KR20200139002A - Composition for diagnosing cancer - Google Patents

Abstract

The present invention relates to: a composition which can predict cancer stages and predict the risk of cancer occurrence at an early stage by measuring the expression level of ADAM8 or a gene encoding the same using a liquid biopsy of a target individual; a kit; and a method of providing information for the prediction.

Description

Composition for diagnosing cancer {Composition for diagnosing cancer}

The present invention relates to a composition, a kit, and a method for providing information for predicting the possibility of cancer incidence can be predicted at an early stage from a liquid biopsy of an object of interest, and the stage of cancer can also be predicted.

Gastric cancer is a cancer that occurs worldwide in Korea, China, and Japan, and its incidence is low in Western countries such as the United States and Europe, but in Korea, gastric cancer is the number one cancer incidence rate, and the mortality rate is second only to lung cancer. When looking at the classification of gastric cancer, 95% of the total is adenocarcinoma arising from the gland cells of the mucous membrane of the stomach wall, and there are lymphomas occurring in the lymphatic system and gastrointestinal interstitial tumors occurring in the interstitial tissue.

Currently, the three main treatment methods for cancer are surgical treatment, drug therapy, and radiation therapy. Drug therapy has less pain associated with treatment, and cancer recurrence is more likely to occur after treatment compared to surgical treatment or radiation therapy. As it is relatively small, expectations are raised about this. Therefore, numerous anticancer drugs have been developed and used to meet this, and most of these anticancer agents show anticancer effects by selectively killing actively dividing cells with the abnormal growth of cancer in mind. These anticancer drugs generally have a serious side effect of killing normal cells such as immune cells and hair follicle cells, which are cells actively dividing in the human body, so they have a problem that it cannot be used for a long time. In addition, methods for gastric cancer treatment include lymph node dissection, endoscopic mucosal resection, and laparoscopic gastrectomy. In endoscopic mucosal resection, physiological saline solution is applied around the gastric mucosa with cancer lesions for early gastric cancer in the mucous membrane. This is a method of convexly inflating the lesion area by injection and then resecting the mucous membrane with the lesion.It has the advantage of avoiding the pain of gastrectomy with a simple endoscopic procedure, but only in cases where the possibility of lymph node metastasis is low among early gastric cancers limited to the mucous membrane. There is a limit to use.

Recently, research is being conducted to discover therapeutic targets and use them for diagnosis and development of therapeutic agents through research on the function of genes related to cancer generation and treatment worldwide. Along with the activation of genomic research, human gene DNA chip or proteome analysis research has been actively conducted, and as a large number of cancer-related genes have been discovered, a list of many genes and related databases have been established. The biological function and cancer relevance are not yet studied or uncertain, and thus there is considerable difficulty in discovering a gene that can effectively treat cancer, along with actual cancer relevance or diagnosis and utilization as a target gene. Therefore, it is necessary to discover new genes in addition to cancer-related genes that have been discovered so far.

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

Another object of the present invention is to provide a method of providing information in order to diagnose the possibility of onset of cancer early 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 problem to be achieved by the present invention is not limited to the problems mentioned above, and other problems that are 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 diagnosis of cancer, comprising an agent measuring the expression level of ADAM8 (A Disintegrin and metalloproteinase domain-containing protein 8) or a gene encoding the same.

In the present invention, the "ADAM8" is an enzyme encoded by the ADAM8 gene, belongs to a membrane-anchored protein structurally related to snake venom disintegrins, and between cells-cells and cell-matrix It is involved in a number of biological processes. In addition, ADAM8 has been known to be involved in cell adhesion during neurodegeneration. In the present invention, 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 ADAM8 or a gene encoding it may be measured from a liquid biopsy of a target individual. 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 diagnosis of cancer, a part of a tissue in which cancer has occurred has been isolated and the expression level of the biomarker in the cancer tissue has been measured. Therefore, in the early stages of cancer, since it is difficult to take tissue due to difficulty in self-awareness, there is a disadvantage in that initial diagnosis is almost impossible, as well as the hassle of separating cells from the tissue.

However, in the present invention, not only can early diagnosis of cancer but also the stage of cancer can be predicted by measuring the expression level of ADAM8 or the gene encoding it in a liquid biopsy of blood, serum or plasma isolated from the target individual. have.

The composition of the present invention may further include an agent for measuring the expression level of a carcinoembryonic 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 gastrointestinal tissues during fetal development, and its production stops before birth. In the present invention, the cancer embryonic 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 ADAM8 or cancer embryonic antigen is not particularly limited, but preferably an antibody, oligopeptide, ligand, PNA (peptide nucleic acid) that specifically binds to the ADAM8 or cancer embryonic antigen protein. acid) and at least one selected from the group consisting of aptamer.

In the present invention, the "measurement of protein expression level" refers to a process of confirming the presence and expression of a protein of ADAM8 or a cancer embryo antigen, which is a diagnostic marker for cancer, in a biological sample in order to diagnose cancer. The ADAM8 or cancer embryonic antigen expression level measurement or comparison analysis methods include protein chip analysis, immunoassay, ligand binding assay, Matrix Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry (MALDI-TOF) analysis, and SELDI-TOF ( Sulface Enhanced Laser Desorption/Ionization Time of Flight Mass Spectrometry) analysis, radioimmunoassay, radioactive immunity diffusion method, octeroni immune diffusion method, rocket immunoelectrophoresis, tissue immunostaining, complement fixation analysis, 2D electrophoresis analysis, liquid chromatography Liquid chromatography-Mass Spectrometry (LC-MS), LC-MS/MS (liquid chromatography-Mass Spectrometry/ Mass Spectrometry), Western blotting and ELISA (enzyme linked immunosorbentassay), 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 cancer embryonic antigen. The antibodies of the present invention include polyclonal antibodies, monoclonal antibodies and recombinant antibodies. The antibody can be easily prepared using techniques well known in the art. For example, the polyclonal antibody can be produced by a method well known in the art, including the process of injecting the ADAM8 or the antigen of the cancer embryonic antigen into an animal and collecting blood from the animal to obtain serum containing the antibody. . Such polyclonal antibodies can be prepared from any animal such as goat, rabbit, sheep, monkey, horse, pig, cow, dog. In addition, the monoclonal antibody is a hybridoma method well known in the art (hybridoma method; see Kohler and Milstein (1976) European Journal of Immunology 6:511-519), or 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 a method such as gel electrophoresis, dialysis, salt precipitation, ion exchange chromatography, and affinity chromatography. In addition, the antibody of the present invention includes a complete form having two full-length light chains and two full-length heavy chains, as well as functional fragments of antibody molecules. The functional fragment of an antibody molecule means a fragment that has at least an antigen-binding function, and includes Fab, F(ab'), F(ab')2, and Fv.

In the present invention, the "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 Copenhagen University in Denmark in 1991. While DNA has a phosphate-ribose sugar backbone, PNA has a repeated N-(2-aminoethyl)-glycine backbone linked by a peptide bond, which greatly increases the binding power and stability to DNA or RNA, resulting in molecular biology. , Diagnostic analysis 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 oligonucleotide or a peptide molecule, and general information 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 embryo antigen is composed of a primer, a probe and an antisense nucleotide that specifically binds to the gene encoding the ADAM8 or 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 on this information.

In the present invention, the "measurement of the expression level of a gene" means measuring the amount of a gene by confirming the presence and expression level of the gene encoding ADAM8 or cancer embryonic antigen in the liquid biopsy to diagnose cancer. . Analysis 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), and RNase protection assay (RPA; RNase protection). assay), Northern blotting, DNA chip, etc., but are 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, preferably, a pair of primers that provides an analysis result having specificity and sensitivity. When the nucleic acid sequence of the primer is a sequence that is inconsistent with the non-target sequence present in the sample, a primer that amplifies only the target gene sequence containing the complementary primer binding site and does not induce non-specific amplification can give high specificity. .

In the present invention, the "probe" means 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 may be a peptide nucleic acid (PNA), a locked nucleic acid (LNA), a peptide, a polypeptide, a protein, RNA, or DNA, and most preferably For example, it may be PNA. More specifically, the probe is a biomaterial that includes an organism-derived or similar thing or a thing produced 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, oligonucleotide, RNA includes genomic RNA, mRNA, oligonucleotide, and examples of proteins include antibodies, antigens, enzymes, peptides, and the like.

In the present invention, the term "LNA (Locked nucleic acids)" refers to a nucleic acid analog including 2'-O, 4'-C methylene bridges [J Weiler, J Hunziker and J Hall Gene Therapy (2006) 13, 496.502 ]. LNA nucleosides contain common nucleic acid bases of DNA and RNA, and can form base pairs according to the Watson-Crick base pairing rules. However, due to the'locking' of the molecule due to the methylene bridge, the LNA cannot form an ideal shape in the Watson-Crick bond. When LNA is included in a DNA or RNA oligonucleotide, the LNA can more quickly pair with a complementary nucleotide chain to increase the stability of the double helix. In the present invention, the "antisense" refers to a sequence of nucleotide bases in which the antisense oligomer is hybridized with the target sequence in RNA by Watson-Crick base pairing, and typically allows the formation of mRNA and RNA: oligomer heterodimer within the target sequence. And an oligomer having a backbone between subunits. Oligomers may have exact sequence complementarity or approximate complementarity to the target sequence.

In the present invention, the "diagnosis" refers to determining the susceptibility of a subject to a specific disease or disease, determining whether the subject currently has a specific disease or disease, or having a specific disease or disease. Determining the subject's prognosis (e.g., identification of a pre-metastatic or metastatic cancer state, determining the stage of the cancer or determining the responsiveness of the cancer to treatment), or therametrics (e.g., for treatment efficacy Monitoring the state of an object to provide information). For the purposes of the present invention, the diagnosis is to determine whether or not the above-described disease is onset or the likelihood (risk), the stage of the cancer, or the survival rate or treatment responsiveness of the cancer patient.

In the present invention, the "stage" refers to the extent to which cancer cells have spread and the stage of progression of cancer, and the international classification according to the progression 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 metastasis to other organs. Corresponds to. The detailed classification in T, N, and M is shown in Table 1 below.

TNM weapon Justice Size of the primary tumor
(T weapon)
Size of the primary tumor(T stage) T0 Tumor cells that show the appearance of malignant tumors, but are limited to the mucous membrane or epithelium, and have not yet infiltrated the basement membrane. T1 Limited lesions in the primary organ, tumors are movable, and there is no invasion of adjacent and surrounding tissues. T2 The size of the tumor is about 2~5cm T3 The size of the tumor is larger than T2, but localized within the organ T4a Adhesion and infiltration with serosa tissue T4b Adhesion and infiltration with surrounding tissues Whether lymph node metastases
(N weapon)
Lymph node status(N stage) N0 No evidence of lymph node lesions N1 Invasion of a single, palpable, mobile, limited lymph node (1-2 cm or more, usually up to 3 cm in size) in the first position N2 Palpated, partially mobile or hard or hard lymph nodes with evidence of microscopic involvement, clinically entangled, appearing on opposite or bilateral sides (3-5 cm) N3 Complete?? It is fixed and completely fixed to bones, large blood vessels, skin, nerves, etc. through the film, and the size of more than 6cm Remote metastasis
(M weapon)
Distant metastasis
(M stage) M0 No distant metastasis M1 There is distant metastasis

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

In the present invention, ADAM8 from a liquid biopsy of a subject of interest, and optionally a cancer embryonic antigen; Alternatively, by measuring the expression level of a gene encoding them, it is possible to diagnose whether or not the onset of cancer in an early stage or the likelihood of developing it.

In addition, in the present invention, ADAM8, and optionally, cancer embryonic antigens 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 onset of N-stage cancer or the likelihood of developing cancer, or which stage the progression of cancer belongs to in the N-stage.

As a disease to be diagnosed in the present invention, the "cancer" represents or refers to a physiological condition characterized by uncontrolled cell growth typically in mammals. Cancers subject to diagnosis in the present invention are stomach cancer, ovarian cancer, colon 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 lymphoma , Hodgkin's lymphoma, hematologic cancer, bladder cancer, kidney cancer, melanoma, colon cancer, bone cancer, skin cancer, head cancer, uterine cancer, rectal cancer, brain tumor, cancer of the anus, fallopian tube carcinoma, endometrial carcinoma, vaginal cancer, vulvar carcinoma, esophageal cancer, small intestine Cancer, endocrine cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, ureteral cancer, renal cell carcinoma, renal pelvic carcinoma, CNS central nervous system tumor, primary CNS lymphoma, spinal cord tumor, brain stem glioma or pituitary gland It may be 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 using the diagnostic kit to diagnose the onset or possibility of developing cancer in an early stage, early diagnosis of cancer is possible, and the stage of cancer, preferably N-stage, can be predicted. .

In the present invention, the disease targeted for the diagnosis is gastric cancer, ovarian cancer, colon 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, hematologic cancer, bladder cancer, kidney cancer, melanoma, colon cancer, bone cancer, skin cancer, head cancer, uterine cancer, rectal cancer, brain tumor, near-anal 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, ureteral cancer, renal cell carcinoma, renal pelvic carcinoma, CNS central nervous system tumor, primary CNS lymphoma, spinal cord tumor, brainstem nerve It may include gliomas 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 kit for diagnosing cancer of the present invention may further include one kind or more other component compositions, solutions, or devices suitable for an analysis method.

For example, in the present invention, the kit for diagnosing cancer may further include essential elements necessary to perform a reverse transcription polymerase reaction. The reverse transcription polymerase reaction kit contains a pair of primers specific for the gene encoding the 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. In addition, a primer specific to the nucleic acid sequence of the control gene may be included. Other reverse transcription polymerase reaction kits include test tubes or other suitable containers, reaction buffers (various pH and magnesium concentrations), deoxynucleotides (dNTPs), enzymes such as Taq-polymerase and reverse transcriptase, DNase, RNase inhibitor DEPC. -May include DEPC-water, sterilized water, etc.

In addition, the diagnostic kit of the present invention may include essential elements necessary to perform 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. In addition, the substrate may include a cDNA or oligonucleotide corresponding to a control gene or a fragment thereof.

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

According to another embodiment of the present invention, regarding 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 target individual. will be.

In the present invention, the "object of interest" refers to an individual whose onset of the cancer is uncertain and has a high probability of developing the cancer.

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

In the present invention, it may include the step of measuring the expression level of ADAM8 or a gene encoding it in the biological sample isolated as described above.

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

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

In the present invention, as a method for measuring or comparing the expression level of the ADAM8 or cancer embryonic antigen, protein chip analysis, immunoassay, ligand binding assay, Matrix Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry (MALDI-TOF) analysis, SELDI-TOF (Sulface Enhanced Laser Desorption/Ionization Time of Flight Mass Spectrometry) analysis, radioimmunoassay, radioactive immunity diffusion method, octeroni immunity diffusion method, rocket immunoelectrophoresis, tissue immunostaining, complement fixation analysis method, 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 immunosorbentassay), 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 gene encoding the ADAM8 or cancer embryo antigen. It may include more than one.

In the present invention, as an analysis method for measuring the amount of a gene by confirming the presence and expression of the gene encoding the ADAM8 or cancer embryonic antigen, reverse transcription polymerase reaction (RT-PCR), 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 an individual of interest in the present invention; Alternatively, it may include the step of predicting the likelihood of developing cancer or the stage of the cancer by comparing the expression level of the gene encoding them with the level measured in the normal control group.

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

In addition, in the present invention, the cancer stage may be an N-stage regarding whether or not lymph node metastasis is present.

ADAM8, and optionally cancer embryonic antigen, measured against a biological sample of an individual 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 possibility 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 object 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, It can be predicted that the likelihood of developing it is high.

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

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

In the present invention, the cancer is gastric cancer, ovarian cancer, colon 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 lymphoma, Hodgkin lymphoma , Blood cancer, bladder cancer, kidney cancer, melanoma, colon cancer, bone cancer, skin cancer, head cancer, uterine cancer, rectal cancer, brain tumor, anal muscle 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, ureteral cancer, renal cell carcinoma, renal pelvic carcinoma, central nervous system tumor, primary CNS lymphoma, spinal cord tumor, brainstem glioma or pituitary adenoma. , Preferably it may be stomach cancer.

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

Contacting the biological sample with a test substance; And

It relates to a method for screening a drug for treating cancer comprising the step of measuring the expression level of the ADAM8 or the gene in the biological sample after contact with 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 for 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 decreased compared to prior to contact with the test substance, the step of determining the test substance as a therapeutic agent for cancer may be further included.

Further, 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 decreased compared to prior to contact with the test substance, the step of determining the test substance as an inhibitor of lymph node metastasis of cancer may be further included.

In the present invention, the object, biological sample and the definition of cancer, ADAM8 or cancer embryonic antigen; Alternatively, the method of measuring the expression level of the gene encoding them is duplicated as described in the method for providing information for diagnosis of cancer of the present invention, and thus detailed description thereof will be omitted in order to avoid excessive complexity of the specification.

According to another embodiment of the present invention, the step of treating a candidate substance predicted to induce cancer in the isolated biological sample; And

It relates to a method for screening a drug that induces cancer, comprising measuring the expression level of ADAM8 or a gene encoding it 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 for the biological sample isolated as described above, the accuracy in screening for the drug inducing the cancer is improved. You can increase it.

In the present invention, the candidate substances include 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 substance, the step of determining the candidate substance as a cancer inducing agent may be further included.

In addition, in the present invention, the ADAM8, and optionally, a cancer embryonic antigen (CEA) in the biological sample after treatment of 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 step of determining the candidate substance as a lymph node metastasis inducing agent of cancer may be further included.

In the present invention, the object, biological sample and the definition of cancer, ADAM8 or cancer embryonic antigen; Alternatively, the method of measuring the expression level of the gene encoding them is duplicated as described in the method for providing information for diagnosis of cancer of the present invention, and thus detailed description thereof will be omitted in order to avoid excessive complexity of the specification.

In the present invention, cancer can be diagnosed early by measuring the expression level of ADAM8 or a gene encoding the same for a liquid biopsy of a target individual, and the N-stage of cancer can be predicted. Compared to the method of measuring the expression level of a biomarker, it has the advantage of being non-invasive, simple, fast, but very accurate.

1 shows AUC and ROC curves of ADAM8 and CEA in blood for predicting gastric cancer 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 contents of the present invention are not limited by the following examples.

Example

Subject

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

A total of 80 individuals were selected as the initial training dataset, of which 20 were in the normal control group, 10 in the dysplastic group, 20 in the EGC group, 20 in the AGC group without metastasis, and the AGC group with metastasis. 10 were registered.

A total of 241 individuals were selected as the independent validation dataset, of which 70 were in the normal control group, 24 in the dysplastic group, 70 in the EGC group, 50 in the AGC group without metastasis, and the AGC group with metastasis. 27 people were registered.

Gastrointestinal endoscopy for all patients who visited Yonsei University Health System for gastric symptoms or routine examination in order to identify new markers for diagnosis of gastric cancer and stage diagnosis of gastric cancer that can improve diagnosis accuracy. endoscopy) (Types XQ-260, Olympus, Tokyo, Japan) was performed, and blood samples were obtained before further treatment. All gastric cancer patient groups were subjected to chest X-ray, abdominal-pelvic spiral CT scan, and whole body positron emission tomography (PET) for TNM staging. The stage of gastric 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 experiment. Patients with other gastric tumors such as other carcinomas or gastrointestinal stromal tumors, mucosa-associated lymphoid tissue lymphoma, and neuroendocrine tumors were also excluded from this study. In addition, patients who had previously received 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); Measurement of the expression level 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 of 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 then reacted at 37° C. for 1 hour. After washing each well three 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 to 20 minutes. Thereafter, the expression level of ADAM8 in the blood was measured at 450 nm using a VERSA max microplate reader (Molecular Devices Co., Sunnyvale, Calif., USA).

The expression level of CEA in blood was measured using Beckman Access CEA assay (Beckman Coulter Inc. Chaska, USA). Blood levels of IL-23, SDF-1 alpha, IL-8 and sCD40L were determined using a commercially available MILLIPLEX MAP Human Cytokine/Chemokine kit (Millipore, Billerica, MA, USA) using chemiluminescent immunoassay. It was measured using. Specifically, a filter plate was pre-immersed in 200 μl of an assay buffer at room temperature for 10 minutes, and then the assay buffer was vacuum removed. 25 μl of the standard or control solution was added to the wells and 25 μl of Assay Buffer was added to the sample wells and 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 sample was added. After mixing, 25 μl of beads were added, and the plate was shaken and incubated overnight at 4°C. After incubation, the fluid was removed and the plate was washed twice. After the detection antibody (25 μl) was added, the plate was incubated at room temperature for 1 hour. Streptavidin-phycoerythrin (25 μl) was added to each well containing 25 μl of a detection antibody, and then incubated for 1 hour at room temperature by shaking. After removing the fluid, the plate was washed, and 150 μl of sheath fluid was added. Luminex 100 after re-suspending for 5 minutes?? After reading the intermediate fluorescence intensity using IS, analysis was performed using a logistic curve-fitting method to determine the chemokine concentration.

Statistical analysis

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

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

Information of a total of 80 individuals was registered for the initial training data. The expression levels of ADAM8 and CEA in blood during gastric cancer carcinogenesis were measured, and the expression levels were compared in a total of five groups: the normal control group, the dysplastic group, the EGC group, the AGC group without metastasis, and the AGC group with metastasis. 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 different (ANOVA, p <0.001), and the expression level increased as the cancerous process progressed: normal/gastritis In the group, the expression level of ADAM8 in blood was 1.7±2.6 ng/mL, in the dysplastic group 15.3±13.3 ng/mL, in the EGC group 53.9±36.9 ng/mL, in the AGC group without metastasis, 82.8±52.6 ng/mL, primary AGC group with metastasis measured 53.6±26.3 ng/mL. It was confirmed that the level of ADAM8 expression in blood was remarkably increased in the early stages of gastric cancer. As shown in Table 3, the expression level of ADAM8 in blood in the early gastric cancer patients and EGC group was in the dysplasia group (high-risk, post-hoc Bonferroni, p <0.001) or the normal control group (control, post-hoc Bonferroni, p It was confirmed that it was significantly higher than <0.001). 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 group of advanced gastric cancer patients with primary metastasis, but the expression level was rather decreased. In addition, as shown in Table 4, as a result of comparing the average value of ADAM8 expression levels in blood between the cancer patient group and the non-cancer patient group, the cancer patient group was 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 blood was not significantly different between each group except for the AGC group with metastasis, and was significantly increased only in the AGC group with metastasis (ANOVA, p = 0.013). . As a result of comparing the mean expression level of CEA in blood 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 group
(30) Cancer patients
(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

Independent verification data were used to confirm the reproducibility of the results of the earlier training data. In the verification data, information on a total of 241 individuals was registered. Similar to the training data, the mean value of ADAM8 expression levels in blood increased with progression of cancer. According to Table 5, the expression level of ADAM8 in blood in the normal/gastritis group was 6.4±8.7 ng/mL, 23.7±16.0 ng/mL in the dysplastic group, 54.5±42.7 ng/mL in the EGC group, and the AGC group without metastasis. 67.9±55.1 ng/mL, AGC group with primary metastasis was measured to be 55.0±29.7 ng/mL (ANOVA, p <0.001). In the independent verification data, the expression level of ADAM8 in the blood significantly increased in the early gastric cancer stage, and the expression level of ADAM8 in the blood slightly increased in the advanced gastric cancer stage without metastasis compared to the early gastric cancer stage, but there was a significant difference between cancer patients. 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) was significantly higher than that of the blood ADAM8 expression level (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 significantly increased only in the metastatic AGC group, 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 group
(94) Cancer patients
(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 verification data

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

Clinical pathological characteristics Serum ADAM8 g _s ( p- value) Age (years) -0.064 (0.319) Gender Male Female) -0.095 (0.143) Tissue (well differentiated: medium 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-weapon (T1a:T1b:T2:T3:T4) 0.197 (0.085) N-Army (N0:N1:N2:N3) 0.320 (0.011) Primary metastasis (M0:M1) 0.140 (0.193)

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

As a result of confirming the ROC curves and measured AUC values of ADAM8 and CEA in blood for prediction of gastric cancer in the verification data, as shown in FIG. 1, the AUC of blood ADAM8 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), and it was found that ADAM8 in blood predicted the presence of gastric cancer, which was more accurate than blood CEA. 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 increased pattern of ADAM8 and CEA in the blood was remarkably different in cancer progression, it was predicted that there would be a synergistic effect when using both markers 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 these 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 verification data

In order to confirm the role of ADAM8 in blood in the process of gastric cancer, correlation between the expression level of ADAM8 in blood and pre-inflammatory or pro-angiogenic cytokines related to gastric cancer in verification 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). ), whereas there was no significant correlation between blood IL-8 (γp = 0.113, p = 0.313) and blood sCD40L (γp = 0.043, p = 0.702), an angiogenic cytokine. Could From this, it was found that ADAM8 in the blood was associated with cancer-related inflammation or local invasion 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 diagnosing 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 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 Gln 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 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 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)

ADAM8 (A Disintegrin and metalloproteinase domain-containing protein 8) or a composition for diagnosis of cancer comprising an agent for measuring the expression level of a gene encoding the same. The method of claim 1,
The expression level of the ADAM8 or the gene encoding it is measured from a liquid biopsy of a target individual, a composition for diagnosis of cancer. The method of claim 2,
The liquid biopsy is blood, serum or plasma, a composition for diagnosis of cancer. The method of claim 1,
The composition further comprises an agent for measuring the expression level of a cancer embryonic antigen (CEA) or a gene encoding the same. The method of claim 1,
The agent for measuring the expression level of ADAM8 comprises at least one selected from the group consisting of antibodies, oligopeptides, ligands, peptide nucleic acids (PNA) and aptamers that specifically bind to ADAM8. A composition for diagnosis of. The method of claim 1,
The agent for measuring the expression level of the gene encoding ADAM8 comprises at least one selected from the group consisting of primers, probes, and antisense nucleotides specifically binding to the gene encoding ADAM8. The method of claim 1,
The composition is for diagnosing the onset or possibility of the onset of cancer in the early stage (early stage), a composition for diagnosis of cancer. The method of claim 1,
The composition is for diagnosing the onset or possibility of the onset of cancer of the N-stage (N-stage), a composition for diagnosis of cancer. The method of claim 1,
The cancer is gastric cancer, ovarian cancer, colon 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 lymphoma, Hodgkin lymphoma, blood cancer. , Bladder cancer, kidney cancer, melanoma, colon cancer, bone cancer, skin cancer, head cancer, uterine cancer, rectal cancer, brain tumor, anal muscle 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, ureteral cancer, renal cell carcinoma, renal pelvic carcinoma, central nervous system tumor, primary CNS lymphoma, spinal cord tumor, brain stem glioma or pituitary adenoma, composition for diagnosis of cancer . A kit for diagnosis of cancer comprising the composition for diagnosis of any one of claims 1 to 9. A method for diagnosing cancer or providing information on the stage of cancer, comprising the step of measuring the expression level of ADAM8 or a gene encoding the same in a biological sample isolated from a target individual. The method of claim 11,
The biological sample is a liquid biopsy, a method for providing information on the diagnosis or stage of cancer. The method of claim 12,
The liquid biopsy is blood, serum, or plasma, a method of providing information on the diagnosis or stage of cancer. The method of claim 11,
Diagnosis of cancer, further comprising the step of predicting the likelihood of onset or stage of cancer by comparing the expression level of ADAM8 or the gene encoding the same with the level measured in the normal control group, measured for the biological sample of the object of interest Or a method of providing information about the stage of cancer. The method of claim 14,
The cancer is an early stage cancer, a method of providing information on the diagnosis or stage of cancer. The method of claim 14,
The cancer stage is an N-stage with respect to lymph node metastasis, a method for diagnosing cancer or providing information on the stage of cancer. The method of claim 14,
When the expression level of ADAM8 or the gene encoding the same measured for the biological sample of the object of interest is increased compared to the normal control, the diagnosis of cancer or the diagnosis of cancer comprising the step of predicting that the onset of the cancer is high How to provide information about the weapon. The method of claim 14,
When the expression level of ADAM8 measured in the biological sample of the object of interest is 20 ng/ml or more, it is predicted that the possibility of developing the cancer is high. The method of claim 11,
The information providing method further comprises the step of measuring the expression level of a cancer embryonic antigen (CEA) or a gene encoding the same with respect to the separated biological sample, diagnosis of cancer or providing information on the stage of cancer. The method of claim 19,
When the expression level of ADAM8 measured in the biological sample of the object of interest is 20 ng/ml or more and the expression level of the cancer embryonic antigen is 3 ng/ml or more, the cancer is predicted to have a high probability of developing the cancer. How to provide information about the diagnosis or stage of cancer. The method of claim 11,
The cancer is gastric cancer, ovarian cancer, colon 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 lymphoma, Hodgkin lymphoma, blood cancer. , Bladder cancer, kidney cancer, melanoma, colon cancer, bone cancer, skin cancer, head cancer, uterine cancer, rectal cancer, brain tumor, anal muscle 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, ureteral cancer, renal cell carcinoma, renal pelvic carcinoma, central nervous system tumor, primary CNS lymphoma, spinal cord tumor, brain stem glioma or pituitary adenoma, diagnosis of cancer or How to provide information about the stage of cancer. Measuring the expression level of ADAM8 or a gene encoding it in the isolated biological sample;
Contacting the biological sample with a test substance; And
A method for screening a drug for treating cancer comprising the step of measuring the expression level of the protein or the gene in the biological sample after contacting the test substance. Treating the isolated biological sample with a candidate substance predicted to induce cancer; And
A method for screening a drug that induces cancer, comprising measuring the expression level of ADAM8 or a gene encoding it in a biological sample treated with the candidate substance.

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