KR102136747B1 - Diagnostic Biomarker For Prognosis of Intestinal Type Gastric Cancer - Google Patents

Abstract

The present invention provides a composition for predicting the prognosis of intestinal type gastric cancer comprising an agent for measuring a gene expression level of VGLL1 (Vestigial Like Family Member 1), a kit comprising the same, and information on predicting the prognosis of intestinal gastric cancer As a method, as the expression level of the gene has a close correlation with the overall survival rate and the recurrence-free survival rate of patients with long-term gastric cancer, by measuring the expression level of the gene, the prognosis of patients with long-term gastric cancer is predicted to be effective in diagnosis and treatment. Can be used.

Description

Diagnostic Biomarker For Prognosis of Intestinal Type Gastric Cancer}

The present invention relates to a composition for predicting prognosis that can predict the prognosis of intestinal type gastric cancer, a kit comprising the same, and a method for providing information related to prognosis prediction.

Stomach cancer is the number one cancer in the world with the highest incidence of Koreans, and is one of the most important diseases for prevention, treatment, and prognosis. Stomach cancer can be divided into an intestinal type and a diffuse type when classified according to pathological and histologically based Lauren classification. In the case of long stomach cancer, cancer cells form a mass and proliferate, and the infiltration rate tends to be slow. Diffuse gastric cancer proliferates in a shape that spreads under the mucous membrane of the stomach and has the characteristic of cancer cells infiltrating into the stomach wall. The incidence is high in young people in their 20s and 30s.

In the case of the long form, the prognosis of treatment may be good because it can be found at a relatively early stage compared to the sub form, but it may be necessary to detect the onset early because the patient may die in terms of survival rate and recurrence rate. Among the long-term patients, if a patient with a poor prognosis or a patient with a high probability of recurrence can be identified and predicted in advance, there is a possibility that a treatment strategy and a survival rate can be increased by establishing a future treatment strategy.

Since the expression of various genes is abnormally controlled in the onset of cancer, in predicting the diagnosis or prognosis of cancer onset, the expression level of a gene associated with a specific cancer is compared with the expression level in normal cells. Can. By finding biomarkers that can be used as indicators for measuring the presence, severity, and prognosis of a specific disease, and discovering compositions and methods that can analyze them, the success rate and viability in the treatment of diseases To help reduce the pain and cost of patients.

HER2 (Human Epidermal Growth Factor Receptor 2 ) gene, when overexpressed genes with the ability to engage in regulatory mechanisms related to the division of cells, or activation and protein molecules have been known to cause cancer, in the diagnosis of various cancers HER2 A method for measuring and confirming the expression level of a gene is presented. Even in patients with long stomach gastric cancer, the expression level of HER2 gene is high, but according to the analysis, there is a disadvantage in that the malignant association with prognosis is unclear. Therefore, in order to predict the prognosis of patients with gastrointestinal cancer, more accurate diagnostic markers are needed.

An object of the present invention is to provide information for predicting the prognosis of a gastric cancer patient, among others, a composition and kit capable of predicting the prognosis of a gastric cancer patient, among others, an intestinal type, with a higher accuracy.

In order to achieve the above object, one aspect of the present invention provides a composition for predicting gastric cancer prognosis comprising an agent that measures the expression level of the VGLL1 gene.

In addition, another aspect of the present invention provides a kit for predicting gastric cancer prognosis comprising the composition.

In addition, another aspect of the present invention comprises the steps of measuring the amount of mRNA or VGLL1 gene mRNA present in a biological sample from a patient with long stomach gastric cancer and comparing it with the amount measured from a sample of a normal individual. Provides a way to provide information.

According to the present invention there is found that the expression level of VGLL1 gene shown to be highly correlated with the prognosis of the intestinal gastric cancer patients, there is an effect that can more accurately predict the prognosis of the intestinal gastric cancer by measuring the expression level of VGLL1 gene.

However, the effects of the present invention are not limited to the above-mentioned effects, and other effects not mentioned will be clearly understood by those skilled in the art from the following description.

1 is a graph analyzing the difference in the expression level of the VGLL1 gene in long gastric cancer and diffuse gastric cancer based on clinical data of a total of 436 patients with long and short gastric cancer.
A and B of FIG. 2 are based on clinical data of gastric cancer patients with 239 long (A) and 197 less (B) patients with high expression of the VGLL1 gene ( VGLL1 -high) and low expression ( VGLL1- This graph shows the analysis of the difference between overall survival (OS) and recurrence-free survival (RFS).
FIG. 3A shows the correlation between the expression level of the HER2 gene ( ERBB2 gene) and the expression level of the VGLL1 gene in tissues of patients with long stomach gastric cancer. B and C of Figure 3 is a graph analyzing the expression level of the HER2 gene (ERBB2 gene) based on clinical data of 239 long (B) and 197 less (C) gastric cancer patients. FIG. 3B shows the difference in overall survival and recurrence-free survival between patients with high expression of the HER2 gene ( ERBB2 gene) ( ERBB2 -high) and patients with low expression ( ERBB2 -low) among patients with long stomach gastric cancer. C represents the difference between the overall survival rate and the recurrence-free survival rate among patients with diffuse gastric cancer who have high expression of the HER2 gene ( ERBB2 gene) ( ERBB2 -high) and low expression ( ERBB2 -low).
FIG. 4 is a graph showing that the prognosis is poor in a patient group with high expression of both genes through a correlation analysis of the overall survival rate and recurrence-free survival rate of the gastrointestinal cancer patient and the expression levels of the VGLL1 gene and the HER2 gene ( ERBB2 gene).
VGLL1(-), ERBB2(-): Patients with low expression levels of both VGLL1 and HER2 genes ( ERBB2 gene);
VGLL1(-), ERBB2(+): Patients with low expression levels of the VGLL1 gene and high expression levels of the HER2 gene ( ERBB2 gene);
VGLL1(+), ERBB2(-): Patients with high expression levels of the VGLL1 gene and low expression levels of the HER2 gene ( ERBB2 gene);
VGLL1(+), ERBB2(+): Patients with high expression levels of both VGLL1 and HER2 genes ( ERBB2 gene).
FIG. 5 confirms whether there are other genes that specifically express high or specifically low levels of expression in long gastric cancer tissues with high expression level of the VGLL1 gene, and the top 21 predicted to be related to the prognosis of the patient. This is a table for selecting genes.
6A is a graph based on the expression level of the top 21 genes of FIG. 5, and the risk score of patients with long stomach gastric cancer is classified into a high-risk group and a low-risk group, and FIG. 6B analyzes the overall survival rate between the two groups It is a graph.

First, the terms used in the present invention are defined.

In the present invention, "prognosis" refers to the prospect of a future medical condition (eg, overall survival rate, survival rate without recurrence, etc.) for a patient's disease, and the degree of disease (eg, recovery of disease) Etc.), the degree of improvement of the disease (e.g., tumor regression, etc.) or a positive prognosis including stabilization of the disease, disease recurrence, disease progression or fatality (e.g., tumor Growth, metastasis, drug resistance, etc.).

In the present invention, "prediction" means to predict and predict the future medical condition of the patient's disease, and the course of the patient's disease (eg, degree of disease, degree of improvement, stabilization, relapse, progression) Degree, fatality, exacerbation, etc.).

In the present invention, the "biological sample" may be, but is not limited to, tissue, cells, urine, saliva, whole blood, serum, plasma, and cerebrospinal fluid obtained from patients with long stomach gastric cancer or normal individuals.

Hereinafter, the present invention will be described in detail.

One. Compositions and kits for predicting long-term gastric cancer prognosis

The present invention provides a composition for predicting long-term gastric cancer prognosis, and a kit for predicting long-term gastric cancer prognosis comprising the composition.

The composition for predicting prolonged gastric cancer prognosis of the present invention includes an agent that measures the expression level of the VGLL1 (Vestigial Like Family Member 1) gene.

The gastrointestinal cancer is distinguished from diffuse type gastric cancer when gastric cancer is classified according to Lauren's classification based on histopathological and histological characteristics. Cancer cells form a lump and proliferate by flocking in one place. It means stomach cancer.

The VGLL1 (Vestigial Like Family Member 1) gene is a gene encoding the Vestigial Like Family Member 1 protein. The protein expressed from the VGLL1 gene interacts with and binds to the TEA domain family of transcription enhancer factor (TEF) in mammals, thereby acting as a coactivator to increase the expression of the oncogene. It is known to have the characteristic of inducing cell proliferation accordingly. The nucleotide sequence and amino acid sequence of the protein expressed from the gene of VGLL1 VGLL1 gene can be obtained from the published databases or literature, such as the NCBI GenBank.

The agent for measuring the expression level of the VGLL1 gene may be an agent for measuring the amount of mRNA of the gene or the amount of protein expressed from the gene.

An agent that measures the amount of the mRNA of the VGLL1 gene refers to an agent that specifically binds to the gene mRNA and allows recognition or amplification of the amount. As a specific example, it may be a primer pair or a probe that specifically binds to the nucleotide sequence of cDNA produced by reverse transcription of the mRNA or mRNA.

The primer is a short-length nucleic acid sequence having a free 3'terminal hydroxyl group and forms a base pair with a complementary template strand to start the nucleic acid polymerase when replicating and amplifying the template strand. It is a nucleic acid sequence that serves to provide a point. In addition, the probe refers to a nucleic acid fragment having a length of several bases to hundreds of bases consisting of a sequence capable of specific binding with mRNA or cDNA.

The amount in which the mRNA of the gene is present includes PCR, RT-PCR, competitive RT-PCR (competitive RT-PCR), and real-time RT-PCR (RT-PCR) using sense and antisense primers of the gene sequence. It can be measured by performing the method, and performs a method such as Northern blot (Northern blot), microarray (Microarray) using a probe having a sequence capable of specific binding to the mRNA or cDNA made by reverse transcription of the gene It can be measured by, and can be measured by methods such as RNase protection assay, sequencing, etc., but is not limited thereto, and any preparation necessary to use any method known to those skilled in the art is possible. .

An agent that measures the amount of the protein expressed from the VGLL1 gene exists refers to an agent that specifically binds and recognizes the protein. As a specific example, it may be an antibody or aptamer that specifically binds to the protein.

The antibody immunologically refers to an immunoglobulin molecule having a specific binding reactivity by specifically binding with an epitope of the protein, a monoclonal antibody, a polyclonal antibody, an antibody having a full-length chain structure, at least antigen binding function It may include both antibodies and recombinant antibodies of functional fragments. The aptamer refers to a single-stranded nucleic acid molecule having a characteristic capable of forming a specific binding by targeting the protein and forming a stable tertiary structure, using SELEX (Systematic Evolution of Ligands by Exponential enrichment) technology, etc. By aptamer having specificity to the protein can be synthesized.

The amount of protein expressed from the gene is Western blot, protein microarray (protein chip), ELISA (Enzyme Linked Immunosorbent Assay), 2-dimensional electrophoresis (2-dimentional electrophoresis) using the antibody or aptamer. Immunohistochemistry (IHC), immunofluorescence, co-immunoprecipitation assay, FACS (Fluorescence activated cell sorter), radioimmunoassay (RIA), radioimmunodiffusion, It can be measured by a method such as MALDI-TOF analysis (Matrix Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry), but is not limited thereto, and any formulation necessary to use any method known to those skilled in the art is possible.

The composition comprising an agent for measuring the expression level of the VGLL1 gene can be used as a predictive purpose for the prognosis of a patient with long stomach gastric cancer. As a result of measuring the expression level of the VGLL1 gene, when the expression level of the gene is high, it is possible to predict that the prognosis of patients with long stomach gastric cancer will be poor, and strategies and measures for treatment can be established. In this connection, the specific examples 1 and 2 the expression level of VGLL1 gene overall survival in gastric cancer appears higher from the intestinal gastric cancer compared to the diffuse type gastric cancer patients among patients, intestinal amount of expression of VGLL1 gene high gastric cancer of the present invention ( Overall survival (OS) and recurrence-free survival (RFS) were found to be higher than those with low gastrointestinal cancer, where the expression level of the VGLL1 gene was low (see FIGS. 1 and 2).

The composition for predicting prolonged gastric cancer prognosis may further include an agent that measures the expression level of a gene of HER2 (Human Epidermal Growth Factor Receptor 2).

The HER2 ( Human Epidermal Growth Factor Receptor 2) gene is a gene that encodes the Human Epidermal Growth Factor Receptor 2 protein, and is named ERBB2 in humans. It is a gene that can cause cancer when the number of genes corresponding to a proto-oncogene is amplified, overexpressed, or when an expressed protein is overactive. It functions as a receptor tyrosine kinase bound to the membrane and is involved in regulatory mechanisms related to cell division. The nucleotide sequence and amino acid sequence of the protein expressed from the gene of HER2 HER2 gene can be obtained from the published databases or literature, such as the NCBI GenBank.

The agent for measuring the expression level of the HER2 gene includes an agent for measuring the amount of mRNA of the gene or the amount of protein expressed from the gene.

The HER2 agent measuring the amount of mRNA the presence of the gene or the kind of formulation that measures the amount of the expressed protein from the HER2 gene present with regard to the preparation for measuring the amount of the mRNA or protein of said VGLL1 gene present Same as described.

In addition to the expression level of VGLL1 gene, by measuring with the expression level of the HER2 gene, in the case of measuring only the expression level of VGLL1 gene or HER2 predict more prognosis of intestinal gastric cancer with high accuracy as compared with the case of measuring only the expression level of the gene It has the effect. In this regard, in the specific embodiment 3 of the present invention, in the case of a long stomach gastric cancer patient having high expression levels of both the VGLL1 gene and the HER2 gene, there is no overall survival rate and recurrence compared to a patient in which both genes have high expression levels or both genes have low expression levels. It was confirmed that the survival rate was significantly low (see FIG. 4).

In addition, the composition for predicting prolonged gastric cancer prognosis is MMP9 ( Matrix Metallopeptidase 9 ), WNT7A (Wnt Family Member 7A), RTN3 (Reticulon 3), EDA (Ectodysplasin A), ROPN1B (Rhophilin Associated Tail Protein 1B), WIPF3 (WAS/ WASL Interacting Protein Family Member 3), GCNT2 (Glucosaminyl (N-Acetyl) Transferase 2), TM7SF3 (Transmembrane 7 Superfamily Member 3), CRABP2 (Cellular Retinoic Acid Binding Protein 2), KLK7 ( Kallikrein Related Peptidase 7), DEFB1 (Defensin Beta 1), RPS6 (Ribosomal Protein S6), HNRPM (Heterogeneous Nuclear Ribonucleoprotein M), AURKAIP1 (Aurora Kinase A Interacting Protein 1), NASP (Nuclear Autoantigenic Sperm Protein), CPSF2 (Cleavage And Polyadenylation Specific Factor 2), GFI1 (Growth) Expression of one or more genes selected from the group consisting of Factor Independent 1 Transcriptional Repressor), SNORD31 (Small Nucleolar RNA, C/D Box 31), RPSA (Ribosomal Protein SA), ELAVL1 (ELAV Like RNA Binding Protein 1) and LRRC48 The agent for measuring the amount may further include an agent for measuring the expression level of one or more genes selected from the group. The amount of mRNA of the gene is present or one or more genes selected from the group. It includes an agent that measures the amount of protein present therefrom.

The type of the agent that measures the amount of mRNA present in one or more genes selected from the group or the type of the agent that measures the amount of proteins expressed from one or more genes selected from the group is the mRNA of the VGLL1 gene or This is the same as described for the agent measuring the amount of protein present.

By measuring the expression levels of one or more genes selected from the group together with the expression levels of the VGLL1 and HER2 genes, there is an effect that can further increase the accuracy in the purpose and use of the present invention for predicting the prognosis of patients with long stomach gastric cancer. . In this regard, in Example 5 of the present invention, a long-term gastric cancer patient was classified into a high-risk group and a low-risk group based on the expression level of genes in the group, and the correlation between the overall survival rate and the overall survival rate of the high-risk group was low-risk group. It was confirmed that it was significantly lower than (see FIG. 6).

The composition can be provided in the form of a kit.

The kit may include a composition, solution or device comprising one or more other components suitable for measuring the amount of mRNA present in the VGLL1 gene, HER2 gene and one or more genes selected from the gene group.

The kit may be an RT-PCR kit, a DNA chip kit, but is not limited to this, and any kit necessary to use any method known to those skilled in the art is possible.

Specifically, the kit may be a kit including essential components necessary for performing RT-PCR. For example, in addition to a pair of primers that can specifically bind to the mRNA of the gene, test tubes or other suitable containers, enzymes such as Taq-Polymerase and Reverse transcriptase, reaction buffer solutions (pH and magnesium concentrations may vary), deoxynucleotide (dNTP), DNase and RNase inhibitors, DEPC-water (DEPC-water), sterile water, and the like. In addition, when quantifying, it may include a primer pair capable of specifically binding to a gene used as a control.

As another specific example, the kit may be a kit including essential components necessary to perform a DNA chip. It may include reagents, enzymes, agents, and the like for preparing a glass substrate and a fluorescent label probe to which an oligonucleotide capable of specifically binding to the mRNA or reverse transcribed cDNA of the gene is attached.

The kit may comprise a composition, solution or device comprising one or more other components suitable for measuring the amount of the VGLL1 gene, the HER2 gene and the protein expressed from one or more genes selected from the gene group. have.

The kit may be an ELISA kit, a protein chip kit, but is not limited thereto, and any kit necessary to use any method known to those skilled in the art may be used.

Specifically, the kit may be a kit including essential components necessary for performing ELISA. For example, it may include a substrate for immunological detection of an antibody, an appropriate reaction buffer, and a reagent capable of detecting the formation of an antigen-antibody complex using an antibody capable of specifically binding to a protein expressed from the gene. It may include, for example, a secondary antibody labeled with an enzyme, a fluorescent substance, etc., which mediate a chromogenic reaction, and a chromogenic substrate. Examples of the enzyme that mediates the color reaction may include alkaline phosphatase, acid phosphatase, peroxidase, and examples of the fluorescent material include colloidal gold and flu. Oresinecarboxylic acid (FCA), fluorescein isothiocyanate (FITC), RITC (Rhodamine-B-isothiocyanate), fluorescein thiourea (FTH), 7-acetoxycoumarin-3-yl, fluorescein- 5-yl, fluorescein-6-yl, 2',7'-dichlorofluoresin-5-yl, 2',7'-dichlorofluoresin-6-yl, dihydrotetramethylrosamin-4 -Yl, tetramethylrodamine-5-yl, tetramethylrodamine-6-yl,4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-indacene-3 -Ethyl or 4,4-difluoro-5,7-diphenyl-4-bora-3a,4a-diaza-s-indacene-3-ethyl and the like. In addition, when quantifying, it may include an antibody that specifically binds to a protein expressed from a gene used as a control.

As another specific example, the kit may be a kit including essential components necessary to perform a protein chip. For example, one or more antibodies capable of specifically binding to a protein expressed from the gene may be immobilized and arranged at a predetermined position on a substrate. In addition, when quantifying, it may include an antibody that specifically binds to a protein expressed from a gene used as a control.

2. How to provide information to predict the prognosis of patients with gastrointestinal cancer

The present invention provides a method for providing information for predicting the prognosis of a patient with long stomach gastric cancer.

The method of the present invention comprises the steps of measuring the expression level of the VGLL1 gene from a biological sample of an intestinal gastric cancer patient; And comparing the expression level of the measured gene with the expression level in a normal individual.

In the step of measuring the expression level of the VGLL1 gene, the expression level of the HER2 gene can be further measured, wherein MMP9 , WNT7A, RTN3, EDA, ROPN1B, WIPF3, GCNT2, TM7SF3 CRABP2, KLK7, DEFB1, RPS6, HNRPM, The expression level of one or more genes selected from the group consisting of AURKAIP1, NASP, CPSF2, GFI1, SNORD31, RPSA, ELAVL1, and LRRC48 can be further measured.

The step of measuring the expression level of the gene may be to measure the amount of mRNA of the gene, or to measure the amount of protein expressed from the gene.

The step of comparing the expression level in the normal human subject includes comparing the expression level of the measured gene with the expression level data of the gene obtained from a person who does not have an intestinal gastric cancer disease. In addition, the expression level data of the gene obtained from a person who does not have the intestinal gastric cancer disease includes that obtained by measuring the amount of mRNA of the gene or the amount of protein expressed from the gene.

The step of measuring the amount of mRNA present in the gene refers to a step of specifically binding to the gene mRNA and recognizing it or using an agent capable of amplifying the amount. As a specific example, it may be measured using a primer pair or a probe that specifically binds to the nucleotide sequence of cDNA produced by reverse transcription of the mRNA or mRNA, PCR, RT-PCR, Competitive RT-PCR (Competitive RT-PCR) , Real-time RT-PCR (Real-time RT-PCR), Northern blot (Northern blot), microarray (Microarray), RNase protection assay (RNase protection assay), can be measured by methods such as sequencing, etc. However, the present invention is not limited thereto, and any measurement method known to those skilled in the art can be used.

The step of measuring the amount of the protein expressed from the gene refers to the step of measuring using an agent that specifically binds and recognizes the protein. As a specific example, it may be measured by using an antibody or aptamer that specifically binds to the protein, and Western blot, protein microarray (protein chip), ELISA (Enzyme Linked Immunosorbent Assay), 2-dimensional electrophoresis, immunohistochemistry (IHC), immunofluorescence, co-immunoprecipitation assay, FACS (Fluorescence activated cell sorter), radioimmunoassay , RIA), radioimmunodiffusion, MALDI-TOF analysis (Matrix Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry), but is not limited thereto, and any method known to a person skilled in the art can be used. It is possible to measure.

The method may further include a step of predicting that if the expression level of the VGLL1 gene or the expression level of the VGLL1 and HER2 genes is higher than that of a normal individual, the prognosis of a patient with long stomach gastric cancer will be poor.

Along with the expression level of the gene or the expression level of the VGLL1 and HER2 genes, MMP9 , WNT7A, RTN3, EDA, ROPN1B, WIPF3, GCNT2, TM7SF3 CRABP2, KLK7, DEFB1, RPS6, HNRPM, AURKAIP1, NASP, CPSF2, GFI1, SNORD When the expression level of one or more genes selected from the group consisting of RPSA, ELAVL1 and LRRC48 is further measured, in predicting the prognosis of the patient with gastrointestinal cancer, based on the expression level of one or more genes selected from the group By calculating the risk score, if the risk score is analyzed to be higher than the risk score in the normal subjects, it can be predicted that the prognosis of patients with long stomach gastric cancer will be worse with higher reliability.

The risk score is calculated by applying a Cox proportional hazard model to the gene expression level of the group, and calculating the hazard ratio of each gene, and the expression value and risk ratio coefficient of each gene ratio coefficient). Specifically, the risk score can be calculated using the following equation.

Risk score of long stomach gastric cancer = ∑ (gene expression level value) X (gene risk ratio coefficient)

Hereinafter, the present invention will be described in detail by examples and experimental examples.

However, the following Examples and Experimental Examples specifically illustrate the present invention, and the contents of the present invention are not limited by the following Examples and Experimental Examples.

In gastric cancer tissue VGLL1 Analysis of gene expression level

Based on the clinical data of 436 gastric cancer patients received from Severance Hospital, Yonsei University, the difference in expression levels of VGLL1 gene in two types of gastric cancer patients according to histological classification was analyzed. When the VGLL1 expression levels of two types of gastric cancer patients were compared by categorizing them as intestinal type gastric cancer and diffuse gastric cancer, the expression level of the VGLL1 gene was significantly higher in gastric cancer than in gastric cancer. It was confirmed (see Figure 1). Through this, it was confirmed that, among the two types of gastric cancer, the expression of the long gastric cancer and the VGLL1 gene is related.

In the tissues of patients with gastric cancer VGLL1 Analysis of correlation between genes and survival rate

Survival rates were analyzed based on clinical data from 436 patients with long and short gastric cancer provided by Yonsei University Severance Hospital. As a result, when the VGLL1 gene expression in the tissues of patients with long stomach gastric cancer is low, the overall survival rate (OV) is compared with the long stomach gastric cancer patient ( VGLL1 -low) and the long gastric cancer patient with high VGLL1 gene expression ( VGLL1 -high). And recurrence-free survival (RFS) was found to be lower in patients with long stomach gastric cancer ( VGLL1- high) with high expression of the VGLL1 gene (see A in FIG. 2).

However, in patients with diffuse gastric cancer, there is no significant difference in overall survival or recurrence-free survival in patients with low expression levels of VGLL1 gene in patients with high expression levels of VGLL1 gene, and thus does not show a significant correlation with the expression level of VGLL1 gene. It was confirmed (see B in Figure 2).

Through the above results, it was found that the expression level of the VGLL1 gene is closely related to the overall survival rate and the survival rate without relapse in the case of patients with long stomach gastric cancer, but not in the case of patients with gastrointestinal cancer. Therefore, when the expression level of the VGLL1 gene in the tissue of a patient with long stomach gastric cancer is measured high, the prognosis may be poor because the overall survival rate and the survival rate without recurrence are low.

In patients with long stomach gastric cancer VGLL1 Gene expression and HER2 Check the correlation between gene expression

Gene expression was analyzed based on clinical data from a total of 436 patients with long and short gastric cancer provided by Severance Hospital, Yonsei University. When comparing the expression levels of the VGLL1 gene and the HER2 gene expressed in the tissues of patients with long stomach gastric cancer, it was confirmed that there is a certain level of correlation between the expression levels of the two genes (see FIG. 3A). It was confirmed that the expression level of the HER2 gene ( ERBB2 gene) in the tissues of patients with long stomach gastric cancer has a certain correlation with the overall survival rate and the recurrence-free survival rate of patients with long stomach cancer, similar to the expression level of the VGLL1 gene (see FIG. 3B ). ). However, the expression level of the HER2 gene ( ERBB2 gene) in the tissue of patients with diffuse gastric cancer was not correlated with the overall survival rate (see FIG. 3C).

Furthermore, the survival rate was analyzed by dividing the VGLL1 gene and the HER2 gene in the case of gastrointestinal cancer patients with high expression, only one overexpression, or both genes not overexpressing. As a result, it was confirmed that in the case of a long stomach cancer patient with high expression of both genes, the overall survival rate and the recurrence-free survival rate were significantly lower than those of the long stomach gastric cancer patients (see FIG. 4 ).

[Example 2] In view of the results as described above, it is also meaningful to predict the prognosis of a patient with long stomach gastric cancer through the expression level of the VGLL1 gene or the HER2 gene. Through a method of checking whether it is possible, it may be possible to predict a more accurate survival rate.

Confirmation of the expression patterns of other genes in intestinal gastric cancer tissue

As confirmed in the above examples, the prognosis of a patient with long stomach gastric cancer can be predicted by measuring the expression levels of the VGLL1 and HER2 genes. At this time, if the expression levels of additional genes that are correlated with the survival rate are also measured, the prediction accuracy is measured. Since it can be increased, it was confirmed whether there are other genes showing specific expression levels in the intestinal gastric cancer tissue.

In order to find other genes that are specifically expressed at higher levels or specifically at lower levels, the Cox proportional hazard model is applied to all genes expressed in intestinal gastric cancer tissue. The hazard ratio was calculated. Based on this, it was possible to select the top 21 genes that could be expected to be related to the prognosis of patients with high risk gastrointestinal cancer (see FIG. 5).

Analysis of correlation between expression and survival rate of genes of [Example 4] in patients with long stomach gastric cancer

In order to analyze how the expression level of the 21 genes (refer to FIG. 5) additionally selected in [Example 4] actually correlates with the survival rate of patients with long-term gastric cancer, the VGLL1 gene and the HER2 gene are additionally derived. Based on the expression levels of the 21 genes, risk scores of patients with long stomach gastric cancer were calculated and classified into a high-risk group and a low-risk group (see A in FIG. 6 ).

Then, as a result of analyzing the overall survival rate between the high-risk group and the low-risk group, it was confirmed that the overall survival rate was significantly lower in the high-risk group compared to the low-risk group (see FIG. 6B ).

Through the above results, by measuring and analyzing the expression levels of the 21 genes in addition to the expression levels of the VGLL1 and HER2 genes, it is possible to increase accuracy in predicting the survival rate of patients with bowel cancer. Therefore, the prognosis of the patient with long stomach gastric cancer can be predicted by measuring the expression level of the genes through the composition and kit included in the present invention, and the accuracy of the prognosis prediction is compared with the measurement level of the gene expression level of the normal patient. High information can be provided.

In the above, the present invention has been described in detail only with respect to the described embodiments, but it is apparent to those skilled in the art that various modifications and variations are possible within the technical scope of the present invention, and it is natural that such modifications and modifications belong to the appended claims.

Claims (13)

VGLL1 (Vestigial Like Family Member 1) A composition for predicting prognosis of gastrointestinal cancer, comprising an agent that measures the expression level of a gene. The method according to claim 1,
A composition for predicting prognosis of gastrointestinal cancer, further comprising an agent that measures the expression level of the HER2 (Human Epidermal Growth Factor Receptor 2) gene. The method according to claim 2,
MMP9 ( Matrix Metallopeptidase 9 ), WNT7A (Wnt Family Member 7A), RTN3 (Reticulon 3), EDA (Ectodysplasin A), ROPN1B (Rhophilin Associated Tail Protein 1B), WIPF3 (WAS/WASL Interacting Protein Family Member 3), GCNT2 ( Glucosaminyl (N-Acetyl) Transferase 2), TM7SF3 (Transmembrane 7 Superfamily Member 3), CRABP2 (Cellular Retinoic Acid Binding Protein 2), KLK7 ( Kallikrein Related Peptidase 7), DEFB1 (Defensin Beta 1), RPS6 (Ribosomal Protein S6) , HNRPM (Heterogeneous Nuclear Ribonucleoprotein M), AURKAIP1 (Aurora Kinase A Interacting Protein 1), NASP (Nuclear Autoantigenic Sperm Protein), CPSF2 (Cleavage And Polyadenylation Specific Factor 2), GFI1 (Growth Factor Independent 1 Transcriptional Repressor), SNORD31 (Small Nucleolar RNA, C/D Box 31), RPSA (Ribosomal Protein SA), ELAVL1 (ELAV Like RNA Binding Protein 1), and further comprising an agent for measuring the expression level of one or more genes selected from the group consisting of LRRC48 , Composition for predicting long-term gastric cancer prognosis. The method according to any one of claims 1 to 3,
The composition for measuring the expression level of the gene is a composition for measuring the amount of mRNA of the gene, a formulation for measuring the amount of protein expressed from the gene, or both, and a composition for predicting prognosis of gastrointestinal cancer. The method according to claim 4,
The agent for measuring the amount of mRNA of the gene comprises a primer or probe specifically binding to the mRNA or cDNA of the gene, the composition for predicting long-term gastric cancer prognosis. The method according to claim 4,
The formulation for measuring the amount of protein expressed from the gene comprises an antibody or aptamer specific for the protein expressed from the gene, a composition for predicting prognosis of gastrointestinal cancer. A kit for predicting long-term gastric cancer prognosis, comprising the composition according to claim 1. Measuring the expression level of the VGLL1 gene from a biological sample of a patient with long stomach gastric cancer; And comparing the expression level of the measured gene with the expression level in a normal individual. Method for providing information for predicting the prognosis of a patient with long stomach gastric cancer. The method according to claim 8,
In the step of measuring the expression level of the gene, in addition to the expression level of the VGLL1 gene, further measuring the expression level of the HER2 gene, a method for providing information for predicting the prognosis of a patient with long stomach gastric cancer. The method according to claim 9,
In the step of measuring the expression level of the gene, in addition to the expression levels of the VGLL1 gene and the HER2 gene, MMP9, WNT7A, RTN3, EDA, ROPN1B, WIPF3, GCNT2, TM7SF3 CRABP2, KLK7, DEFB1, RPS6, HNRPM, AURKAIP1, A method of providing information for predicting the prognosis of a patient with bowel cancer, further measuring the expression level of one or more genes selected from the group consisting of NASP, CPSF2, GFI1, SNORD31, RPSA, ELAVL1, and LRRC48 . The method according to any one of claims 8 to 10,
The step of measuring the expression level of the gene is to measure the amount of mRNA of the gene, the amount of protein expressed from the gene, or both, to provide information for predicting the prognosis of a patient with long stomach gastric cancer Way. The method according to claim 11,
Measuring the amount of mRNA of the gene, using a primer or probe that specifically binds to the mRNA or cDNA of the gene, a method for providing information for predicting the prognosis of a patient with long stomach gastric cancer. The method according to claim 11,
Measuring the amount of protein expressed from the gene, using an antibody or aptamer specific for the protein expressed from the gene, a method for providing information for predicting the prognosis of a patient with long stomach gastric cancer.

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