KR102280503B1 - Composition for diagnosing cancer - Google Patents

Abstract

The present invention provides a composition, kit, and a composition that can predict the possibility of developing cancer early by measuring the expression level of ADAM8 or a gene encoding the same from a liquid biopsy of a subject of interest, and also predict the stage of cancer It relates to a method of providing information for prediction.

Description

Composition for diagnosing cancer

The present invention relates to a composition, a kit, and a method for providing information for prediction, which can predict the possibility of developing cancer at an early stage from a liquid biopsy of a subject of interest, and also predict the stage of cancer.

Gastric cancer is one of the most common cancers in the world in Korea, China, and Japan. The incidence rate is low in Western countries such as the United States and Europe, but in Korea, stomach cancer ranks first in cancer incidence and second in mortality after lung cancer. If we look at the classification of gastric cancer, 95% of all cancers are adenocarcinomas originating from the glandular cells of the gastric wall mucosa. Others include lymphomas originating in the lymphatic system and gastrointestinal stromal tumors originating in the interstitial tissue.

Among the current cancer treatment methods, there are three main treatments: surgical treatment, drug therapy, and radiation therapy. Drug therapy has less pain accompanying the treatment, and the recurrence of cancer after treatment is lower than surgery or radiation therapy. Since it is relatively small, expectations are being raised for this. Therefore, many anticancer drugs have been developed and used to respond to this, and most of these anticancer drugs exhibit anticancer effects by selectively killing actively dividing cells with the abnormal proliferation of cancer in mind. These anticancer drugs have a problem in that they cannot be used for a long period of time because they have serious side effects in that they also kill normal cells such as immune cells and hair follicles, which are cells that are actively dividing in the human body. In addition, methods such as lymph node dissection, endoscopic mucosal resection, and laparoscopic gastrectomy are used as methods for the treatment of gastric cancer. In endoscopic mucosal resection, physiological saline is administered around the gastric mucosa with cancer lesions for early gastric cancer within the mucosa. It is a method of excising the lesioned mucosa after injection to convexly inflate the lesion site. It has the advantage of avoiding the pain of gastrectomy with a simple endoscopic procedure. There are limits to what can be used.

Recently, worldwide, research is underway to discover therapeutic targets through functional studies of genes related to the generation and treatment of cancer and to use them for diagnosis and development of therapeutics. With the revitalization of genome research, human gene DNA chip or proteome analysis research is actively conducted and cancer-related genes are discovered in large numbers, a list of many genes and related databases have been established. Biological functions and cancer relevance have not yet been studied or are uncertain, so there is considerable difficulty in discovering genes that can effectively treat cancer as well as actual cancer relevance or diagnosis and use as target genes. Therefore, there is a need to discover new genes in addition to the cancer-related genes identified so far.

An object of the present invention is to provide a composition capable of diagnosing the possibility of developing cancer early and diagnosing the stage of cancer with high accuracy.

Another object of the present invention is to provide a kit capable of diagnosing the possibility of cancer at an early stage and the stage of cancer with high accuracy.

Another object of the present invention is to provide a method of providing information for diagnosing the possibility of cancer at an early stage and the stage of cancer with high accuracy.

Another object of the present invention is to provide a method for screening a drug for treating the cancer.

However, the technical task to be achieved by the present invention is not limited to the tasks mentioned above, and other tasks not mentioned will be clearly understood by those of ordinary skill in the art from the following description.

According to one embodiment of the present invention, it relates to a composition for diagnosing cancer, including an agent for measuring the expression level of A disintegrin and metalloproteinase domain-containing protein 8 (ADAM8) or a gene encoding the same.

In the present invention, the "ADAM8" is an enzyme encoded by the ADAM8 gene, and belongs to a membrane-anchored protein structurally related to snake venom disintegrins, and is an enzyme between the cell-cell and cell-matrix. It is involved in a number of biological processes. In addition, the ADAM8 has been known to be involved in cell adhesion during neurodegeneration. In the present invention, the ADAM8 may consist of the amino acid sequence represented by SEQ ID NO: 1, but is not limited thereto.

In the present invention, the expression level of the ADAM8 or a gene encoding it may be measured from a liquid biopsy of a subject of interest. In the present invention, the liquid biopsy may be blood, serum or plasma.

In general, in order to measure the expression level of a biomarker for the diagnosis of cancer, a part of the cancerous tissue has been isolated and then the expression level of the biomarker in the cancerous tissue has been measured. Therefore, in the early stage of cancer, it is difficult to recognize and it is rare to take tissue, so there is a disadvantage that an initial diagnosis is almost impossible, and there is also the inconvenience of isolating cells from the tissue.

However, in the present invention, by measuring the expression level of ADAM8 or a gene encoding it in a liquid biopsy of blood, serum or plasma isolated from a subject of interest, early diagnosis of cancer is possible, and the stage of cancer can also be predicted. there is.

The composition of the present invention may further include an agent for measuring the expression level of a cancer embryonic antigen (CEA) or a gene encoding the same, thereby increasing the accuracy of the cancer diagnosis.

In the present invention, the "carcinoembryonic antigen (CEA)" is a glycoprotein involved in cell adhesion, and is produced in the gastrointestinal tissue during fetal development, and its production stops before birth. In the present invention, the cancer embryo antigen may consist of the amino acid sequence represented by SEQ ID NO: 2, but is not limited thereto.

In the present invention, the agent for measuring the expression level of the ADAM8 or cancer embryo antigen is not particularly limited, but preferably an antibody, oligopeptide, ligand, PNA (peptide nucleic acid) that specifically binds to the ADAM8 or cancer embryo antigen protein. acid) and may include one or more selected from the group consisting of aptamers.

In the present invention, the term "measuring the expression level of a protein" refers to a process of confirming the presence and expression level of ADAM8, a diagnostic marker for cancer, or a protein of a cancer embryonic antigen in a biological sample for diagnosing cancer. The ADAM8 or cancer embryo antigen expression level measurement or comparative analysis method includes protein chip analysis, immunoassay, ligand binding assay, MALDI-TOF (Matrix Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry) analysis, SELDI-TOF ( Sulface Enhanced Laser Desorption/Ionization Time of Flight Mass Spectrometry (Sulface Enhanced Laser Desorption/Ionization Time of Flight Mass Spectrometry) analysis, radioimmunoassay, radioimmunodiffusion method, Oakteroni immunodiffusion method, rocket immunoelectrophoresis, tissue immunostaining, complement fixation assay, 2D electrophoresis analysis, liquid chromatography Graph-mass spectrometry (liquid chromatography-Mass Spectrometry, LC-MS), LC-MS/MS (liquid chromatography-Mass Spectrometry/ Mass Spectrometry), Western blotting and ELISA (enzyme linked immunosorbent assay), etc., but are limited thereto no.

In the present invention, the "antibody" refers to a substance that specifically binds to an antigen and causes an antigen-antibody reaction. For the purposes of the present invention, an antibody refers to an antibody that specifically binds to ADAM8 or a cancer embryonic antigen. Antibodies of the present invention include polyclonal antibodies, monoclonal antibodies and recombinant antibodies. The antibody can be readily prepared using techniques well known in the art. For example, the polyclonal antibody can be produced by a method well known in the art, which includes injecting the ADAM8 or an antigen of the cancer embryo antigen into an animal and collecting blood from the animal to obtain a serum containing the antibody. . Such polyclonal antibodies can be prepared from any animal such as goat, rabbit, sheep, monkey, horse, pig, cow, dog, and the like. Monoclonal antibodies can also be prepared using the hybridoma method well known in the art (see Kohler and Milstein (1976) European Journal of Immunology 6:511-519), or the phage antibody library technology (Clackson et al, Nature, 352). :624-628, 1991; Marks et al, J. Mol. Biol., 222:58, 1-597, 1991). The antibody prepared by the above method may be separated and purified using methods such as gel electrophoresis, dialysis, salt precipitation, ion exchange chromatography, and affinity chromatography. In addition, the antibodies of the present invention include 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 includes Fab, F(ab'), F(ab')2 and Fv.

In the present invention, "PNA (Peptide Nucleic Acid)" refers to an artificially synthesized, DNA or RNA-like polymer, and was first introduced by Professors Nielsen, Egholm, Berg and Buchardt of the University of Copenhagen, Denmark in 1991. Whereas DNA has a phosphate-ribose sugar backbone, PNA has a repeated N-(2-aminoethyl)-glycine backbone linked by peptide bonds, which greatly increases binding strength and stability to DNA or RNA, resulting in molecular biology , diagnostic assays and antisense therapy. PNA is described in Nielsen PE, Egholm M, Berg RH, Buchardt O (December 1991). "Sequence-selective recognition of DNA by strand displacement with a thymine-substituted polyamide". Science 254 (5037): 1497-1500.

In the present invention, the "aptamer" is an oligonucleic acid or a peptide molecule, and the general description of the aptamer is described in Bock LC et al., Nature 355(6360):5646(1992); Hoppe-Seyler F, Butz K "Peptide aptamers: powerful new tools for molecular medicine". J Mol Med. 78(8):42630(2000); Cohen BA, Colas P, Brent R. "An artificial cell-cycle inhibitor isolated from a combinatorial library". Proc Natl Acad Sci USA. 95(24): 142727 (1998).

In the diagnostic composition according to the present invention, the agent for measuring the expression level of the gene encoding the ADAM8 or cancer embryonic antigen consists of a primer, a probe and an antisense nucleotide that specifically binds to the ADAM8 or a gene encoding the cancer embryo antigen. It may include one or more selected from the group. Since information on ADAM8 and cancer embryonic antigens according to the present invention is known, those skilled in the art will be able to easily design primers, probes or antisense nucleotides that specifically bind to the gene encoding the protein based thereon.

In the present invention, the term "measuring the expression level of a gene" means measuring the amount of a gene by checking the presence and expression level of a gene encoding ADAM8 or cancer embryonic antigen in the liquid biopsy to diagnose cancer. . Analytical methods for this include reverse transcription polymerase reaction (RT-PCR), competitive reverse transcription polymerase reaction (Competitive RT-PCR), real-time reverse transcription polymerase reaction (Real-time RT-PCR), RNase protection assay (RPA; RNase protection) assay), Northern blotting, and a DNA chip, but is not limited thereto.

In the present invention, the "primer" is a fragment that recognizes a target gene sequence, and includes a pair of forward and reverse primers, but preferably, a primer pair that provides analysis results with specificity and sensitivity. High specificity can be conferred when the primer's nucleic acid sequence is a sequence that is inconsistent with the non-target sequence present in the sample, so that only the target gene sequence containing the complementary primer binding site is amplified and the primer does not cause non-specific amplification. .

In the present invention, the "probe" refers to a substance capable of specifically binding to a target substance to be detected in a sample, and refers to a substance capable of specifically confirming the presence of a target substance in a sample through the binding. The type of probe is not limited as a material commonly used in the art, but preferably PNA (peptide nucleic acid), LNA (locked nucleic acid), peptide, polypeptide, protein, RNA or DNA, and most preferably For example, it may be PNA. More specifically, the probe includes a biomaterial derived from or similar thereto or manufactured in vitro, for example, enzymes, proteins, antibodies, microorganisms, animal and plant cells and organs, neurons, DNA, and It may be RNA, DNA includes cDNA, genomic DNA, and oligonucleotides, RNA includes genomic RNA, mRNA, and oligonucleotides, and examples of proteins include antibodies, antigens, enzymes, peptides, and the like.

In the present invention, the "LNA (Locked nucleic acids)" means a nucleic acid analog including a 2'-O, 4'-C methylene bridge [J Weiler, J Hunziker and J Hall Gene Therapy (2006) 13, 496.502) ]. LNA nucleosides include common nucleic acid bases in DNA and RNA, and can form base pairs according to Watson-Crick base pairing rules. However, due to the 'locking' of the molecule due to the methylene bridge, the LNA does not form an ideal shape in the Watson-Crick bond. When LNAs are incorporated into DNA or RNA oligonucleotides, LNAs can pair with complementary nucleotide chains more rapidly, increasing the stability of the double helix. In the present invention, the "antisense" means that the antisense oligomer is hybridized with a target sequence in RNA by Watson-Crick base pairing, and typically mRNA and RNA in the target sequence: A sequence of nucleotide bases allowing the formation of an oligomeric heteroduplex. and oligomers having an inter-subunit backbone. An oligomer may have exact sequence complementarity or approximate complementarity to a target sequence.

In the present invention, the "diagnosis" refers to determining the susceptibility of a subject to a specific disease or disorder, determining whether the subject currently has a specific disease or disorder, or having a specific disease or disorder Determining a subject's prognosis (e.g., identifying a pre-metastatic or metastatic cancer state, staging the cancer, or determining the responsiveness of a cancer to treatment), or using therametrics (e.g., for therapeutic efficacy); monitoring the state of an object to provide information). For the purpose of the present invention, the diagnosis is to determine whether or not the disease occurs or the likelihood (risk) of the disease, the stage of the cancer, or the survival rate or treatment responsiveness of a cancer patient.

In the present invention, the "stage" refers to the extent to which cancer cells have spread and the stage of cancer progression, and the international classification according to the progress of cancer generally follows the TNM stage classification. Here, 'T(Tumor Size)' is a classification according to the size of the primary tumor, 'N(Lymph Node)' is a classification according to the degree of lymph node metastasis, and 'M(Metastasis)' is a classification according to whether it has metastasized to other organs. corresponds to Detailed classification for T, N, and M is shown in Table 1 below.

TNM Ordnance Justice size of primary tumor
(T stage)
Size of the primary tumor (T stage) T0 Tumor cells that show the appearance of a malignant tumor, but are confined to the mucous membrane or epithelium, and have not yet invaded the basement membrane T1 Limited lesions in the primary organ, the tumor is moving, and there is no invasion of adjacent and surrounding tissues T2 The size of the tumor is about 2-5 cm. T3 Tumor is larger than T2 but localized within an organ T4a Adhesion and infiltration with serosa tissue T4b Adhesion and infiltration with surrounding tissues Lymph node metastasis
(N stage)
Lymph node status (N stage) N0 No evidence of lymph node lesions N1 Invasion of a single palpable, mobile, limited lymph node (1 to 2 cm or larger, usually up to 3 cm in size) N2 Palpable, partially mobile or hard or hard lymph nodes, microscopically evidence of involvement, clinically entangled, contralateral or bilateral (3-5 cm) N3 Whoa?? It is fixed and passes through the capsule and is completely fixed to bones, large blood vessels, skin, nerves, etc., and has a size of 6 cm or more. Whether distant metastasis
(Army M)
Distant metastasis
(M stage) M0 No distant metastases M1 have distant metastases

In addition, by synthesizing the classification of TNM of the cancer, the stage of cancer can be classified into IA, IB, IIA, IIB, IIIA, IIIB, IIIC, IV as shown in Table 2 below, wherein the IA, IB, IIA and IIB The stage can be classified as an early stage of cancer.

ordnance T N M Weapon IA T1 N0 M0 Weapon IB T2 N0 M0 T1 N1 M0 Weapon IIA T3 N0 M0 T2 N1 M0 T1 N2 M0 Weapon IIB T4a N0 M0 T3 N1 M0 T2 N2 M0 T1 N3 M0 Weapon IIIA T4a N1 M0 T3 N2 M0 T2 N3 M0 Weapon IIIB T4b N0 M0 T4b N1 M0 T4a N2 M0 T3 N3 M0 Weapon IIIC T4b N2 M0 T4b N3 M0 T4a N3 M0 Weapon IV all T all N M1

ADAM8 from a liquid biopsy of a subject of interest, and optionally a cancer embryo antigen; Alternatively, by measuring the expression level of the gene encoding them, it is possible to diagnose whether or not the onset of early stage cancer or the possibility of the onset.

In addition, the present invention provides ADAM8, and optionally a cancer embryo antigen, from a liquid biopsy of a subject of interest; Alternatively, by measuring the expression level of the gene encoding them, it is possible to predict whether or not the cancer of N-stage will develop or the possibility of developing cancer, or which stage the cancer belongs to in the N-stage.

In the present invention, the term "cancer" as a disease to be diagnosed refers to or refers to a physiological condition characterized by uncontrolled cell growth typically in mammals. Cancers to be diagnosed in the present invention include gastric cancer, ovarian cancer, colorectal cancer, pancreatic cancer, liver cancer, breast cancer, cervical cancer, thyroid cancer, parathyroid cancer, lung cancer, non-small cell lung cancer, prostate cancer, gallbladder cancer, biliary tract cancer, and non-Hodgkin's lymphoma. , Hodgkin's lymphoma, blood cancer, bladder cancer, kidney cancer, melanoma, colon cancer, bone cancer, skin cancer, head cancer, uterine cancer, rectal cancer, brain tumor, perianal cancer, fallopian tube carcinoma, endometrial carcinoma, vaginal cancer, vulvar carcinoma, esophageal cancer, small intestine Cancer, endocrine adenocarcinoma, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, ureter cancer, renal cell carcinoma, renal pelvic carcinoma, CNS central nervoussystem tumor, primary CNS lymphoma, spinal cord tumor, brainstem glioma or pituitary gland It may be an adenoma, but preferably gastric cancer.

According to another embodiment of the present invention, it relates to a kit for diagnosis of cancer comprising the composition for diagnosis of cancer of the present invention.

In the present invention, by diagnosing the onset or possibility of early stage cancer using the diagnostic kit, early diagnosis of cancer is possible, and the stage of cancer, preferably N-stage, can be predicted. .

In the present invention, as the disease to be diagnosed, cancer is gastric cancer, ovarian cancer, colorectal cancer, pancreatic cancer, liver cancer, breast cancer, cervical cancer, thyroid cancer, parathyroid cancer, lung cancer, non-small cell lung cancer, prostate cancer, gallbladder cancer, biliary tract cancer, Non-Hodgkin's lymphoma, Hodgkin's lymphoma, blood cancer, bladder cancer, kidney cancer, melanoma, colon cancer, bone cancer, skin cancer, head cancer, uterine cancer, rectal cancer, brain tumor, perianal cancer, fallopian tube carcinoma, endometrial carcinoma, vaginal cancer, vulvar carcinoma, Esophageal cancer, small intestine cancer, endocrine adenocarcinoma, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, ureter cancer, renal cell carcinoma, renal pelvic carcinoma, CNS central nervoussystem tumor, primary CNS lymphoma, spinal cord tumor, brainstem nerve glioma or pituitary adenoma, but preferably gastric cancer.

In the present invention, the kit may be an RT-PCR kit, a DNA chip kit, an ELISA kit, a protein chip kit, a rapid kit, or a multiple reaction monitoring (MRM) kit, but is not limited thereto.

The cancer diagnostic kit of the present invention may further include one or more other component compositions, solutions or devices suitable for the analysis method.

For example, in the present invention, the diagnostic kit for cancer may further include essential elements necessary for performing a reverse transcription polymerase reaction. The reverse transcription polymerase reaction kit includes a pair of primers specific for a gene encoding a marker protein. The primer is a nucleotide having a sequence specific to the nucleic acid sequence of the gene, and may have a length of about 7 bp to 50 bp, more preferably about 10 bp to 30 bp. It may also include primers specific for the nucleic acid sequence of the control gene. Other reverse transcription polymerase reaction kits include test tubes or other suitable containers, reaction buffers (with varying pH and magnesium concentrations), deoxynucleotides (dNTPs), enzymes such as Taq-polymerase and reverse transcriptase, DNase, RNase inhibitor DEPC -Water (DEPC-water), sterile water, etc. may be included.

In addition, the diagnostic kit of the present invention may include essential elements necessary for performing a DNA chip. The DNA chip kit may include a substrate to which cDNA or oligonucleotide corresponding to a gene or fragment thereof is attached, and reagents, agents, enzymes, etc. for preparing a fluorescently-labeled probe. The substrate may also contain cDNA or oligonucleotides corresponding to control genes or fragments thereof.

In addition, the diagnostic kit of the present invention may include essential elements necessary for performing ELISA. The ELISA kit contains an antibody specific for this protein. Antibodies are antibodies with high specificity and affinity for a marker protein and little cross-reactivity with other proteins, and are monoclonal antibodies, polyclonal antibodies, or recombinant antibodies. The ELISA kit may also include an antibody specific for a control protein. Other ELISA kits include reagents capable of detecting bound antibody, such as labeled secondary antibodies, chromophores, enzymes (eg, conjugated with an antibody) and substrates thereof or capable of binding the antibody. other materials and the like.

According to another embodiment of the present invention, there is provided a method for diagnosing cancer or providing information on the stage of cancer, comprising measuring the expression level of ADAM8 or a gene encoding the same in a biological sample isolated from a subject of interest will be.

In the present invention, the "target individual" refers to an individual who is uncertain whether or not the onset of the cancer has a high probability of developing it.

In the present invention, the "biological sample" is a liquid biopsy obtained from or derived from an individual, and in the present invention, the liquid biopsy may be blood, serum or plasma.

The present invention may include measuring the expression level of ADAM8 or a gene encoding the same in the biological sample isolated as described above.

In addition, in the present invention, by further comprising the step of measuring the expression level of a cancer embryonic antigen (CEA) or a gene encoding the same with respect to the biological sample isolated as described above, the accuracy of the cancer diagnosis can be increased.

In the present invention, the agent for measuring the expression level of the ADAM8 or cancer embryo antigen is not particularly limited, but preferably an antibody, oligopeptide, ligand, PNA (peptide nucleic acid) that specifically binds to the ADAM8 or cancer embryo antigen protein. acid) and may include one or more selected from the group consisting of aptamers.

In the present invention, as a method for measuring or comparing the expression level of ADAM8 or cancer embryonic antigen, protein chip analysis, immunoassay, ligand binding assay, MALDI-TOF (Matrix Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry) analysis, SELDI-TOF (Sulface Enhanced Laser Desorption/Ionization Time of Flight Mass Spectrometry) analysis, radioimmunoassay, radioimmunodiffusion method, Oukteroni immunodiffusion method, rocket immunoelectrophoresis, tissue immunostaining, complement fixation assay, two-dimensional electrophoresis analysis, liquid chromatography-Mass Spectrometry (LC-MS), LC-MS/MS (liquid chromatography-Mass Spectrometry/ Mass Spectrometry), Western blotting and ELISA (enzyme linked immunosorbent assay), etc. It is not limited thereto.

In the present invention, the agent for measuring the expression level of the gene encoding the ADAM8 or cancer embryo antigen is one selected from the group consisting of primers, probes and antisense nucleotides that specifically bind to the ADAM8 or cancer embryo antigen encoding gene. may include more than one.

In the present invention, as a process of confirming the presence and expression level of the gene encoding the ADAM8 or cancer embryonic antigen, the analysis method for measuring the amount of the gene includes a reverse transcription polymerase reaction (RT-PCR), a competitive reverse transcription polymerase reaction ( Competitive RT-PCR), real-time RT-PCR, RNase protection assay (RPA), Northern blotting, DNA chip, etc., but are not limited thereto.

ADAM8, and optionally cancer embryonic antigen, measured against a biological sample of a subject of interest in the present invention; Alternatively, it may include predicting the likelihood of developing cancer or the stage of cancer by comparing the expression level of the gene encoding them with a level measured in a normal control group.

In the present invention, it is possible to predict the possibility or the onset of cancer in an early stage, so that early diagnosis of cancer is possible.

Also, in the present invention, the stage of the cancer may be N-stage regarding whether or not metastasis to the lymph nodes is present.

ADAM8, and optionally cancer embryonic antigen, measured against a biological sample of a subject of interest in the present invention; Or, when the expression level of the gene encoding them is increased compared to the normal control, it may include predicting that the likelihood of developing the cancer is high.

In addition, in the present invention, when the expression level of ADAM8 measured in the biological sample of the subject of interest is 20 ng/ml or more, 25 ng/ml or more, or 30 ng/ml or more, preferably 25 ng/ml or more, the cancer is It can be predicted that there is a high probability of

In the present invention, when the expression level of the cancer embryonic antigen measured in the biological sample of the subject of interest is 3 ng/ml or more or 5 ng/ml or more, the possibility of developing the cancer can be predicted to be high.

In addition, in the present invention, ADAM8 measured in the biological sample of the subject of interest, and optionally a cancer embryo antigen; Or, when the expression level of the gene encoding them is increased compared to the normal control group, predicting that the cancer progression is N-stage, or that the cancer has a high possibility of lymph node metastasis. can

In the present invention, the cancer is gastric cancer, ovarian cancer, colorectal cancer, pancreatic cancer, liver cancer, breast cancer, cervical cancer, thyroid cancer, parathyroid cancer, lung cancer, non-small cell lung cancer, prostate cancer, gallbladder cancer, biliary tract cancer, non-Hodgkin's lymphoma, Hodgkin's lymphoma , blood cancer, bladder cancer, kidney cancer, melanoma, colon cancer, bone cancer, skin cancer, head cancer, uterine cancer, rectal cancer, brain tumor, perianal cancer, fallopian tube carcinoma, endometrial carcinoma, vaginal cancer, vulvar carcinoma, esophageal cancer, small intestine cancer, endocrine adenocarcinoma , adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, ureter cancer, renal cell carcinoma, renal pelvic carcinoma, CNS central nervoussystem tumor, primary CNS lymphoma, spinal cord tumor, brainstem glioma or pituitary adenoma , preferably gastric cancer.

According to another embodiment of the present invention, measuring the expression level of ADAM8 or a gene encoding the same in the isolated biological sample;

contacting a test substance with the biological sample; and

It relates to a method of screening a drug for treating cancer, comprising measuring the expression level of the ADAM8 or the gene in the biological sample after contacting the test substance.

In the present invention, by further comprising the step of measuring the expression level of a cancer embryonic antigen (CEA) or a gene encoding the same with respect to the biological sample isolated as described above, the accuracy in screening a drug for treating the cancer can increase

In the present invention, the test substance includes any substance, molecule, element, compound, entity, or a combination thereof. Examples include, but are not limited to, proteins, polypeptides, small organic molecules, polysaccharides, polynucleotides, and the like. It may also be a natural product, a synthetic compound, or a combination of two or more substances.

In the present invention, the ADAM8, and optionally a cancer embryonic antigen (CEA) in the biological sample after contact with the test substance; Alternatively, when the expression level of the gene encoding the same is reduced compared to before contact with the test substance, the method may further include determining the test substance as a therapeutic agent for cancer.

In addition, in the present invention, the ADAM8, and optionally cancer embryonic antigen (CEA) in the biological sample after contact with the test substance; Alternatively, when the expression level of the gene encoding the same is reduced compared to before contact with the test substance, the method may further include determining the test substance as a cancer lymph node metastasis inhibitor.

In the present invention, the subject, the biological sample and the definition of cancer, ADAM8 or cancer embryo antigen; Alternatively, with respect to a method of measuring the expression level of a gene encoding them, the detailed description thereof will be omitted below in order to avoid excessive complexity of the specification because it overlaps with that described in the information providing method for diagnosis of cancer of the present invention.

According to another embodiment of the present invention, the method comprising: treating the isolated biological sample with a candidate substance expected to induce cancer; and

It relates to a method for screening a cancer-inducing drug, comprising measuring the expression level of ADAM8 or a gene encoding the same in a biological sample treated with the candidate substance.

In the present invention, by further comprising the step of measuring the expression level of a cancer embryonic antigen (CEA) or a gene encoding the same with respect to the biological sample isolated as described above, the accuracy of screening the cancer-inducing drug is improved. can be raised

In the present invention, the candidate material includes any substance, molecule, element, compound, entity, or a combination thereof. Examples include, but are not limited to, proteins, polypeptides, small organic molecules, polysaccharides, polynucleotides, and the like. It may also be a natural product, a synthetic compound, or a combination of two or more substances.

In the present invention, the ADAM8, and optionally a cancer embryonic antigen (CEA) in the biological sample after treatment with the candidate substance; Alternatively, when the expression level of the gene encoding the same is increased compared to before the treatment of the candidate material, the method may further include determining the candidate material as a cancer inducer.

In addition, in the present invention, the ADAM8, and optionally a cancer embryonic antigen (CEA) in the biological sample after treatment with the candidate substance; Alternatively, when the expression level of the gene encoding the same is increased compared to before the treatment of the candidate substance, the method may further include determining the candidate substance as a cancer lymph node metastasis inducer.

In the present invention, the subject, the biological sample and the definition of cancer, ADAM8 or cancer embryo antigen; Alternatively, with respect to a method of measuring the expression level of a gene encoding them, the detailed description thereof will be omitted below in order to avoid excessive complexity of the specification because it overlaps with that described in the information providing method for diagnosis of cancer of the present invention.

In the present invention, by measuring the expression level of ADAM8 or a gene encoding it with respect to a liquid biopsy of a subject of interest, cancer can be diagnosed early, the N-stage of cancer can be predicted, and a tissue related to a disease can be conventionally taken. Compared to the method of measuring the expression level of a biomarker, it has the advantage of being non-invasive, simple, rapid, but very accurate.

1 shows AUC and ROC curves of ADAM8 and CEA in blood for gastric cancer prediction in independent verification data according to an embodiment of the present invention.

Hereinafter, the present invention will be described in detail by the following examples. However, the following examples are only illustrative of the present invention, and the content of the present invention is not limited by the following examples.

Example

test subject

The test subjects were classified into 5 groups including gastritis-dysplasia-carcinoma. The normal control group includes individuals with normal gastric mucus or simple gastritis, the high-risk group includes individuals with low-grade or high-grade dysplasia, and the early gastric cancer (EGC) group includes individuals with gastric cancer limited to the submucosal layer without metastasis Including metastases, the advanced gastric cancer (AGC) group includes individuals with gastric cancer beyond the muscle layer without metastasis, and the AGC group with metastases includes individuals with gastric cancer beyond the muscle layer, including metastases.

A total of 80 subjects were selected as the initial training dataset. Among them, 20 subjects in the normal control group, 10 subjects in the dysplastic group, 20 subjects in the EGC group, 20 subjects in the AGC group without metastases, and 20 subjects in the AGC group with metastases. Ten people were registered.

A total of 241 subjects were selected as an independent validation dataset. Among them, 70 subjects in the normal control group, 24 subjects in the dysplastic group, 70 subjects in the EGC group, 50 subjects in the AGC group without metastases, and 50 subjects in the AGC group with metastases. 27 people were enrolled.

In order to identify new markers for gastric cancer diagnosis and staging of gastric cancer that can improve diagnostic accuracy, gastrointestinal endoscopy was performed for all patients who visited Yonsei University Health System for gastric symptoms or routine examination. endoscopy) (Types XQ-260, Olympus, Tokyo, Japan) was performed, and blood samples were obtained before further treatment was performed. For all gastric cancer patient groups, chest X-ray, abdominal-pelvic spiral CT, and whole body positron emission tomography (PET) were performed for TNM staging. Stomach cancer was classified according to the TMN stage of the 7th International Union Against Cancer. Histopathologic differentiation was diagnosed using Lauren classification. Patients suffering from chronic diseases such as liver cirrhosis, chronic kidney disease, and diabetes were excluded from this study. Patients with other carcinomas or other gastric tumors such as gastrointestinal stromal tumors, mucosa-associated lymphoid tissue lymphomas and neuroendocrine tumors were also excluded from this study. In addition, patients with a history of treatment for gastric cancer or premalignant lesions were also excluded from this study.

ADAM8 in blood; CEA; interleukin-23 (IL-23); stromal cell derived factor 1α (SDF-1α)/CXC chemokine ligand 12 (CXCL12); Determination of expression levels of interleukin-8 (IL-8) and soluble CD40 ligand (sCD40L)

The expression level of ADAM8 in blood was measured using a commercially available ELISA kit (SEA620Hu, CLOUD-CLONE, Katy, TX, USA). Briefly, 100 μl each of a blood sample and a standard solution were added to each well of a pre-coated 96-well plate, and reacted at 37° C. for 1 hour. Thereafter, 100 μl of detection reagent A was treated and reacted at 37° C. for 1 hour. After each well was washed 3 times, 100 μl of Reagent B was treated and reacted at 37° C. for 30 minutes. After washing 5 times, 90 μl of the substrate solution was added and reacted at 37° C. for 10-20 minutes. Thereafter, the expression level of ADAM8 in blood was measured at 450 nm using a VERSA max microplate reader (Molecular Devices Co., Sunnyvale, Calif., USA).

CEA expression levels in blood were measured using the Beckman Access CEA assay (Beckman Coulter Inc. Chaska, USA). Blood levels of IL-23, SDF-1 alpha, IL-8 and sCD40L were measured using a commercially available MILLIPLEX MAP Human Cytokine/Chemokine kit (Millipore, Billerica, MA, USA) using a chemiluminescent immunoassay. was used. Specifically, the filter plate was pre-immersed in 200 μl of assay buffer at room temperature for 10 minutes, and then the assay buffer was vacuum removed. 25 μl of standard or control solution was added to the wells and 25 μl of assay buffer was added to the sample wells, but not to the background wells. 25 μl of the appropriate substrate solution was added to the background wells, standard wells and control wells, and 25 μl of the sample was added. After mixing, 25 μl of beads was added and the plate was shaken and incubated overnight at 4°C. After incubation, the fluid was removed and the plates were washed twice. After addition of the detection antibody (25 μl), the plate was incubated for 1 hour at room temperature. Streptavidin-phycoerythrin (25 μl) was added to each well containing 25 μl of the detection antibody, and then incubated for 1 hour at room temperature with shaking. After the fluid was removed, the plate was washed and 150 μl of the sheath fluid was added. After re-suspending for 5 minutes, Luminex 100?? After reading the intermediate fluorescence intensity using IS, it was analyzed using a logistic curve-fitting method to determine the chemokine concentration.

statistical analysis

All measured values were expressed as mean values including standard deviation values of 25 to 75%, and each group mean value was one-way ANOVA test method using multiple comparisons of the post-hoc Bonferroni method. compared with An independent sample t-test was used to compare means in cancer and non-cancer conditions. And in order to confirm the correlation between the serum concentration measured through the experiment and clinical pathological parameters, Pearson's correlation, coefficient, g _p ) and Spearman's correlation coefficient (Spearman's correlation, coefficient, g _s ). The initial gastric cancer size was < 3 cm to analyze the correlation with the level of ADAM8 expression in blood; They were divided into three groups: 3-5 cm and >5 cm. After forming the ROC (receiver operating characteristic) curve, AUC (area under the curves) was calculated to compare the diagnostic accuracy of each value for predicting the presence of gastric cancer. Logistic regression analysis was performed to obtain optimal sensitivity/specificity between single markers or multiple markers in predicting the presence of gastric cancer. Each marker was included as a linear term.

Initial training data- Comparison of ADAM8 and CEA expression levels in blood in 5 groups

A total of 80 individuals were registered for initial training data. The expression levels of ADAM8 and CEA in the blood were measured during gastric cancer carcinogenesis, and the expression levels were compared in a total of five groups: a normal control group, a dysplastic group, an EGC group, an AGC group without metastases, and an AGC group with metastases. As a result, as shown in Table 3 below, the average value of the level of ADAM8 in the blood in the five groups was measured completely differently (ANOVA, p < 0.001), and the expression level increased as the cancerous process progressed: normal/gastritis The expression level of ADAM8 in blood in the group was 1.7±2.6 ng/mL, 15.3±13.3 ng/mL in the dysplastic group, 53.9±36.9 ng/mL in the EGC group, 82.8±52.6 ng/mL in the AGC group without metastases, primary The AGC group with metastases measured 53.6±26.3 ng/mL. It was confirmed that the level of ADAM8 expression in the blood was significantly increased in the early stage of gastric cancer. As shown in Table 3, the expression level of ADAM8 in the blood in the early gastric cancer patient group and EGC group was high-risk, post-hoc Bonferroni, p < 0.001) or normal control group (control, post-hoc Bonferroni, p). < 0.001), it was confirmed that it was significantly higher. However, the expression level of ADAM8 in the blood was not significantly different in each group of gastric cancer patients, and the expression level was slightly increased in the advanced gastric cancer patient group (AGC) without metastasis compared to the early gastric cancer patient group (EGC) (post-hoc Bonferroni between EGC) and AGC, p = 0.086), there was no statistical significance in the advanced gastric cancer patient group with primary metastasis, but the expression level was rather decreased. In addition, as shown in Table 4, the average value of ADAM8 expression levels in the blood was compared between the cancer patient group and the non-cancer patient group, and as a result, the cancer patient group showed 65.4±44.0 ng/mL and the non-cancer patient group (6.2±10.1 ng). /mL) showed a significantly higher value (t-test, p < 0.001).

On the other hand, as shown in Table 3, the expression level of CEA in the blood did not differ significantly between the groups except for the AGC group with metastases, and only significantly increased in the AGC group with metastases (ANOVA, p = 0.013). . When the mean expression level of CEA in the blood was compared between the cancer patient group and the non-cancer patient group, there was no significant difference in the expression level between the two groups (p = 0.235, t-test).

group
(N) Normal/Gastritis
(20) dysplasia
(10) EGC
(20) AGC
(20) transition
(10) p -value Serum ADAM8
(ng/mL) 1.7 ± 2.6 15.3 ± 13.3 53.9 ± 36.9 82.8 ± 52.6 53.6 ± 26.3 <0.001 Serum CEA (ng/mL) 1.9 ± 1.1 3.7 ± 1.9 2.0 ± 1.2 3.6 ± 6.7 9.8 ± 14.1 0.013

group
(N) Non-cancer patient population
(30) cancer patient group
(50) p -value Serum ADAM8 (ng/mL) 6.2 ± 10.1 65.4 ± 44.0 <0.001 Serum CEA (ng/mL) 2.5 ± 1.6 4.2 ± 7.9 0.235

Independent verification data - Comparison of ADAM8 and CEA expression levels in blood according to cancer progression of gastric cancer

In order to confirm the reproducibility of the results of the previous initial training data, independent verification data were used. A total of 241 individuals were registered in the verification data. Similar to the training data, the mean value of ADAM8 expression level in blood increased with cancer progression. According to Table 5, the expression level of ADAM8 in the blood was 6.4±8.7 ng/mL in the normal/gastritis group, 23.7±16.0 ng/mL in the dysplastic group, 54.5±42.7 ng/mL in the EGC group, and AGC group without metastasis. It was 67.9±55.1 ng/mL and 55.0±29.7 ng/mL in the AGC group with primary metastasis (ANOVA, p < 0.001). In the independent validation data, the expression level of ADAM8 in the blood was significantly increased in the early gastric cancer stage, and the expression level of ADAM8 in the blood was slightly increased in the advanced gastric cancer stage without metastasis compared to the early gastric cancer stage, but there was a significant difference between the cancer patient groups. there was no In addition, as shown in Table 6, as a result of comparing the average value of the expression level of ADAM8 in the blood in the cancer patient group and the non-cancer patient group, the same as the initial training data, the expression level of ADAM8 in the blood in the cancer patient group ( 59.1±45.4 ng/mL) showed a significantly higher value than the value of the expression level of ADAM8 in the blood (10.8±13.3 ng/mL) measured in the non-cancer patient group (t-test, p<0.001).

On the other hand, the level of CEA expression in blood also increased significantly only in the AGC group with metastasis, similar to the initial training data, and there was no significant difference between the cancer patient group and the non-cancer patient group.

group
(N) Normal/Gastritis
(70) dysplasia
(24) EGC
(70) AGC
(50) transition
(27) p -value Serum ADAM8 (ng/mL) 6.4 ± 8.7 ^?? 23.7 ± 16.0 54.5 ± 42.7 67.9 ± 55.1 55.0 ± 29.7 <0.001 Serum CEA (ng/mL) 1.9 ± 1.0 3.5 ± 4.3 2.1 ± 1.4 2.5 ± 2.8 6.7 ± 4.3 <0.001

group
(N) Non-cancer patient population
(94) cancer patient group
(148) p -value Serum ADAM8 (ng/mL) 10.8 ± 13.3 59.1 ± 45.4 <0.001 Serum CEA (ng/mL) 2.3 ± 2.4 3.0 ± 3.1 0.053

Identification of the correlation between the expression level of ADAM8 in blood and clinical pathological characteristics through validation data

As shown in Table 7 below, the level of ADAM8 expression in the blood was not affected by the subject's age (Pearson's correlation; γp = -0.064, p = 0.319) and gender (Spearman's correlation; γs = -0.095, p = 0.143). . Histopathologically, the expression level of ADAM8 in serum was determined by the histologic differentiation of gastric cancer (γs = 0.139, p = 0.193), the location of the early gastric cancer (γs = 0.058, p = 0.397), and the size of the early gastric cancer (γs = 0.121). , p = 0.249) and there was no significant correlation. In addition, the expression level of ADAM8 in the serum significantly correlated with the depth of cancer cell invasion (T-stage, γs = 0.197, p = 0.085) and primary metastasis (M-stage, γs = 0.140, p = 0.193). , but showed a high correlation with N-stage (γs = 0.320, p = 0.011)

Clinicopathological Characteristics Serum ADAM8 g _s ( p- value) age (years) -0.064 (0.319) Gender Male Female) -0.095 (0.143) Tissue (well differentiated: moderately differentiated: poorly differentiated: ring-shaped gastric cancer) 0.139 (0.193) Tumor location (lower:middle:upper) 0.058 (0.397) Tumor size (<3 cm; 3-5 cm and >5 cm) 0.121 (0.249) T-Stage (T1a:T1b:T2:T3:T4) 0.197 (0.085) N-Army (N0:N1:N2:N3) 0.320 (0.011) Primary Transition (M0:M1) 0.140 (0.193)

Diagnostic application of blood ADAM8, CEA and combinations thereof to predict the presence of gastric cancer through validation data

As a result of confirming the ROC curves and the measured AUC values of ADAM8 and CEA in blood for prediction of gastric cancer in the validation data, as shown in FIG. 1, the AUC of ADAM8 in blood was 0.90 (95% CI, 0.86-0.33), whereas The AUC of blood CEA was only 0.56 (95% CI, 0.49-0.63), suggesting that ADAM8 in blood has superior diagnostic accuracy compared to blood CEA in predicting the presence of gastric cancer. In addition, as shown in Table 8, in predicting the presence of gastric cancer by logistic regression analysis, the sensitivity and specificity of ADAM8 in blood were measured to be 73.7% and 86.2% (cut-off value: 30 ng/mL), respectively. , the sensitivity and specificity of CEA in blood were only 23.1% and 91.4%, respectively (cut-off value: 4 ng/mL).

Since the increasing patterns of ADAM8 and CEA in the blood in cancer progression are significantly different, it was predicted that there would be a synergistic effect when the two markers were used in combination. As a result, as shown in Table 8 below, it was confirmed that the accuracy of diagnosis was significantly increased when ADAM8 in blood and CEA in blood were used as single markers, respectively, when they were used in combination (sensitivity: 81.8%, specificity) : 84.0%).

marker cut-off value sensitivity specificity ADAM8 30 ng/mL 73.7% 86.2% CEA 5 ng/mL 23.1% 91.4% ADAM8 + CEA 25, 5 ng/mL 81.8% 84.0%

Identification of the correlation between the expression level of ADAM8 in blood and cancer-related cytokines through validation data

In order to confirm the role of ADAM8 in the blood in the progression of gastric cancer, the correlation between the expression level of ADAM8 in the blood and gastric cancer-related pre-inflammatory or pro-angiogenic cytokines in the validation data The relationship was analyzed. As a result, as shown in Table 9 below, ADAM8 in blood is a pro-inflammatory cytokine, IL-23 in blood (γp = 0.235, p = 0.036) and SDF-1α/CXCL12 in blood (γp = -0.233, p = 0.037) ), while not significantly correlated with the angiogenic cytokines, IL-8 in blood (γp = 0.113, p = 0.313) and sCD40L in blood (γp = 0.043, p = 0.702). could From this, it was found that ADAM8 in the blood was associated with cancer-related inflammation or local infiltration in the early stages of gastric cancer, but not with angiogenesis for primary metastasis.

IL-23
g _p ( p- value) SDF-1a
g _p ( p- value) IL-8
g _p ( p- value) sCD40L
g _p ( p- value) ADAM8 0.235 ( 0.036) -0.233 (0.037) 0.113 (0.313) 0.043 (0.702)

<110> Industry-Academic Cooperation Foundation, Yonsei University <120> Composition for diagnosis cancer <130> PDPB193257 <160> 2 <170> KoPatentIn 3.0 <210> 1 <211> 824 <212> PRT <213> Homo sapiens <400> 1 Met Arg Gly Leu Gly Leu Trp Leu Leu Gly Ala Met Met Leu Pro Ala 1 5 10 15 Ile Ala Pro Ser Arg Pro Trp Ala Leu Met Glu Gln Tyr Glu Val Val 20 25 30 Leu Pro Arg Arg Leu Pro Gly Pro Arg Val Arg Arg Ala Leu Pro Ser 35 40 45 His Leu Gly Leu His Pro Glu Arg Val Ser Tyr Val Leu Gly Ala Thr 50 55 60 Gly His Asn Phe Thr Leu His Leu Arg Lys Asn Arg Asp Leu Leu Gly 65 70 75 80 Ser Gly Tyr Thr Glu Thr Tyr Thr Ala Ala Asn Gly Ser Glu Val Thr 85 90 95 Glu Gln Pro Arg Gly Gln Asp His Cys Phe Tyr Gln Gly His Val Glu 100 105 110 Gly Tyr Pro Asp Ser Ala Ala Ser Leu Ser Thr Cys Ala Gly Leu Arg 115 120 125 Gly Phe Phe Gln Val Gly Ser Asp Leu His Leu Ile Glu Pro Leu Asp 130 135 140 Glu Gly Gly Glu Gly Gly Arg His Ala Val Tyr Gln Ala Glu His Leu 145 150 155 160 Leu Gln Thr Ala Gly Thr Cys Gly Val Ser Asp Asp Ser Leu Gly Ser 165 170 175 Leu Leu Gly Pro Arg Thr Ala Ala Val Phe Arg Pro Arg Pro Gly Asp 180 185 190 Ser Leu Pro Ser Arg Glu Thr Arg Tyr Val Glu Leu Tyr Val Val Val 195 200 205 Asp Asn Ala Glu Phe Gln Met Leu Gly Ser Glu Ala Ala Val Arg His 210 215 220 Arg Val Leu Glu Val Val Asn His Val Asp Lys Leu Tyr Gln Lys Leu 225 230 235 240 Asn Phe Arg Val Val Leu Val Gly Leu Glu Ile Trp Asn Ser Gln Asp 245 250 255 Arg Phe His Val Ser Pro Asp Pro Ser Val Thr Leu Glu Asn Leu Leu 260 265 270 Thr Trp Gln Ala Arg Gln Arg Thr Arg Arg His Leu His Asp Asn Val 275 280 285 Gln Leu Ile Thr Gly Val Asp Phe Thr Gly Thr Thr Val Gly Phe Ala 290 295 300 Arg Val Ser Ala Met Cys Ser His Ser Ser Gly Ala Val Asn Gln Asp 305 310 315 320 His Ser Lys Asn Pro Val Gly Val Ala Cys Thr Met Ala His Glu Met 325 330 335 Gly His Asn Leu Gly Met Asp His Asp Glu Asn Val Gln Gly Cys Arg 340 345 350 Cys Gln Glu Arg Phe Glu Ala Gly Arg Cys Ile Met Ala Gly Ser Ile 355 360 365 Gly Ser Ser Phe Pro Arg Met Phe Ser Asp Cys Ser Gln Ala Tyr Leu 370 375 380 Glu Ser Phe Leu Glu Arg Pro Gln Ser Val Cys Leu Ala Asn Ala Pro 385 390 395 400 Asp Leu Ser His Leu Val Gly Gly Pro Val Cys Gly Asn Leu Phe Val 405 410 415 Glu Arg Gly Glu Gln Cys Asp Cys Gly Pro Glu Asp Cys Arg Asn 420 425 430 Arg Cys Cys Asn Ser Thr Thr Cys Gln Leu Ala Glu Gly Ala Gln Cys 435 440 445 Ala His Gly Thr Cys Cys Gln Glu Cys Lys Val Lys Pro Ala Gly Glu 450 455 460 Leu Cys Arg Pro Lys Lys Asp Met Cys Asp Leu Glu Glu Phe Cys Asp 465 470 475 480 Gly Arg His Pro Glu Cys Pro Glu Asp Ala Phe Gln Glu Asn Gly Thr 485 490 495 Pro Cys Ser Gly Gly Tyr Cys Tyr Asn Gly Ala Cys Pro Thr Leu Ala 500 505 510 Gln Gln Cys Gln Ala Phe Trp Gly Pro Gly Gly Gln Ala Ala Glu Glu 515 520 525 Ser Cys Phe Ser Tyr Asp Ile Leu Pro Gly Cys Lys Ala Ser Arg Tyr 530 535 540 Arg Ala Asp Met Cys Gly Val Leu Gln Cys Lys Gly Gly Gln Gln Pro 545 550 555 560 Leu Gly Arg Ala Ile Cys Ile Val Asp Val Cys His Ala Leu Thr Thr 565 570 575 Glu Asp Gly Thr Ala Tyr Glu Pro Val Pro Glu Gly Thr Arg Cys Gly 580 585 590 Pro Glu Lys Val Cys Trp Lys Gly Arg Cys Gln Asp Leu His Val Tyr 595 600 605 Arg Ser Ser Asn Cys Ser Ala Gln Cys His Asn His Gly Val Cys Asn 610 615 620 His Lys Gln Glu Cys His Cys His Ala Gly Trp Ala Pro Pro His Cys 625 630 635 640 Ala Lys Leu Leu Thr Glu Val His Ala Ala Ser Gly Ser Leu Pro Val 645 650 655 Leu Val Val Val Val Leu Val Leu Leu Ala Val Val Leu Val Thr Leu 660 665 670 Ala Gly Ile Ile Val Tyr Arg Lys Ala Arg Ser Arg Ile Leu Ser Arg 675 680 685 Asn Val Ala Pro Lys Thr Thr Met Gly Arg Ser Asn Pro Leu Phe His 690 695 700 Gln Ala Ala Ser Arg Val Pro Ala Lys Gly Gly Ala Pro Ala Pro Ser 705 710 715 720 Arg Gly Pro Gln Glu Leu Val Pro Thr Thr His Pro Gly Gln Pro Ala 725 730 735 Arg His Pro Ala Ser Ser Val Ala Leu Lys Arg Pro Pro Pro Ala Pro 740 745 750 Pro Val Thr Val Ser Ser Pro Pro Phe Pro Val Pro Val Tyr Thr Arg 755 760 765 Gln Ala Pro Lys Gln Val Ile Lys Pro Thr Phe Ala Pro Pro Val Pro 770 775 780 Pro Val Lys Pro Gly Ala Gly Ala Ala Asn Pro Gly Pro Ala Glu Gly 785 790 795 800 Ala Val Gly Pro Lys Val Ala Leu Lys Pro Pro Ile Gln Arg Lys Gln 805 810 815 Gly Ala Gly Ala Pro Thr Ala Pro 820 <210> 2 <211> 702 <212> PRT <213> Homo sapiens <400> 2 Met Glu Ser Pro Ser Ala Pro Pro His Arg Trp Cys Ile Pro Trp Gln 1 5 10 15 Arg Leu Leu Leu Thr Ala Ser Leu Leu Thr Phe Trp Asn Pro Pro Thr 20 25 30 Thr Ala Lys Leu Thr Ile Glu Ser Thr Pro Phe Asn Val Ala Glu Gly 35 40 45 Lys Glu Val Leu Leu Leu Val His Asn Leu Pro Gln His Leu Phe Gly 50 55 60 Tyr Ser Trp Tyr Lys Gly Glu Arg Val Asp Gly Asn Arg Gln Ile Ile 65 70 75 80 Gly Tyr Val Ile Gly Thr Gln Gln Ala Thr Pro Gly Pro Ala Tyr Ser 85 90 95 Gly Arg Glu Ile Ile Tyr Pro Asn Ala Ser Leu Leu Ile Gln Asn Ile 100 105 110 Ile Gln Asn Asp Thr Gly Phe Tyr Thr Leu His Val Ile Lys Ser Asp 115 120 125 Leu Val Asn Glu Glu Ala Thr Gly Gin Phe Arg Val Tyr Pro Glu Leu 130 135 140 Pro Lys Pro Ser Ile Ser Ser Asn Asn Ser Lys Pro Val Glu Asp Lys 145 150 155 160 Asp Ala Val Ala Phe Thr Cys Glu Pro Glu Thr Gln Asp Ala Thr Tyr 165 170 175 Leu Trp Trp Val Asn Asn Gln Ser Leu Pro Val Ser Pro Arg Leu Gln 180 185 190 Leu Ser Asn Gly Asn Arg Thr Leu Thr Leu Phe Asn Val Thr Arg Asn 195 200 205 Asp Thr Ala Ser Tyr Lys Cys Glu Thr Gln Asn Pro Val Ser Ala Arg 210 215 220 Arg Ser Asp Ser Val Ile Leu Asn Val Leu Tyr Gly Pro Asp Ala Pro 225 230 235 240 Thr Ile Ser Pro Leu Asn Thr Ser Tyr Arg Ser Gly Glu Asn Leu Asn 245 250 255 Leu Ser Cys His Ala Ala Ser Asn Pro Ala Gln Tyr Ser Trp Phe 260 265 270 Val Asn Gly Thr Phe Gln Gln Ser Thr Gln Glu Leu Phe Ile Pro Asn 275 280 285 Ile Thr Val Asn Asn Ser Gly Ser Tyr Thr Cys Gln Ala His Asn Ser 290 295 300 Asp Thr Gly Leu Asn Arg Thr Thr Val Thr Thr Ile Thr Val Tyr Ala 305 310 315 320 Glu Pro Pro Lys Pro Phe Ile Thr Ser Asn Asn Ser Asn Pro Val Glu 325 330 335 Asp Glu Asp Ala Val Ala Leu Thr Cys Glu Pro Glu Ile Gln Asn Thr 340 345 350 Thr Tyr Leu Trp Trp Val Asn Asn Gln Ser Leu Pro Val Ser Pro Arg 355 360 365 Leu Gln Leu Ser Asn Asp Asn Arg Thr Leu Thr Leu Leu Ser Val Thr 370 375 380 Arg Asn Asp Val Gly Pro Tyr Glu Cys Gly Ile Gln Asn Glu Leu Ser 385 390 395 400 Val Asp His Ser Asp Pro Val Ile Leu Asn Val Leu Tyr Gly Pro Asp 405 410 415 Asp Pro Thr Ile Ser Pro Ser Tyr Thr Tyr Tyr Arg Pro Gly Val Asn 420 425 430 Leu Ser Leu Ser Cys His Ala Ala Ser Asn Pro Pro Ala Gln Tyr Ser 435 440 445 Trp Leu Ile Asp Gly Asn Ile Gln Gln His Thr Gln Glu Leu Phe Ile 450 455 460 Ser Asn Ile Thr Glu Lys Asn Ser Gly Leu Tyr Thr Cys Gln Ala Asn 465 470 475 480 Asn Ser Ala Ser Gly His Ser Arg Thr Thr Val Lys Thr Ile Thr Val 485 490 495 Ser Ala Glu Leu Pro Lys Pro Ser Ile Ser Ser Asn Asn Ser Lys Pro 500 505 510 Val Glu Asp Lys Asp Ala Val Ala Phe Thr Cys Glu Pro Glu Ala Gln 515 520 525 Asn Thr Thr Tyr Leu Trp Trp Val Asn Gly Gln Ser Leu Pro Val Ser 530 535 540 Pro Arg Leu Gln Leu Ser Asn Gly Asn Arg Thr Leu Thr Leu Phe Asn 545 550 555 560 Val Thr Arg Asn Asp Ala Arg Ala Tyr Val Cys Gly Ile Gln Asn Ser 565 570 575 Val Ser Ala Asn Arg Ser Asp Pro Val Thr Leu Asp Val Leu Tyr Gly 580 585 590 Pro Asp Thr Pro Ile Ile Ser Pro Asp Ser Ser Tyr Leu Ser Gly 595 600 605 Ala Asn Leu Asn Leu Ser Cys His Ser Ala Ser Asn Pro Ser Pro Gln 610 615 620 Tyr Ser Trp Arg Ile Asn Gly Ile Pro Gln Gln His Thr Gln Val Leu 625 630 635 640 Phe Ile Ala Lys Ile Thr Pro Asn Asn Asn Gly Thr Tyr Ala Cys Phe 645 650 655 Val Ser Asn Leu Ala Thr Gly Arg Asn Asn Ser Ile Val Lys Ser Ile 660 665 670 Thr Val Ser Ala Ser Gly Thr Ser Pro Gly Leu Ser Ala Gly Ala Thr 675 680 685 Val Gly Ile Met Ile Gly Val Leu Val Gly Val Ala Leu Ile 690 695 700

Claims (23)

agents that measure the expression level of ADAM8 (A Disintegrin and metalloproteinase domain-containing protein 8) or a gene encoding it; and
A composition for diagnosis of gastric cancer, comprising an agent for measuring the expression level of cancer embryonic antigen (CEA) or a gene encoding it,
When the expression level of ADAM8 measured in the biological sample of the subject of interest is 20 ng/ml or more, and the expression level of the cancer embryonic antigen is 3 ng/ml or more, it is predicted that the possibility of the gastric cancer is high, for diagnosis of gastric cancer composition. According to claim 1,
The biological sample is a liquid biopsy, a diagnostic composition for gastric cancer. 3. The method of claim 2,
The liquid biopsy is blood, serum or plasma, a composition for diagnosis of gastric cancer. delete According to claim 1,
The agent for measuring the expression level of the ADAM8 or the cancer embryonic antigen is an antibody, oligopeptide, ligand, PNA (peptide nucleic acid) and an aptamer that specifically binds to the ADAM8 or the cancer embryonic antigen. A composition for diagnosis of gastric cancer, comprising at least one selected from According to claim 1,
The agent for measuring the expression level of the ADAM8 or the gene encoding the cancer embryonic antigen is at least one selected from the group consisting of a primer, a probe and an antisense nucleotide that specifically binds to the ADAM8 or the gene encoding the cancer embryonic antigen. A composition for diagnosis of gastric cancer, comprising a. According to claim 1,
The composition is for diagnosing whether or not the onset of early stage gastric cancer or the possibility of onset, the composition for diagnosis of gastric cancer. According to claim 1,
The composition is for diagnosing whether or not the onset of N-stage gastric cancer or the possibility of the onset, the composition for diagnosis of gastric cancer. delete A kit for diagnosing gastric cancer comprising the composition for diagnosing gastric cancer according to any one of claims 1 to 3 and 5 to 8,
When the expression level of ADAM8 measured in the biological sample of the subject of interest is 20 ng/ml or higher and the expression level of the cancer embryonic antigen is 3 ng/ml or higher, it is predicted that the possibility of the gastric cancer is high, a kit for diagnosing gastric cancer . measuring the expression level of ADAM8 or a gene encoding the same in a biological sample isolated from a subject of interest; and
Measuring the expression level of a cancer embryonic antigen (CEA) or a gene encoding it in a biological sample isolated from the subject of interest
When the expression level of ADAM8 measured in the biological sample of the subject of interest is 20 ng/ml or more, and the expression level of the cancer embryonic antigen is 3 ng/ml or more, it is predicted that the likelihood of gastric cancer is high. A method of providing information regarding the diagnosis or staging of stomach cancer. 12. The method of claim 11,
The biological sample is a liquid biopsy, a method for diagnosing gastric cancer or providing information on the stage of gastric cancer. 13. The method of claim 12,
Wherein the liquid biopsy is blood, serum or plasma, a method for diagnosing gastric cancer or providing information on the stage of gastric cancer. delete 12. The method of claim 11,
The gastric cancer is an early stage gastric cancer, a method for diagnosing gastric cancer or providing information on the stage of gastric cancer. 12. The method of claim 11,
The stage of gastric cancer is an N-stage (N-stage) regarding whether lymph node metastasis or not, a method for diagnosing gastric cancer or providing information on the stage of gastric cancer. delete delete delete delete delete delete delete

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