors.

The IDOL may include the amino acid sequence represented by SEQ ID NO: 4, and may include an amino acid sequence having 97% or more, specifically 98% or more, more specifically 99% or more homology to the sequence. .

As used herein, the term “EpCAM” is an abbreviation of an epithelial cell adhesion molecule, and is known as a transmembrane glycoprotein that mediates ^{Ca 2+ -independent isoform cell-cell adhesion in the epithelium.}

The EpCAM may include the amino acid sequence represented by SEQ ID NO: 5, and may include an amino acid sequence having 97% or more, specifically 98% or more, more specifically 99% or more homology to the sequence. .

As used herein, the term “SQLE” is an abbreviation for squalene epoxidase, also called squalene monooxygenase. It is known as an enzyme that oxidizes squalene to 2,3-oxidosqualene using oxygen and NADPH. In addition, the enzyme is known to catalyze the oxygenation step of the sterol biosynthesis pathway as one of the rate-limiting enzymes in sterol biosynthesis.

The SQLE may include the amino acid sequence represented by SEQ ID NO: 9, and may include an amino acid sequence having 97% or more, specifically 98% or more, more specifically 99% or more homology to the sequence. .

The gene encoding each protein of the present invention is a nucleotide sequence encoding the amino acid described in each SEQ ID NO, and has 97% or more, specifically 98% or more, more specifically 99% or more homology with the sequence. It may include a nucleotide sequence.

In one embodiment of the present invention, the cancer may be a solid cancer, and the solid cancer may be selected from the group consisting of colorectal cancer, lung cancer, breast cancer, pancreatic cancer, kidney cancer, liver cancer, stomach cancer, uterine cancer and prostate cancer. not limited In addition, specifically, the cancer may be colorectal cancer, and more specifically, it may be colorectal cancer of high cholesterol origin.

As used herein, the term "agent for measuring the level of a protein" may refer to an agent used in a method for measuring the level of a target protein included in a sample.

The agent for measuring the level of the protein may be an antibody that specifically binds to the protein. Specifically, the preparation is western blotting, ELISA (enzyme linked immunosorbent assay), radioimmunoassay (RIA), radioimmunodiffusion, Ouchterlony immune diffusion method, rocket immunoelectrophoresis (rocket immunoelectrophoresis), immunohistochemical staining, immunoprecipitation assay, complement fixation assay, immunofluorescence, immunochromatography, FACS (fluorescenceactivated cell sorter) analysis) and an antibody used in methods such as protein chip technology assay, but is not limited thereto.

The "antibody" refers to a proteinaceous molecule capable of specifically binding to an antigenic site of a protein or peptide molecule. The form of the antibody is not particularly limited, and if it consists of a polyclonal antibody, a monoclonal antibody, or antigen-binding property, a part thereof is also included in the antibody of the present invention, and all immunoglobulin antibodies may be included. In addition, special antibodies such as humanized antibodies may be included. In addition, the antibody includes functional fragments of antibody molecules as well as complete forms having two full-length light chains and two full-length heavy chains. A functional fragment of an antibody molecule means a fragment having at least an antigen-binding function, and may be Fab, F(ab'), F(ab') ₂ and Fv.

As used herein, the term "agent for measuring the expression level of a gene" refers to an agent used in a method for measuring the level of mRNA transcribed from the target gene in order to check whether the target gene contained in the sample is expressed. can mean Specifically, the preparation is PCR, RT-PCR, quantitative real time PCR (quantified real time PCR), competitive RT-PCR (Competitive RT-PCR), real time quantitative RT-PCR (RT-PCR), RNase protection assay (RPA) ; RNase protection assay), Northern blotting, may include a primer or probe capable of specifically binding to a target gene or mRNA thereof used in methods such as DNA chip analysis, but is not particularly limited thereto does not In addition, the gene expression may be mRNA.

The "primer" refers to a short nucleic acid sequence capable of forming a base pair with a template complementary to a target nucleic acid sequence and serving as a starting point for template strand copying. The primer may have a short free 3' hydroxyl group. Primers can initiate DNA synthesis in the presence of reagents for polymerization (ie, DNA polymerase or reverse transcriptase) and four different nucleoside triphosphates in appropriate buffers and temperatures. In addition, the primer may consist of 15 to 35 nucleotides, and may consist of 20 to 25 nucleotides.

The "probe" may be a probe capable of complementary binding to one gene selected from the group consisting of CD110, CDCP1, ABCA1, IDOL, EpCam, SQLE, and combinations thereof, and complementary to each gene As far as possible, the nucleotide sequence of the probe is not limited. The probe may consist of 15 to 35 nucleotides, and may consist of 20 to 25 nucleotides.

Another aspect of the present invention provides a kit for diagnosing cancer metastasis or recurrence using a blood sample collected from a subject, including a composition for diagnosing cancer metastasis or recurrence using a blood sample collected from the subject.

The kit includes a polymerase chain reaction (PCR) kit, a reverse transcription polymerase chain reaction (RT-PCR) kit, a DNA chip kit, an ELISA (Enzyme-linked immunosorbent assay) kit, a protein chip kit, a rapid kit or MRM (Multiple) kit. reaction monitoring) kit. Specifically, the kit for measuring the mRNA expression level of a gene may be a kit including essential elements necessary for performing RT-PCR. The RT-PCR kit includes a test tube or other suitable container, in addition to each primer pair specific for a gene encoding any one protein selected from the group consisting of CD110, CDCP1, ABCA1, IDOL, EpCam, SQLE, and combinations thereof; Reaction buffers (pH and magnesium concentrations vary), deoxynucleotides (dNTPs), enzymes such as Taq-polymerase and reverse transcriptase, DNase, RNAse inhibitors, DEPC-water, sterile water, etc. can In addition, a primer pair specific for a gene used as a quantitative control may be included.

In addition, the kit of the present invention may include essential elements necessary for performing the DNA chip analysis method. The kit for DNA chip analysis may include a substrate to which cDNA corresponding to a gene or fragment thereof is attached as a probe, and reagents, agents, enzymes, etc. for preparing a fluorescently labeled probe. In addition, the substrate may include cDNA corresponding to a quantitative control gene or a fragment thereof.

In addition, the kit of the present invention is a protein chip analysis kit for measuring the level of a protein expressed from a gene encoding any one protein selected from the group consisting of CD110, CDCP1, ABCA1, IDOL, EpCam, SQLE, and combinations thereof. The kit may be, but is not particularly limited thereto, and may include a substrate, an appropriate buffer, a secondary antibody labeled with a chromogenic enzyme or fluorescent material, a chromogenic substrate, and the like for immunological detection of the antibody. Although the substrate is not particularly limited thereto, a nitrocellulose membrane, a 96-well-plate synthesized from polyvinyl resin, a 96-well plate synthesized from polystyrene resin, glass slide glass, etc. may be used, and the chromogenic enzyme is specifically used for this. Although not limited, peroxidase and alkaline phosphatase may be used, and the fluorescent material is not particularly limited thereto, but may be FITC, RITC, etc., and the color substrate solution is not particularly limited thereto, but ABTS ( 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) or OPD (o-phenylenediamine), TMB (tetramethyl benzidine).

In another aspect of the present invention, measuring the level of any one or more proteins selected from the group consisting of CD110 and CDCP1 or the expression level of a gene encoding the same in a blood sample isolated from an individual having cancer; And it provides an information providing method for diagnosing cancer metastasis, comprising determining a cancer metastasis state when the measured protein level or the expression level of a gene encoding the same is higher than that of an individual without cancer metastasis.

In one embodiment of the present invention, the information providing method for diagnosing cancer metastasis is the level of any one protein selected from the group consisting of ABCA1, IDOL, EpCam, SQLE, and combinations thereof in a blood sample isolated from an individual having cancer Or it may further comprise the step of measuring the expression level of the gene encoding it.

In one embodiment of the present invention, in the method for providing information for diagnosing cancer metastasis, the level of any one or more proteins selected from the group consisting of ABCA1 and SQLE measured above or the expression level of a gene encoding the same is not metastasized to cancer. A case that is low compared to the individual is determined as a cancer metastasis state, and the measured level of any one or more proteins selected from the group consisting of IDOL and EpCam or the expression level of a gene encoding the same is compared with an individual without cancer metastasis It may further include the step of determining the high case as a cancer metastasis state.

In addition, according to the present invention, measuring the level of any one or more proteins selected from the group consisting of CD110 and CDCP1 or the expression level of a gene encoding the same in a blood sample isolated from an individual having cancer; And it is possible to provide a method for diagnosing cancer metastasis comprising the step of determining a cancer metastasis state when the measured protein level or the expression level of a gene encoding the same is higher than that of an individual without cancer metastasis.

As used herein, the term “cancer metastasis” means that tumor cells of cancer migrate from an original site of occurrence, settle in another place, and proliferate. In the present invention, the cancer may be a solid cancer, and the solid cancer may be selected from the group consisting of colon cancer, lung cancer, breast cancer, pancreatic cancer, kidney cancer, liver cancer, stomach cancer, uterine cancer and prostate cancer. Specifically, it may be colorectal cancer, and more specifically, it may be colorectal cancer of high cholesterol origin, but is not limited thereto. Specifically, colon cancer cells, unlike normal cells, grow while invading surrounding tissues, so when they encounter tube-shaped structures such as blood vessels or lymphatic vessels, they can move to other places via blood or lymph. When colon cancer cells that have migrated in this way establish themselves in other organs or tissues, they divide and grow again, which is called metastasis of colorectal cancer.

In another aspect of the present invention, measuring the level of any one or more proteins selected from the group consisting of CD110 and CDCP1 or the expression level of a gene encoding the same in a blood sample isolated from a subject undergoing tumor treatment; And measuring the level of the measured protein or the expression level of the gene encoding the same; And it provides an information providing method for diagnosing cancer recurrence, comprising the step of judging a cancer recurrence state when the measured level of the protein or the expression level of the gene encoding the same is higher than that of an individual without cancer recurrence.

In one embodiment of the present invention, the method for providing information for diagnosing cancer recurrence is one protein selected from the group consisting of ABCA1, IDOL, EpCam, SQLE, and combinations thereof in a blood sample isolated from a subject undergoing tumor treatment. It may further comprise the step of measuring the level or the expression level of the gene encoding the same.

In one embodiment of the present invention, the information providing method for diagnosing cancer recurrence is a method in which the level of any one or more proteins selected from the group consisting of ABCA1 and SQLE or the expression level of a gene encoding the measured level of cancer does not recur. A case that is low compared to an individual is determined as a cancer recurrence state, and the level of any one or more proteins selected from the group consisting of IDOL and EpCam or the expression level of a gene encoding the measured level is compared with an individual in which cancer has not recurred. It may further include the step of determining the high case as a cancer recurrence state.

In addition, according to the present invention, measuring the level of any one or more proteins selected from the group consisting of CD110 and CDCP1 or the expression level of a gene encoding the same in a blood sample isolated from a subject undergoing tumor treatment; And it is possible to provide a method for diagnosing cancer recurrence comprising the step of judging a cancer recurrence state when the measured level of the protein or the expression level of a gene encoding the same is higher than that of an individual without cancer recurrence.

As used herein, the term “cancer recurrence” refers to a case in which a tumor of the same type is expressed again in the same site or occurs in a different organ after tumor treatment. In the present invention, the cancer may be a solid cancer, and the solid cancer may be selected from the group consisting of colon cancer, lung cancer, breast cancer, pancreatic cancer, kidney cancer, liver cancer, stomach cancer, uterine cancer and prostate cancer. Specifically, it may be colorectal cancer, and more specifically, it may be colorectal cancer of high cholesterol origin, but is not limited thereto.

In one embodiment of the present invention, the metastasis or recurrence of colorectal cancer may be caused by cholesterol or by inhibition of SQLE expression.

In one embodiment of the present invention, the inhibition of SQLE expression may be induced by a substance that inhibits the level of protein or gene expression, specifically SQLE short hairpin RNA (shSQLE) and small interfering RNA (siSQLE). ) may be used, but is not limited thereto.

As used herein, the term “diagnosis” refers to ascertaining the presence or characteristics of a pathological condition. For the purposes of the present invention, diagnosis is to determine whether or not cancer has metastasized or has metastasis and whether or not cancer has recurred or whether cancer is likely to recur.

As used herein, the term “individual” may refer to any animal, including humans, that has metastasized or is likely to metastasize on the premise that cancer has occurred. The animal may be a mammal, such as a cow, a horse, a sheep, a pig, a goat, a camel, an antelope, a dog, or a cat, in need of treatment for symptoms similar to those of a human as well as humans, but is not limited thereto.

In one embodiment of the present invention, since the "blood sample" isolated from the subject with cancer is representative of a sample obtained by a non-invasive method, any sample obtainable by a non-invasive method is used without limitation can be Specifically, the blood sample may include circulating tumor cells (CTCs) in the blood.

Another aspect of the present invention is for diagnosing cancer metastasis or recurrence using a blood sample collected from an individual comprising an agent for measuring the level of one or more proteins selected from the group consisting of ABCA1 and IDOL or the expression level of a gene encoding the same A composition is provided.

The composition for diagnosing cancer metastasis or recurrence may further include any one protein selected from the group consisting of EpCam, CD110, CDCP1, SQLE, and combinations thereof or a gene encoding the same.

In this case, ABCA1, IDOL, EpCam, CD110, CDCP1 and SQLE are the same as described above in the composition for diagnosing cancer metastasis or recurrence.

Another aspect of the present invention provides a kit for diagnosing cancer metastasis or recurrence using a blood sample collected from a subject, including a composition for diagnosing cancer metastasis or recurrence using a blood sample collected from the subject.

In another aspect of the present invention, measuring the level of one or more proteins selected from the group consisting of ABCA1 and IDOL or the expression level of a gene encoding the same in a blood sample isolated from an individual having cancer; And when the measured level of ABCA1 protein or the expression level of a gene encoding the same is low compared to a non-cancerous individual, it is determined as a cancer metastasis state, and the measured level of IDOL protein or expression of a gene encoding the same It provides a method of providing information for diagnosing cancer metastasis, comprising determining a cancer metastasis state when the level is higher than that of an individual who has not metastasized.

In one embodiment of the present invention, the information providing method for diagnosing cancer metastasis is the level of any one protein selected from the group consisting of EpCam, CD110, CDCP1, SQLE, and combinations thereof in a blood sample isolated from an individual having cancer Or measuring the expression level of the gene encoding it; And a case in which the measured level of SQLE protein or the expression level of a gene encoding the same is lower than that of an individual without cancer metastasis is determined as a cancer metastasis state, and the measured EpCam, CD110, CDCP1, and a combination thereof. The method may further include determining a cancer metastasis state when the level of any one protein selected from or the expression level of a gene encoding the same is higher than that of an individual without cancer metastasis.

In addition, according to the present invention, measuring the level of one or more proteins selected from the group consisting of ABCA1 and IDOL or the expression level of a gene encoding the same in a blood sample isolated from an individual having cancer; And when the measured level of ABCA1 protein or the expression level of a gene encoding the same is low compared to a non-cancerous individual, it is determined as a cancer metastasis state, and the measured level of IDOL protein or expression of a gene encoding the same A method for diagnosing cancer metastasis can be provided, comprising determining a cancer metastasis state when the level is high compared to an individual who has not metastasized.

In another aspect of the present invention, measuring the level of one or more proteins selected from the group consisting of ABCA1 and IDOL or the expression level of a gene encoding the same in a blood sample isolated from a subject undergoing tumor treatment; and a case where the measured level of ABCA1 protein or the expression level of a gene encoding the same is low compared to a non-cancerous individual as a cancer recurrence state, and the measured level of IDOL protein or expression of a gene encoding the same It provides a method for providing information for diagnosing cancer recurrence, comprising determining a cancer recurrence state when the level is higher than that of an individual with no recurrence of cancer.

In one embodiment of the present invention, the method for providing information for diagnosing cancer recurrence includes EpCam, CD110, CDCP1, SQLE, and any one protein selected from the group consisting of a combination thereof in a blood sample isolated from a subject undergoing tumor treatment. measuring the level or the expression level of a gene encoding the same; and a case in which the measured level of SQLE protein or the expression level of a gene encoding the same is low compared to an individual in which cancer does not recur is determined as a cancer recurrence state, and the measured EpCam, CD110, CDCP1, and a combination thereof. It may further include the step of judging a cancer recurrence state when the level of any one protein selected from or the expression level of a gene encoding the same is high compared to an individual in which cancer does not recur.

In addition, according to the present invention, measuring the level of one or more proteins selected from the group consisting of ABCA1 and IDOL or the expression level of a gene encoding the same in a blood sample isolated from a subject undergoing tumor treatment; and a case where the measured level of ABCA1 protein or the expression level of a gene encoding the same is low compared to a non-cancerous individual as a cancer recurrence state, and the measured level of IDOL protein or expression of a gene encoding the same It is possible to provide a method for diagnosing cancer recurrence, comprising judging a cancer recurrence state when the level is higher than that of a non-cancerous individual.

As used herein, the term "sample" is isolated from a patient with cancer and is selected from the group consisting of CD110, CDCP1, ABCA1, IDOL, EpCam, and combinations thereof to measure the expression level of a gene encoding a protein It means a direct target, and specifically, the sample may be blood, cancer cells, or cancer tissue, and specifically may be circulating tumor cells in the blood, but is not limited thereto.

As used herein, the term "circulating tumor cell" is a tumor cell found in the peripheral blood of a malignant tumor patient, and it is known that it can originate from both a primary tumor and a tissue in which metastasis has occurred.

In one embodiment of the present invention, treatment with cholesterol in colorectal cancer cell lines (HCT116 and HT29) increases the expression of cancer stem cell markers (EpCAM) and cancer cell markers (CD110 and CDCP1) like circulating tumor cells, and decreases the expression of SQLE It was confirmed that (see FIGS. 3a and 3b).

In another embodiment of the present invention, the expression of cancer stem cell markers (EpCAM) and cancer cell markers (CD110 and CDCP1) is increased in a blood sample of a high cholesterol diet cancer metastasis animal model and/or a cancer metastasis animal model in which the expression of SQLE is suppressed. And it was confirmed that the expression of SQLE is reduced (see FIGS. 6a to 6d).

These experimental results indicate that circulating tumor cells having SQLE, EpCam, CD110 and CDCP1 markers are present in the blood of an animal model of cancer metastasis, which is at least one selected from the group consisting of SQLE, EpCAM, CD110 and CDCP1. This suggests that a protein or a gene encoding the same can be utilized for diagnosing cancer metastasis.

Another aspect of the present invention provides a composition using a blood sample collected from an individual comprising an agent for measuring the level of one or more proteins selected from the group consisting of CD110 and CDCP1 or the expression level of a gene encoding the same. Provided is a use for diagnosing relapse.

Another aspect of the present invention provides a composition using a blood sample collected from an individual comprising an agent for measuring the level of one or more proteins selected from the group consisting of CD110 and CDCP1 or the expression level of a gene encoding the same. Provided is a use for manufacturing a medicament for diagnosis of recurrence.

The composition using a blood sample collected from an individual including a preparation for measuring the level of one or more proteins selected from the group consisting of CD110 and CDCP1 or the expression level of a gene encoding the same is as described in the composition for diagnosing cancer metastasis or recurrence .

Another aspect of the present invention provides the use of a composition comprising an agent for measuring the level of one or more proteins selected from the group consisting of ABCA1 and IDOL or the expression level of a gene encoding the same for diagnosing cancer metastasis or recurrence do.

Another aspect of the present invention is the use of a composition comprising an agent for measuring the level of one or more proteins selected from the group consisting of ABCA1 and IDOL or the expression level of a gene encoding the same for preparing a medicament for diagnosing cancer metastasis or recurrence provides

A composition comprising an agent for measuring the level of one or more proteins selected from the group consisting of ABCA1 and IDOL or the expression level of a gene encoding the same is as described above in the composition for diagnosing cancer metastasis or recurrence.

Hereinafter, the present invention will be described in more detail through examples. These Examples are for explaining the present invention in more detail, and the scope of the present invention is not limited to these Examples.

Experimental Example 1. Confirmation of increase in survival of colorectal cancer cell lines by cholesterol treatment or inhibition of squalene epoxidase (SQLE) expression

Experimental Example 1.1. Preparation of SQLE expression inhibitory substances

In order to investigate the intracellular function by suppressing the expression of the SQLE gene, siRNA (siSQLE) consisting of 25 nucleotides in the nucleotide sequence of the SQLE gene was prepared. siCont was prepared as siRNA for control experiments.

The nucleotide sequence and amino acid sequence of SQLE are shown in SEQ ID NO: 6 and SEQ ID NO: 9, respectively, and the nucleotide sequences of siCont and siSQLE are shown in SEQ ID NO: 7 and SEQ ID NO: 8, respectively.

Experimental Example 1.2. cell culture

To check the viability of colorectal cancer cell lines by treatment with cholesterol or inhibition of SQLE expression, colon cancer cell lines HCT116 and HT29 cell lines were cultured. Each of the cell lines was treated with a medium containing 10% fetal bovine serum, 100 mg/ml streptomycin, and 100 IU/ml ampicillin in ultra-low attachment plates (Corning, USA) for 72 hours. Cell line samples were treated with cholesterol and siRNA at 20 μM daily during the incubation period.

Experimental Example 1.3. Cell viability measurement

The cultured cell line sample was observed with an optical microscope at a magnification of 40 times, and the photomicrograph is shown in FIG. 1 .

In addition, in order to statistically confirm the increase in survival of colorectal cancer cell lines by cholesterol treatment or SQLE expression inhibition, the following method was performed based on the MTT method. The cell line cultured in Experimental Example 1.2. was transferred to a 96-well-plate, treated with 2 mg/ml MTT, reacted at 37° C. for 2 hours, and then the reaction was stopped with DMSO. After reading the absorbance with a microplate reader (570 nm), the cell death/survival was measured by converting it into a percentage. In addition, apoptosis/survival of colorectal cancer cells was confirmed in the same manner for the cholesterol-untreated group and 20 μM siCont-treated group as a control group.

As a result, in the colorectal cancer cell line treated with cholesterol and siSQLE, the cell viability was significantly increased compared to the control group (FIG. 1).

Referring to the photomicrograph of FIG. 1 , it was observed that cell aggregate formation indicating cell survival was increased in colorectal cancer cell lines cultured on non-adherent plates, which were cholesterol and siSQLE-treated colorectal cancer cell lines treated with anoikis (anoikis). means it didn't work out well.

Anoikis is a form of apoptosis and refers to cell death that occurs when cells are not attached to the cell substrate or are attached to a place other than their original location. Cells that have acquired resistance to Anoikis can survive in suspension Alternatively, growth may become possible at a site other than the original site, and as a result, metastasis of cancer may be induced.

Experimental Example 2. Confirmation of expression of cancer cell markers and cholesterol inflow or release related proteins in colorectal cancer cell lines according to cholesterol treatment

Colorectal cancer cell lines (HCT116 and HT29) were cultured for 72 hours in non-adherent plates with 20 μM cholesterol daily treatment. After the culture was terminated, the gene expression levels of cancer cell markers (CD110 and CDCP1), cholesterol influx related protein (IDOL) and cholesterol release related protein (ABCA1) were confirmed using quantitative polymerase chain reaction (Quantitative PCR) analysis, The results are shown in FIG. 2 .

Referring to FIG. 2 , in the cholesterol-treated HCT116 colon cancer cell line, the expression levels of SQLE and ABCA1 were significantly decreased, and the expression levels of CD110 and IDOL were significantly increased compared to the cholesterol-untreated control group. In addition, in the cholesterol-treated HT29 colorectal cancer cell line, the expression levels of SQLE and ABCA1 were significantly decreased, and the expression levels of CDCP1 and IDOL were significantly increased compared to the cholesterol-untreated control group.

Experimental Example 3. Confirmation of expression levels of cancer stem cell markers, cancer cell markers and SQLE in colorectal cancer cell lines according to cholesterol treatment

Colorectal cancer cell lines (HCT116 or HT29) were cultured for 72 hours with daily treatment of 20 μM cholesterol on non-adherent plates, and then washed 3 times with FACS staining buffer (0.05% BSA in PBS buffer). Cells were collected, fixed in BD Cytofix Fixation solution for 15 minutes, and washed once with FACS staining buffer. The washed cell pellet was diluted in BD Permeabilization buffer and reacted for 5 minutes. After washing with FACS staining buffer for 3 times, the control (CD45), cancer stem cell marker (EpCAM), cancer cell marker (CD110 and CDCP1) and SQLE antibodies were treated and reacted for 60 minutes. After the reaction, the FACS staining buffer After washing with a flow cytometer (BD Biosciences), the expression levels of EpCam, CD110, CDCP1 and SQLE were analyzed, and the results in colorectal cancer cell lines (HCT116 and HT29) are shown in FIGS. 3a and 3b, respectively. Antibodies used are shown in Table 1.

[Table 1]

As a result, referring to FIGS. 3A and 3B , the expression of EpCAM, CD110 and CDCP1 was increased, and the expression of SQLE was decreased in the cholesterol-treated colorectal cancer cell lines (HCT116 and HT29).

Experimental Example 4. in vivo Confirmation of cancer metastasis according to cholesterol treatment and SQLE expression level in mouse animal model using luciferase method

Experimental Example 4.1. Preparation of cancer metastasis animal model

A schematic diagram of a method for preparing an animal model of cancer metastasis is shown in FIG. 4A . Specifically, mice (SCID, Immune-deficient mice, 5 weeks old) were fed a 2% high cholesterol diet every day for 30 days to keep the cholesterol levels in the mouse serum high. Control mice were bred by providing the same general diet.

Meanwhile, a cancer cell line having a normal level of SQLE gene (siCont) and a cancer cell line having a knock-out SQLE gene (siSQLE) were prepared. The SQLE gene knock-out (knock-out) cancer cell line was obtained by infecting HT29 or HCT116 cell lines with SQLE Mission shRNA lentivirus transducing particles (Sigma: TRCN0000046155, TRCN0000046157). 100 μl of the cancer cell line (5 x 10 ⁶ cells/ml) was subcutaneously injected dorsally into a mouse (SCID, 5 weeks old), and when the tumor size reached a maximum diameter of 0.5 x 10 ² mm ³ , the tumor was excised. On the other hand, after tumor resection, the mice were further bred for 120 to 130 days, and the condition was monitored.

The resected tumor was digested into a single cell suspension, and then the cell suspension was continuously filtered using sterile gauze and a 70 μm cell strainer (Corning, 352350). Thereafter, CDCP1 and CD110-rich cells were sorted through flow cytometry and the cell density ^{was concentrated to 5 X 10 5} cells/ml.

25 μl of the concentrated CDCP1 and CD110-rich cell sample (5 X 10 ⁵ cells/ml) was suspended in PBS (phosphate-buffered saline) containing 1% FBS (fetal bovine serum; Corning, 35-015-CV). Thus, cancer metastasis animal model mice were prepared by injecting the mice into the colon through laparotomy in the mice bred for 30 days. For spontaneous (orthotopic) metastasis assay, mice were further bred for 120 to 130 days and their status was monitored.

The cancer metastasis animal model mice were divided into (i) cholesterol untreated / siSQLE group, (ii) 2% cholesterol / siCont group and (iii) 2% cholesterol / siSQLE group to constitute an experimental group, (iv) cholesterol untreated / siCont group was used as a control group.

All animal experiments were approved by the KRIBB Bioethics Committee.

Experimental Example 4.2. Analysis of serum cholesterol in an animal model of cancer metastasis

To the cancer metastasis animal model mice prepared in Experimental Example 4.1., a 2% high cholesterol diet or a normal diet was provided to measure serum cholesterol while breeding (FIG. 4b). Specifically, in FIG. 4B, (1) is the measurement of serum cholesterol of mice treated with the normal diet for 5 weeks, and (2) is the laparotomy of Experimental Example 4.1. Cholesterol in serum was measured immediately before surgery (about 9 weeks), and in (3), serum cholesterol was measured when the study was completed (about 25 to 26 weeks) in mice fed a high-cholesterol diet or a regular diet. did it

As a result, it was confirmed that the serum cholesterol level was high after 9 weeks in mice fed a 2% cholesterol diet.

Experimental Example 4.3. Confirmation of cancer metastasis in cancer metastasis animal model

At 30 days, 60 days, 90 days, and 120 days after the injection of cancer cells into the cancer metastasis animal model mouse prepared in Experimental Example 4.1., the movement of cancer cells was monitored using ivis (In Vivo Imaging Systems; Perkin elemer). was confirmed (FIGS. 4c to 4f).

As a result, it was confirmed that the degree of cancer metastasis in mice fed a 2% cholesterol diet was significantly increased compared to the control group. In addition, it was confirmed that the degree of cancer metastasis significantly increased even in mice in which SQLE expression was suppressed while providing a general diet.

Experimental Example 5. in vivo Check the level of cancer cell marker (CDCP1) in lung tissue according to the level of inhibition of cholesterol treatment and SQLE expression in

Lung tissue was collected from the cancer metastasis mouse model (5 mice in each experimental group and control group) in which the cancer metastasis of Experimental Example 4.3. The degree of cancer metastasis to lung tissue was measured by performing quantitative calculations using the Image J program. Nuclei were stained with DAPI antibody (blue). The results for the cancer metastasis mouse model transplanted with HT29 are shown in FIG. 5A, and the results for the cancer metastasis animal model transplanted with HCT116 are shown in FIG. 5B. The antibodies used are shown in Table 2.

[Table 2]

Referring to FIGS. 5A and 5B , it was confirmed that the expression of CDCP1 in the lung tissue was significantly increased in the experimental group compared to the control group. Since the CDCP1 is a marker for colon-derived cells, it means that the lung tissue in which the expression of CDCP1 is remarkably increased is metastasized from colorectal cancer.

Experimental Example 6. Confirmation of the level of cancer stem cell marker and cancer cell marker in the blood of cancer metastasis animal model

The levels of cancer stem cell markers (EpCam) and cancer cell markers (CD110 and CDCP1) according to the degree of cholesterol treatment and SQLE expression inhibition were confirmed in the blood samples of the cancer metastasis animal model prepared in Experimental Example 4.1.

Specifically, the experimental group and the control group of the cancer metastasis animal model were bred by providing a 2% high cholesterol diet and a general diet, respectively. Blood was collected from an animal model of cancer metastasis, and the blood sample was treated with a blood cell marker (CD45), a cancer stem cell marker (EpCAM), a cancer cell marker (CD110 and CDCP1), and an antibody specific for SQLE. Then, the expression levels of EpCam, CD110, CDCP1 and SQLE were analyzed using a flow cytometer ( FIGS. 6a and 6c ), and numerically shown in FIGS. 6b and 6d (*, p<0.05; **, p <0.01; ***, p<0.001). The antibodies used are shown in Table 1 above.

As a result, referring to FIGS. 6A to 6D , the expression levels of EpCam, CD110 and CDCP1 were increased compared to the control group in both the HT29-transplanted cancer metastasis animal model experimental group and the HCT116-transplanted cancer metastasis animal model experimental group, and the expression of SQLE level decreased.

These experimental results indicate that circulating tumor cells having EpCam, CD110, and CDCP1 markers exist in the blood of an animal model of cancer metastasis.

Claims (23)

A composition for diagnosing cancer metastasis or recurrence using a blood sample collected from an individual, comprising an agent for measuring the level of one or more proteins selected from the group consisting of CD110 and CDCP1 or the expression level of a gene encoding the same. According to claim 1, Cancer metastasis using a blood sample collected from an individual, further comprising an agent for measuring the level of any one protein selected from the group consisting of ABCA1, IDOL, EpCam, SQLE, and combinations thereof or the expression level of a gene encoding the same or a composition for diagnosing recurrence. According to claim 1, Wherein the cancer is a solid cancer, a composition for diagnosing cancer metastasis or recurrence using a blood sample collected from an individual. 4. The method of claim 3, Wherein the solid cancer is selected from the group consisting of colon cancer, lung cancer, breast cancer, pancreatic cancer, kidney cancer, liver cancer, stomach cancer, uterine cancer and prostate cancer, a composition for diagnosing cancer metastasis or recurrence using a blood sample collected from an individual. According to claim 1, The agent for measuring the level of the protein is an antibody that specifically binds to the protein, a composition for diagnosing cancer metastasis or recurrence using a blood sample collected from an individual. According to claim 1, The agent for measuring the expression level of the gene is a primer or probe that specifically binds to the gene or its mRNA, a composition for diagnosing cancer metastasis or recurrence using a blood sample collected from an individual. According to claim 1, The blood sample comprises a circulating tumor cell in the blood, a composition for diagnosing cancer metastasis or recurrence using a blood sample collected from an individual. A kit for diagnosing cancer metastasis or recurrence using a blood sample collected from an individual, comprising the composition of any one of claims 1 to 7. Measuring the level of any one or more proteins selected from the group consisting of CD110 and CDCP1 or the expression level of a gene encoding the same in a blood sample isolated from an individual having cancer; and A method for providing information for diagnosing cancer metastasis, comprising determining a cancer metastasis state when the measured level of the protein or the expression level of a gene encoding the same is higher than that of a non-cancerous individual. 10. The method of claim 9, Further comprising the step of measuring the level of any one protein selected from the group consisting of ABCA1, IDOL, EpCam, SQLE, and a combination thereof or the expression level of a gene encoding the same in a blood sample isolated from an individual having cancer, A method of providing information for diagnosing cancer metastasis. 11. The method of claim 10, A case in which the level of any one or more proteins selected from the group consisting of ABCA1 and SQLE or the expression level of a gene encoding the same is low compared to an individual without cancer metastasis is determined as a cancer metastasis state, and the measured IDOL And EpCam, when the level of any one or more proteins selected from the group consisting of or the expression level of a gene encoding the same is higher than that of an individual who has not metastasized, it is determined as a cancer metastasis state, information providing method for diagnosing cancer metastasis . 10. The method of claim 9, The method of providing information for diagnosing cancer metastasis, wherein the blood sample includes circulating tumor cells in the blood. Measuring the level of any one or more proteins selected from the group consisting of CD110 and CDCP1 or the expression level of a gene encoding the same in a blood sample isolated from a subject undergoing tumor treatment; and A method for providing information for diagnosing cancer recurrence, comprising determining a cancer recurrence state when the measured protein level or the expression level of a gene encoding the same is higher than that of an individual without cancer recurrence. 14. The method of claim 13, Further comprising the step of measuring the level of any one protein selected from the group consisting of ABCA1, IDOL, EpCam, SQLE, and a combination thereof or the expression level of a gene encoding the same in a blood sample isolated from a subject undergoing tumor treatment , an informational method for diagnosing cancer recurrence. 15. The method of claim 14, A case in which the level of any one or more proteins selected from the group consisting of ABCA1 and SQLE or the expression level of a gene encoding the same is low compared to a non-cancerous individual is determined as a cancer recurrence state, and the measured IDOL And EpCam, when the level of any one or more proteins selected from the group consisting of or the expression level of a gene encoding the same is high compared to a non-cancerous individual as a cancer recurrence state, a method for diagnosing cancer recurrence . 14. The method of claim 13, The method of providing information for cancer recurrence diagnosis, wherein the blood sample includes circulating tumor cells in the blood. A composition for diagnosing cancer metastasis or recurrence using a blood sample collected from an individual, comprising an agent for measuring the level of one or more proteins selected from the group consisting of ABCA1 and IDOL or the expression level of a gene encoding the same. 18. The method of claim 17, EpCam, CD110, CDCP1, SQLE, and cancer metastasis using a blood sample collected from an individual, further comprising an agent for measuring the level of any one protein selected from the group consisting of or the expression level of a gene encoding the same or a composition for diagnosing recurrence. A kit for diagnosing cancer metastasis or recurrence using a blood sample collected from an individual, comprising the composition of claim 17 or 18. Measuring the level of one or more proteins selected from the group consisting of ABCA1 and IDOL or the expression level of a gene encoding the same in a blood sample isolated from an individual having cancer; and A case in which the measured level of ABCA1 protein or the expression level of a gene encoding the same is lower than that of a non-cancerous individual is judged as a cancer metastasis state, and the measured level of IDOL protein or the expression level of a gene encoding the same A method for providing information for diagnosing cancer metastasis, comprising the step of judging a cancer metastasis state when the cancer is higher than that of a non-metastatic individual. 21. The method of claim 20, Measuring the level of any one protein selected from the group consisting of EpCam, CD110, CDCP1, SQLE, and a combination thereof or the expression level of a gene encoding the same in a blood sample isolated from an individual having cancer; and A case in which the measured level of SQLE protein or the expression level of a gene encoding the same is low compared to a non-cancerous individual is determined as a cancer metastasis state, and in the group consisting of the measured EpCam, CD110, CDCP1, and combinations thereof Method for providing information for diagnosing cancer metastasis, further comprising determining a cancer metastasis state when the level of any one selected protein or the expression level of a gene encoding the same is higher than that of a non-cancerous individual . Measuring the level of one or more proteins selected from the group consisting of ABCA1 and IDOL or the expression level of a gene encoding the same in a blood sample isolated from a subject who has undergone tumor treatment; and A case in which the measured level of ABCA1 protein or the expression level of a gene encoding the same is lower than that of a non-cancerous individual is determined as a cancer recurrence state, and the measured level of IDOL protein or the expression level of a gene encoding the same A method for providing information for diagnosing cancer recurrence, comprising the step of judging a cancer recurrence state when the cancer is higher than that of a non-recurring individual. 23. The method of claim 22, Measuring the level of any one protein selected from the group consisting of EpCam, CD110, CDCP1, SQLE, and a combination thereof or the expression level of a gene encoding the same in a blood sample isolated from a subject undergoing tumor treatment; and A case in which the measured level of SQLE protein or the expression level of a gene encoding the same is low compared to a non-cancerous individual is determined as a cancer recurrence state, and in the group consisting of the measured EpCam, CD110, CDCP1, and combinations thereof Method for providing information for diagnosing cancer recurrence, further comprising the step of judging a cancer recurrence state when the level of any one selected protein or the expression level of a gene encoding the same is higher than that of a non-cancerous individual .

Images