CN116162706A - Exosome lncRNA marker group for esophageal squamous carcinoma diagnosis and application thereof - Google Patents

Abstract

The invention discloses an exosome lncRNA marker group for esophageal squamous carcinoma diagnosis and application thereof. The exosome lncRNA marker group consists of lncRNA AC098818.2, RASSF8-AS1 and LINC00958. The exosome lncRNA marker combination can be used for preparing an esophageal squamous carcinoma diagnosis product, is used for noninvasive screening of esophageal squamous carcinoma, improves the early detection rate of esophageal squamous carcinoma patients, performs early intervention, and improves the overall survival rate of esophageal squamous carcinoma patients.

Description

An exosomal lncRNA marker set for the diagnosis of esophageal squamous cell carcinoma and its application

technical field

The invention relates to the technical field of tumor markers, in particular to an exosomal lncRNA marker group for diagnosis of esophageal squamous cell carcinoma and its application.

Background technique

Esophageal squamous cell carcinoma is the sixth most malignant tumor in my country, and its prognosis is poor, with a five-year overall survival rate of less than 20%. Esophageal squamous cell carcinoma is the main pathological type in my country. Due to the lack of symptoms in the early stage, most patients with esophageal squamous cell carcinoma miss the best time for diagnosis and treatment when they are diagnosed. The diagnosis of esophageal squamous cell carcinoma mainly depends on gastroscopic biopsy. Gastroscopy is an invasive operation with low comfort and high cost. For most Chinese people, gastroscopy is not used as a daily physical examination item. Therefore, It is also easy to cause the early lesions of esophageal squamous cell carcinoma to be missed.

Liquid biopsy of humoral tumor markers provides a new tool for solid tumor diagnosis. Traditional tumor markers include squamous cell antigen SCC, cytokeratin 19 fragment CYFRA21-1, carcinoembryonic antigen CEA, etc. However, these tumor markers face problems such as poor specificity and sensitivity when applied to body fluid biopsy, which limits their application. In recent years, the development of new tumor markers, such as blood cell-free DNA (cfDNA), blood circulating miRNA, etc., has been developed in full swing. Long non-coding RNA has been a research hotspot in recent years. It has similar structural characteristics to mRNA, but has no protein coding function. Its role in tumorigenesis and metastasis has been increasingly valued. Studies have shown that long non-coding RNA is expected to be used as a new tumor marker for the diagnosis and prognosis of tumors. However, in the absence of protection, RNA molecules in blood are easily digested by RNase, resulting in instability and are not suitable for liquid biopsy. The discovery of exosomes provides a possibility for blood RNA-based diagnosis.

Exosomes are a type of cell vesicles, which are formed by the invagination of the endoplasmic reticulum membrane and secreted outside the cell. During the formation of exosomes, nucleic acid molecules and proteins in cells can be packaged and transferred between cells. Due to the protective effect of exosome surface proteins, the exosome contents have better stability. Therefore, lncRNA in exosomes is expected to become a new marker for liquid biopsy. Previous studies have shown that exosomal lncRNA can be used for non-invasive diagnosis of malignant tumors such as non-small cell lung cancer and gastric cancer, but there are still few reports on the use of exosomal lncRNA in the diagnosis of esophageal squamous cell carcinoma.

Contents of the invention

In order to solve the above technical problems, the present invention provides an exosomal lncRNA marker group for the diagnosis of esophageal squamous cell carcinoma and its application.

The present invention is realized by adopting the following technical solutions.

An exosomal lncRNA marker group for the diagnosis of esophageal squamous cell carcinoma, the exosomal lncRNA marker group consists of lncRNAAC098818.2, RASSF8-AS1, LINC00958.

A diagnostic model based on the above exosome lncRNA marker group, the diagnostic model is EXPAC098818.2*0.74429+EXP LINC00958*1.24591+EXP RASSF8-AS1*1.27292.

An application of the above-mentioned exosome lncRNA marker group in the preparation of products for diagnosing esophageal squamous cell carcinoma.

The application has the following beneficial effects.

The present invention establishes a combined diagnostic model of esophageal squamous cell carcinoma based on three lncRNA molecules, which has high diagnostic value and better diagnostic performance than traditional blood tumor markers, and provides a new index for noninvasive screening of esophageal squamous cell carcinoma.

Description of drawings

Figure 1 is a diagram of the quantitative detection results of five preliminary candidate lncRNAs in serum, tissue and cell lines;

Figure 2 is a diagram of the results of exosome isolation and identification of normal people and patients with esophageal squamous cell carcinoma of the present invention (Among them, A: Exosome morphological characteristics under the electron microscope; B: Exosome particle size-concentration curve);

Figure 3 is a diagram of the stability detection results of exosome lncRNA of the present invention (Among them, A: the stability detection result diagram of exosome lncRNA after being treated at different environmental temperatures; B: the stability detection result of exosome lncRNA after RNase treatment Figure; C: Exosomal lncRNA stability test results after repeated freezing and thawing; D: Exosomal lncRNA stability test results after acid/alkali treatment);

Figure 4 is a comparison chart of the expression level of exosome lncRNA before and after operation of the present invention;

Fig. 5 is the result graph of the training set stage of the present invention (among them, A: a comparison chart of exosomal lncRNA expression levels between patients with esophageal squamous cell carcinoma and healthy people; B: ROC curve graph of the Index I diagnostic model; C: Index II diagnostic model ROC curve graph);

Fig. 6 is the result graph of the verification set stage of the present invention (Among them, A: comparison chart of exosomal lncRNA expression levels between patients with esophageal squamous cell carcinoma and healthy people; B: ROC curve graph of Index I diagnostic model; C: Index II diagnostic model ROC curve graph).

Detailed ways

Below in conjunction with embodiment this patent application is further described.

1. The present invention screened the serum exosomal lncRNA differentially expressed between patients with esophageal squamous cell carcinoma and the healthy control group. After strict screening criteria and large sample verification, a molecular set composed of three lncRNAs was finally obtained, namely lncRNAAC098818. 2. RASSF8-AS1, LINC00958.

The screening process is as follows: 1. Collect serum from patients with esophageal squamous cell carcinoma and healthy controls, 4 cases in each group, 2ml of serum in each case. Serum exosomes were extracted, the shape of exosomes was identified by transmission electron microscopy, and exosome RNA was extracted with an exosome total RNA extraction kit, and the quality of RNA was detected by a nanodrop spectrophotometer, and the LncRNA_8×60k expression profile chip of Arraystar Company was used The expression profile of lncRNA was quantitatively detected, and the abnormally expressed lncRNA in the serum of patients with esophageal squamous cell carcinoma was screened by unpaired T test (FDR<0.005, Foldchange>2.0). 2. In addition, 5 pairs of esophageal squamous cell carcinoma and normal esophageal epithelial tissues were taken respectively, and the total RNA in the tissues was extracted with Trizol reagent after grinding, and the lncRNA in the esophageal squamous cell carcinoma tissue was screened in the same way, and the abnormally expressed lncRNA was detected by the paired T test method. Screening (FDR<0.005, Foldchange>2.0); 3. Take the intersection of the abnormally expressed lncRNA in the serum and tissue of patients with esophageal squamous cell carcinoma, and use FDR<0.005, Foldchange>5.0 as the screening conditions, and obtain 5 lncRNAs in patients under this condition The up-regulated lncRNAs AC098818.2, RASSF8-AS1, LINC00958, GMDS-DT, AL591721.1 in both serum and cancer tissues were identified as preliminary candidate lncRNAs; 4. Further serum samples (n= 20/group, Figure 1A), esophageal squamous cell carcinoma tissue and paired normal esophageal epithelial tissue (n=30, Figure 1B), as well as esophageal squamous cell carcinoma cell lines (TE1, TE13, KYSE150, KYSE140) and normal esophageal epithelial cell line Het -1A was validated by qRT-PCR (Figure 1C). The results showed that only AC098818.2, RASSF8-AS1, and LINC00958 were overexpressed in the serum samples of esophageal squamous cell carcinoma patients, esophageal squamous cell carcinoma tissues and esophageal squamous cell carcinoma cell lines, so they were identified as the final candidate lncRNAs. See below for relevant RNA extraction methods and qRT-PCR quantitative detection methods.

Candidate lncRNA sequence information is as follows:

ENST00000564925.1 (AC098818.2) (SEQ ID NO.1)

gacggttccggctcttgtcacgtggagggggaccaagggaggtgattgtg

ggggttggggactagggcaaagtcgggggtcccgggtccggggttgcgcc

gggagggtactggccaggcccgacttcaagcagggaggtggcccgggact

ggctcccgcgctggtcacgcaccggggtccgtcatctcgaccgggagtcg

ctgtgccactctaggggcttaaaaatgccgccaacaactttacccctgct

gaggaacttctcgcttcgtttcttagccagttttccaggccttctccctt

gagagtctccttccgctcgcccaccctgagcgtagggaggtgggagcagga

cggaactggagtggattccagtaactggacttctaaaccggctcggtgtc

gtcgcgctctgtagcggacgcttgtgctgggaagcgttttgcagtttata

aattacttaggagtactttaaacatctcacgcctgctgtaccacagccat

gtgggggtattcagaacaggtattaaggtcgttttgcagaccattcatca

ctcttggctaaaagaacttaacggagtcgcccaaggtcacacagctggct

cgtggtaattggggaccattctttgattcataattttgtgcttttggcca

ccctgaaaaagtaaggtcagatcgttgaaaaacgggcagatttaaggctg

tttggaagcaaaagcgaccactgcgttcccctcaaatcaaagagatttgc

accccctaccaaaaaaaaaaaaaaaaaaaaaagccctgttgtatcaattt

cataaactgcaggagtctcccgactgcagggtgaagcaacaaaataaata

tgggtgtaaaacattcaggagtggtagggaatggcaaactgtagagcctg

tgtaaagagagcagccaattctcagctccagcctacttttccacaaaaga

agataggcacacacactgtgcccagaggttccagtttcttaagagaacca

aaaacctggaactgttttctaatgtggcatttccctaatttttaaaatat

agatcaaataaattgtatctgcaggactccagtatgcaccttctatgaaa

gtagatgaatgtggttctatccccaccccctatttctttattctttttaat

gagataacgggtagaaacattagaaaatgtgcttagacaaaaactttgaa

cttaaaatccttgttctttcttccctgctatttgaagagaagctatttct

gccaacactaaaatagtgctctggatttaggcattctgctacataaccc

actaaactaaaattcttcatccagagacctgacgtttaaagtagattcca

gcttagtggaagtgagattatatgtcagatgattgatgtggtgtggcagt

tggggatttccattattagaaacaagctgaccactttcgttgctgtggggct

tcattgtggagtctcattcctaaatttgaattgttttttaagcagaataa

tggaaagattaagtcatttcatgcaattcaaaactggagtcagacaaaca

tacaaagtttgggcctgatataacttcacctgatcacaacaagcctgcta

taataagtttcaaagttagtttacgaggatcacttgaggccaggaggttca

aaaccagcatggtcaacataacaagacctcatctctttaaaaaaaaaaaaa

aaaaaaaattaaaaattagccaggtatggtggtggtgcacctgtaatccc

agctactcaggaggctgaggcaggaggatcgcttgaaccaaggagttcaa

ggctgcagtgcgctatgatggcactggtgcactccaggctgggtgtctct

taaaaacaaacaaaagtttgaccagtcaaagtggcttgcacctataatcc

cagcactttgggaagccgaggtgggtggattgcttgaactcaggagttca

agaccaacctgggcaatatggtgagaccttccatctctatcaaaaataca

aaaattagctgggcttggtggggtggcgcaggcctgtggtccctgctact

tgggaagctgaggtgcgtggatcactggagccaaggaagtcgaggctgca

gtgagccatgatagcatcagctgcactccagcctggacaacagaggaaga

cactgtctctaagtaaataaataagtcaatttacaattaatgtggtaaag

attacktgtatgtaccaatgtaacttgacttttgatattgaaatgtataa

ttataataaatgtataattaaatgtgtaataatgtatagaccccctctga

aagacgagggtaacaccaagENST00000545729.6 (RASSF8-AS1) length = 3188 (SEQ ID NO. 2) aaactcccttgccaaggacactggggaggaagcagaaaagcaaagaccaagctcgtacttttcagtccgcggcttgtgtgccgcgaggagaggcgcaaaggggtgacacaccag gaaagcaggtagcaggcaagagtgtggttatcgttggaccaggattcaggatctgactgagtagctgactcttaatccagtttgtgttgatcaccaggaggatctggacctcagtatttaccacatctagagctataatcacatccctgttaagccttcctgagatcacaccaaatgtccttgatgggaattgtaaacaacccttacttcttct ggtttctaatgatattccaggaaaggagcctaactagagtcagtagaaggccatcctaggaaacaatcccatgagtgatgtttcatgaattatcttcgtgaaccggagcaagtcgctttgccttcctgaaccttctctgagatggaagggtgatctgaaaatatgcctcccgggtaaacctcagccctggctggcagatctgg ctggctaatgggactccccttttcctctccatggctcctgtgcctccccctcaagactgtgtgtattccaagaggactacattcccaggtggggtcgtattgttcttataatgaaggcttacagaatttcctcgaatgttgcatctagttttagcgttctgaatctgtgaatcacaaattagaattggtgcctttggaagag gagatgggacatgaagcctgctccaccttctgataatgccctaaagagcagctcaaaatacagctaaaagaacccagaaaacaaaataggtaagtcttgaagagagatcaacattaggaacacaatattcactgaatataattctgtggtctgttttatactttatttttgcattcactcctagtatttaatttatgtatcaatttctgagaatt ggtttccaaaaaagtcattatccagtgaactctattagtcactaaagctttataacaaataaattaacaaaacattgttaccctaagaaggaatagcctcttgtcacttgttgccccttggctctcaaaactggaaagtccacatgcttatagtggttatgatagtttatcaaactgaagaagaagcctgaacaacact ccagtgattccagacacaagtagataatctattcagccccatgtagtttcagagagtaaacgaagataaggaaatttttttcttctcctcagggcagagatgtttttcagttcttcttcttttttttttttaatcttagcacttgatattcccttgggaagtgttggttacaacaaaacactatttttagaatcca agtatctgaaattaatcaaagatagattagcatgccagtgtcccctttcctcttttgttataaaagagcaatagtcattttactattgctaaatgtcatttacatttagtcatttactaaaaattagtaaaaaatttcttaatataagcataaagttacaaaggcaaagagaagatgtactggaaggtaaataggcgtggcaga ggtttgaaaggctgttaggcactggattataatccttctagggtatggacaacatctgggtgttattttctctttcatagtcagttcagtactcactgtgcatgagaggtgcttgagtgagtctttgttagataaatgaaaagcaaattaatccacttttcagagctctcttcggagacctgagcctgaacaagacaatt ctctaaaagacagaatatttgggaatgaaaaaataggacattagaagatgctgtttgttactttttaaatcccttttcatcctggctacgctaactctacccctcaagggcaggagaacagaggacgagatggctccaggcaagttgtgagctgtgtggcatcagcatctgtctatagtggtgatgctggtgctgacatgtggt ggtttattaatctatttgggccccaatatctgcatgtgtaaaagggaatgatgataatgagtattttccaaaaatatggttgaaggattaactgtatttgagatagagtaggcctcagttcgtgtcagctattagccttagtctgtctacaggaaccagaagtttcaggttgggagaagcctggccttttagtga gaagctttcttgtggtgggacaagatgttggtgtgagaccagaaactcagcctctcaggagcagaaaaagaggggactcgatgttctgctggtgattctgagatgagtgttaacatcagatgtggctgttagtccccagaaggacaaatataaatgtttccttcatgctcttctttgtttttgtttt tggtttttgttccttctgctgataaatattcttactcagaggttattactgcaaataataataaaaaatactggtaaaccaggcctctgatgtgttattgttaaaaaaaatcaatgttttatcacagtctgagtacagacttggcttattgttctgaaagaattctgcatggcagggcaggggctgagcaaaagat taatgttcaaaggatttattagtgttgactgtgagtatctgtttgtgtttgtctgtaaagttggcttggtagtcagaatgtatgggaaaaccttctaagagtcatatggcagcttttcaaaaactctgtagagctgggtacacaatttccctggactcactcccatctaagcagcctctatcagagggaccagagcag ctgtaagatacctgttcaggacagatgtgcataggcctgaagcaggatactgcatcccaagcaaccacagactgccaagaaagtttcctgtcatggcacaacttaagctgaaaaaaaagcatcacagcctgagctggaaggggtccgagagctgctatcatccaatcatccaatcccttcattttctagttgaggaatcaaatctacacatct ctagcgacctgcccaagataatgcagctaattagggtcagagttaagatttgagacagaatacagaaatttttttttaaagccttctgtactgacatttgttttcattaaatttgataccacttctatcctttgtcaacttttatcgctgagaaataattaatgttgcaataagctggaatcaaagaaattggtgagagaaattgt gctgacatgtttggggttgtctttgtattctctgaaaccacENST00000658243.1 (LINC00958) length = 1814 (SEQ ID NO. 3)

ctctctctctctcctgctgcattgtgaagaaactgcttgcttccctctcaccctctgcag

tttcctgaggcctccccagccatgcggaacgggagaaagtttaagctctcctcactatga

gttttcaagtataaaagactttttcttccacgattttgagaacaactgaggactcttgtg

accaggacaacagggaagcttgcagcaagatagctccaggttggattcatgcttcgcacc

ccaagggctgccagccagagaggggagaagcaatcactcctgcagtttctgaacactac

acagacgccaggtagcttcttcaggagaacagccctctgaggaggcaggaagaggaggct

tatctttcagcagccggagctgctgagatctctgggcagattaagctctctctaatggat

gggctccagcctggcacattcagtggagagggatccactcatccatcatcaacataatat

ggtcctccctgcacttcacagtgtcctcttgctattgaaaaggcttttttgccttctcaa

gtttctttgtcaacagtctacaggaagaagctcaggccgccaccggcagagaaccttcca

ctggccactcagcgaaaggcagctgattcagcacctggagaagaccggcaacagggctga

gtttccttgcactgcagaggagggggagaggcaaggagtggacatgagcacggaggatcca

gggcactcccttgatggtgatgccctcgaagggctgccccgtgcccattgtacagaatgt

ggattccaccccacgccatgagagctctcgcgcaggcccagaccactttataagccatat

ttcccaccaattcctgtttctggatctgtcaggttatttcatatttctaggcccttagct

ttgcttcctcttccctgaacgccctttgcccacatcatccccagctctccttctccctcctg

ccttctgtcttctcttcctctcatttcttgtcacccaggctggagtccagtggcgtgatc

acggctcactacagccttgaactcccgggctcaggtgatcctcccacctgagcctcccga

gtagctgggactacagatgcacaccaccatgccagctaatttttaaaactgtttttaggg

atggggtcatcttcctgtgttgcccaggcttgtctcaaactcctgggctcaagtgatcct

ctcacctcagccttccaaagtgctgggattacaggcatgagccacaacatccagcccctt

ttctcttttcttacccttctttcctattttcttttccattttctttccctcccttcttct

ttctttcctaactattaaggagtagattgaattcaaggtctttatgtgtgtcagtttttg

ttttccaacaaatatttcttaaaaaccaaccattgaaacgtaatggtaaccactggcccc

tgtctccacctccaccacctaagaagccccaaatccagatgtgtccattaaaatcagtcca

gatcttctttaccaagccactagatgtcatattaatttcacagcagaatagggaagccca

tgccggagctgaaaacctgcaacaacaaaaaagcatctaaatactgcaaaaaaaaaaaaaa

aaaaagaggggagagaggcatcttaatatttgtccttaacacatcataccagacttctgct

tccaggaatgactggatgtggtttttctaggtggacataatacaaaatgcagctacctga

aaacaatggaaagtggacagagcaggcagattctggtgggaagtccacgcaataaagaaa

agatgttacaggaa

2. Isolation and identification of exosomes: Take 1.5ml of serum from patients with esophageal squamous cell carcinoma and healthy controls, centrifuge at 2000×g, 4°C for 30 minutes, and then centrifuge the supernatant at 10,000×g, 4°C for 45 minutes. The supernatant was then collected and filtered through a membrane with a pore size of 0.22 μm. The filtrate was collected and ultracentrifuged at 10000×g for 70 min at 4°C. Resuspend the pellet in 10 ml of pre-chilled 1× PBS and repeat the ultracentrifugation. Finally, resuspend the pellet in 1.5 ml of pre-chilled 1× PBS. Isolated exosomes were characterized using transmission electron microscopy (TEM). Exosome concentration-size assay (NTA) was performed by Zeta View (Particle Metrix Ltd. Germany).

3. Exosome RNA extraction: use the total exosome RNA isolation kit from Invitrogen (Cat. No.: 4478545), extract exosome RNA according to the protocol provided by the kit, and synthesize cDNA using the PrimeScriptRT Master Mix kit from TAKARA. The synthesis protocol follows Kit instructions were carried out. The target lncRNA was quantified using the TAKARA SYBRgreen kit, and the calculation formula of the result was: 2 ^-△t (t represents the difference between the number of cycles required for the target gene and the reference gene to reach the threshold, using β-actin as the internal reference gene).

Quantitative detection primer sequences are as follows:

AC098818.2F:GCAGGACGGAACTGGAGTGGAT (SEQ ID NO.4)

R: GTGGTACAGCAGGCGTGAGATG (SEQ ID NO. 5)

LINC00958 F: CTACACAGACGCCAGGTAGC (SEQ ID NO. 6)

R: GCTCCGGCTGCTGAAAGATA (SEQ ID NO. 7)

RASSF8-AS1 F:CTCCACCTTTGCCTTCCACCAT (SEQ ID NO.8)

R: GCCTCAAGCAATCCTTCCACCT (SEQ ID NO. 9)

β-ACTINF:GTCATTCCAAATATGAGATGCGT (SEQ ID NO.10)

R: GCATTACATAATTTACACGAAAGCA (SEQ ID NO. 11)

(1) Reverse transcription reaction

1) Remove genomic DNA

reaction system

Reaction conditions: 37°C, 4min

2) Reverse transcription to generate cDNA template reaction system

Reaction conditions: 37°C for 15 minutes, 85°C for 5 seconds

(2) qPCR

qPCR reaction uses β-actin as an internal reference gene

reaction system

qRT-PCR program settings

4. Stability detection of exosomal lncRNAs: Serum exosomes samples from 15 patients with esophageal squamous cell carcinoma were treated under different conditions to compare whether the expression levels of candidate lncRNAs changed significantly under different treatment conditions. Treatment conditions include: 1) treatment at different ambient temperatures (Fig. 3A), 2) treatment with RNase (Fig. 3B); 3) samples subjected to repeated freezing and thawing (Fig. 3C); 3) samples treated with acid/alkali (HCl/ NaOH) treatment (Figure 3D).

5. Comparison of expression levels of exosomal lncRNAs before and after surgery: 20 patients with esophageal squamous cell carcinoma were selected, and their serum was extracted before surgery and 1 month after surgery. Exosomal RNA was extracted according to the above procedure, and the expression levels of candidate lncRNAs were quantified The differences in the expression levels of patients before and after surgery were compared. The results are shown in Figure 4, the expression levels of the three candidate lncRNAs were down-regulated to varying degrees after surgery.

6. Establish a diagnostic model (training set stage): In the serum of 40 patients with esophageal squamous cell carcinoma and 40 healthy controls, the above method was used to isolate serum exosomes, extract exosomal RNA, reverse transcribe to generate cDNA, and perform qRT- PCR method was used to quantitatively detect the expression of each lncRNA, and ROC curve analysis was used to establish a diagnostic model for each marker and a combined diagnostic model for multiple markers. In addition, according to the detected serum tumor marker data of each patient, ROC analysis was used to calculate the AUC value of each tumor marker independently for the diagnosis of esophageal squamous cell carcinoma, and the AUC value of the combined application of multiple tumor markers for the diagnosis of esophageal squamous cell carcinoma . A single index weight coefficient is given through logistic regression analysis, and a diagnostic model is constructed for publicity. The calculated diagnostic models are Index I=EXP AC098818.2*0.74429+EXP LINC00958*1.24591+EXPRASSF8-AS1*1.27292

Index II＝EXP CA125*9.66194+EXP CA153*0.047267+EXP CA199*0.033176+EXPCY211*0.0032795+EXP NSE*0.025847+EXP SCC*(-112.09409)

Among them, Index I is a diagnostic model based on exosomal lncRNA calculations, and Index II is a diagnostic model based on tumor markers. EXP: EXPRESSION, expression level.

According to single index and combined index, the AUC values of the two diagnostic models were calculated to be 0.84 and 0.63, and the cut-off values were 22.84 and 2.628, respectively. The above experimental results show that the diagnostic value of the combined diagnosis model of lncRNA markers is better than that of traditional tumor markers.

6. Verify the diagnostic model (validation set stage):

In 140 patients with esophageal squamous cell carcinoma and 140 healthy controls, the total RNA of serum exosomes was extracted, and the expression level of the target lncRNA was quantitatively detected. The AUC value and 95%CI of traditional tumor markers combined diagnosis of esophageal squamous cell carcinoma, and compared the advantages and disadvantages of Index I and Index II. The experimental results show that in the validation set, Index I still shows better diagnostic value than Index II.

The embodiments of this specific implementation mode are all preferred embodiments of the present invention, and are not intended to limit the protection scope of the present invention, so: all equivalent changes made according to the structure, shape and principle of the present invention should be covered by within the protection scope of the present invention.

Claims (3)

1. An exosome lncRNA marker set for esophageal squamous carcinoma diagnosis, characterized in that: the exosome lncRNA marker group consists of lncRNA AC098818.2, RASSF8-AS1 and LINC00958.

2. A diagnostic model based on the exosome lncRNA marker set of claim 1, characterized in that: the diagnostic model is EXP AC098818.2 x 0.74429+EXP LINC00958*1.24591+EXP RASSF8-AS1 x 1.27292.

3. Use of the exosome lncRNA marker set of claim 1 in the preparation of a product for diagnosing esophageal squamous carcinoma.

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