JP2008014937A - Tumor marker and method for determination of occurrence of cancerous disease - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a tumor marker for colorectal cancer; and a method for the determination of the occurrence of colorectal cancer. <P>SOLUTION: The tumor marker comprising a protein identified in a colorectal cancer tissue is provided. The method for the determination of the occurrence of colorectal cancer using the tumor marker is provided. In the method, the level of the protein in a sample derived from an individual who is a target of the determination of the occurrence of colorectal cancer is measured, and the measured level is compared with the normal level for the protein. The increase or decrease in the measured level relative to the normal level is employed as an indicator of the possibility that the individual is suffering from colorectal cancer. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to techniques for clinical diagnosis / examination / follow-up observation and classification, and more specifically to a tumor marker and a method for identifying the onset of a cancer disease. In particular, the present invention relates to a tumor marker for colorectal cancer and a method for identifying morbidity of colorectal cancer.

  Blood tests are performed as one of the methods for diagnosis, screening, and follow-up of colorectal cancer. In blood tests, the presence of advanced cancer can be estimated by measuring the concentration of a certain protein (tumor marker) in the blood of a patient. The tumor marker for colorectal cancer is described in, for example, Anticancer Research, 2004, 24 (4), 2519-2530 (Non-patent Document 1). Currently, typical tumor markers include carcinoembryonic antigen (CEA) and CA19-9. When there is metastasis to the liver or the like, the concentration of these tumor markers is highly elevated. In addition, US2006 / 0019256 (COMPOSITIONS AND METHODS FOR TREATING AND DIAGNOSING CANCER) (Patent Document 1) includes HLA-C, MBC2, RAB22A, MGC15429, MHC class I as analysis results of cultured cells derived from lung cancer (xenograft tumors). Antigen is described as a cancer-related protein. Furthermore, US2003 / 0211498 (TUMOR MARKERS IN OVARIAN CANCER) (Patent Document 2) describes Tumor rejection antigen-1 as a uterine cancer marker gene.

“Anticancer Research”, 2004, Vol. 24, No. 4, p. 2519-2530 US Patent Application Publication No. 2006/0019256 US Patent Application Publication No. 2003/0211498

  As described above, tumor markers such as carcinoembryonic antigen (CEA) and CA19-9 are used in blood tests for diagnosis, screening, and follow-up of colorectal cancer. However, the concentration of these tumor markers is often at a normal level in the early cancer stage, and is often in the normal level range even in the advanced cancer stage. From such a point of view, there is a demand for the discovery of a novel tumor marker that has high specificity for the affected site and is suitable for early diagnosis.

  Note that Patent Document 1 does not describe that the disclosed cancer-related protein is related to colorectal cancer. Further, Patent Document 2 does not describe that the disclosed uterine cancer marker gene is related to colorectal cancer.

  An object of the present invention is to provide a tumor marker with higher specificity for colorectal cancer. Another object of the present invention is to provide a method that can identify the morbidity of colorectal cancer.

  The present inventors used pathological tissue and normal tissue in the colonic mucosa epithelium collected from patients with colorectal cancer as samples, and used the NBS method as a proteome analysis method, and a difference of a certain amount or more is recognized between both tissues. Protein groups were identified. The NBS method is an excellent technique for relative quantification of proteins and peptides using 2-nitrobenzenesulfenyl chloride. The proteome analysis method using the NBS method is fundamentally different from the most general-purpose proteome analysis method using the two-dimensional electrophoresis method in the separation mode and the detection principle. Therefore, according to the proteome analysis method using the NBS method, it is considered that a marker candidate protein different from the protein group reported as a renal cancer marker candidate can be found so far.

The present invention includes the following inventions.
The following (1) and (2) relate to tumor markers for colorectal cancer.

The following (1) relates to a tumor marker for colorectal cancer containing a protein whose expression level is higher than the normal level in cancer patients.
(1)
6-phosphogluconolactonase, Alpha1 acid glyco protein, Alpha-actinin 1, Apurinic endonuclease, Calumenin, Chaperonin, Clathrin heavy chain 1, Clathrin light polypeptide A, c-myc binding protein, Complement factor H, Cysteine rich intestinal protein 1, F-box protein 40, Fibrinogen gamma, Fk506 Binding Protein Fkbp Mutant R42kH87V COMPLEX WITH Immunosuppressant Fk506, Guanine nucleotide binding protein (G protein), beta polypeptide 2-like 1, Heat shock 70kD protein 9B, Heparan sulfate proteoglycan 2, HLA-C, hypothetical protein FLJ38663, L-plastin polypeptide, MBC2, Migration inhibitory factor-related protein 14 variant E, Mitogen inducible gene, Proteasome subunit p58, RAB18, member RAS oncogene family, RAB22A, Radixin, RAN, member RAS oncogene family, Rhodanese; thiosulfate sulfurtransferase, Ribosomal protein L13, Ribosomal protein L27a, Ribosomal protein L4, Ribosomal protein S18, Ribosomal protein S29, Ribosome binding protein 1, Sadenosylhomocystei ne hydrolase, Solute carrier family 25 (mitochondrial carrier; adenine nucleotide translocator), member 5, Solute carrier family 3 (activator of dibasic and neutral amino acid transport), member 2, Splicing factor 3B, subunit 3, Splicing factor, arginine / serine -rich 3 (SRp20), U5 snRNP-specific protein, 116 kD, Ubiquitin isopeptidase T, Vitronectin, XTP3 transactivatied protein A, Galectin 1, Reticulocalbin 1, Vimentin, ESP-2 (zyxin); Protein tyrosine phosphatase receptor type C, Protein Large intestine containing at least one protein selected from the group consisting of tyrosine phosphatase, receptor type, alpha, orosomucoid 2, Tumor rejection antigen 1, glycyl-tRNA synthetase, TLS protein, Ribonuclease RNase A family 3, and heterogeneous nuclear ribonucleoprotein H2. Tumor marker for cancer.

The following (2) relates to a tumor marker for colorectal cancer containing a protein whose expression level is small compared with the normal level in the colorectal cancer patient body.
(2)
ADP-ribosylation factor-like 10C, aldehyde dehydrogenase2, Alpha-actinin 4, Annexin A2 isoform 2, ATP synthase, H + transporting, mitochondrial F0 complex, subunit d isoform a, ATP-binding cassette transporter family A member 12, Calnexin, Carbonic Anhydrase Form B, Carbonyl reductase 1, Cathepsin S, cysteine rich protein 1, Dynein light chain 1, Endoplasmic-reticulum-lumenal protein 28, Enoyl coenzyme hydrase, short chain1, Eukaryotic translation elongation factor 2, Filamin B, gelsolin isoform a, Glucosamine- fructose-6-phosphate aminotransferase, GTP-binding protein Rab3B, Haptoglobin, Heterogeneous nuclear ribonucleoprotein A2, Hydroxymethylglutaryl-CoA synthase, mitochondrial, Isocitrate dehydrogenase 1, Lymphocyte cytosolic protein 1, Major vault protein, MGC15429 protein, MHCin I antigen heavy polypeptide 14, Myozenin 3, NADH Ubiquinone oxidoreductase subunit B13, Normal mucosa of esophagus specific 1, Olfactomedin 4, phosphoenolpyruvate calboxykinase 2, Phosphoglycerate mutase 1, Proline arginine-rich end leucine-rich repeat protein precursor, Protein kinase C and casein kinase substrate in neurons 2, Protein P97, Pyridoxine 5'-phosphate oxidase, Raf kinase inhibitor protein, Ras associated protein Rab5B, A tumor marker for colorectal cancer comprising a protein selected from the group consisting of Retinoblastoma binding protein 4, succinate dehydrogenase complex, subunit A, flavoprotein, Thioredoxin domain containing 5, TNRC15 protein; Collagen, type XIV, alpha 1, and Desmoglein 2.

  The following (3) to (6) relate to a method for identifying the incidence of colorectal cancer. In the present invention, morbidity widely refers to a pathological state, and identifying morbidity of colorectal cancer is used in the sense including detection, diagnosis, monitoring, staging, and prognosis determination of colorectal cancer.

  The following (3) and (4) relate to a method for identifying morbidity of colorectal cancer using the tumor marker for colorectal cancer described in (1) above.

(3)
6-phosphogluconolactonase, Alpha1 acid glyco protein, Alpha-actinin 1, Apurinic endonuclease, Calumenin, Chaperonin, Clathrin heavy chain 1, Clathrin light polypeptide A, c-myc binding protein, Complement factor H, Cysteine rich intestinal protein 1, F-box protein 40, Fibrinogen gamma, Fk506 Binding Protein Fkbp Mutant R42kH87V COMPLEX WITH Immunosuppressant Fk506, Guanine nucleotide binding protein (G protein), beta polypeptide 2-like 1, Heat shock 70kD protein 9B, Heparan sulfate proteoglycan 2, HLA-C, hypothetical protein FLJ38663, L-plastin polypeptide, MBC2, Migration inhibitory factor-related protein 14 variant E, Mitogen inducible gene, Proteasome subunit p58, RAB18, member RAS oncogene family, RAB22A, Radixin, RAN, member RAS oncogene family, Rhodanese; thiosulfate sulfurtransferase, Ribosomal protein L13, Ribosomal protein L27a, Ribosomal protein L4, Ribosomal protein S18, Ribosomal protein S29, Ribosome binding protein 1, Sadenosylhomocystei ne hydrolase, Solute carrier family 25 (mitochondrial carrier; adenine nucleotide translocator), member 5, Solute carrier family 3 (activator of dibasic and neutral amino acid transport), member 2, Splicing factor 3B, subunit 3, Splicing factor, arginine / serine -rich 3 (SRp20), U5 snRNP-specific protein, 116 kD, Ubiquitin isopeptidase T, Vitronectin, XTP3 transactivatied protein A, Galectin 1, Reticulocalbin 1, Vimentin, ESP-2 (zyxin); Protein tyrosine phosphatase receptor type C, Protein At least one protein selected from the group consisting of tyrosine phosphatase, receptor type, alpha, orosomucoid 2, Tumor rejection antigen 1, glycyl-tRNA synthetase, TLS protein, Ribonuclease RNase A family 3, and heterogeneous nuclear ribonucleoprotein H2 A method for identifying the incidence of colorectal cancer by using it as a tumor marker.

(4)
Measuring the level of the protein in a sample derived from an individual to be identified for colorectal cancer incidence;
Comparing the measured level obtained with the normal level of the protein,
The obtained measured value level is higher than the normal level, which is one of the indicators that the subject individual is likely to have colorectal cancer. To identify the incidence of colorectal cancer.

  The normal level of the protein may be the protein level in a non-cancerous sample serving as a control for a cancerous sample. For example, the level in a normal sample derived from a healthy individual, the level in a non-cancerous normal sample derived from a patient suffering from a cancer disease, and the like can be mentioned.

  The method according to (4), wherein the sample is serum or urine, and the level of the protein is measured by a test based on biospecific affinity.

  The following (5) and (6) relate to a method for identifying morbidity of colorectal cancer using the tumor marker for colorectal cancer described in (2) above.

(5)
ADP-ribosylation factor-like 10C, aldehyde dehydrogenase2, Alpha-actinin 4, Annexin A2 isoform 2, ATP synthase, H + transporting, mitochondrial F0 complex, subunit d isoform a, ATP-binding cassette transporter family A member 12, Calnexin, Carbonic Anhydrase Form B, Carbonyl reductase 1, Cathepsin S, cysteine rich protein 1, Dynein light chain 1, Endoplasmic-reticulum-lumenal protein 28, Enoyl coenzyme hydrase, short chain1, Eukaryotic translation elongation factor 2, Filamin B, gelsolin isoform a, Glucosamine- fructose-6-phosphate aminotransferase, GTP-binding protein Rab3B, Haptoglobin, Heterogeneous nuclear ribonucleoprotein A2, Hydroxymethylglutaryl-CoA synthase, mitochondrial, Isocitrate dehydrogenase 1, Lymphocyte cytosolic protein 1, Major vault protein, MGC15429 protein, MHCin I antigen heavy polypeptide 14, Myozenin 3, NADH Ubiquinone oxidoreductase subunit B13, Normal mucosa of esophagus specific 1, Olfactomedin 4, phosphoenolpyruvate calboxykinase 2, Phosphoglycerate mutase 1, Proline arginine-rich end leucine-rich repeat protein precursor, Protein kinase C and casein kinase substrate in neurons 2, Protein P97, Pyridoxine 5'-phosphate oxidase, Raf kinase inhibitor protein, Ras associated protein Rab5B, Retinoblastoma binding protein 4, succinate dehydrogenase complex, subunit A, flavoprotein, Thioredoxin domain containing 5, TNRC15 protein; a protein selected from the group consisting of Collagen, type XIV, alpha 1 and Desmoglein 2 is used as a tumor marker for colorectal cancer To identify the incidence of colorectal cancer.

(6)
Measuring the level of the protein in a sample derived from an individual to be identified for colorectal cancer incidence;
Comparing the measured level obtained with the normal level of the protein,
(6) The obtained measured value level is reduced from the normal level as one of indicators that the subject individual is likely to have colorectal cancer. To identify the incidence of colorectal cancer.

  The normal level of the protein may be the protein level in a non-cancerous sample serving as a control for a cancerous sample. For example, the level in a normal sample derived from a healthy individual, the level in a non-cancerous normal sample derived from a patient suffering from a cancer disease and the like can be mentioned.

  The method according to (6), wherein the sample is serum or urine, and the level of the protein is measured by a test based on biospecific affinity.

  The present invention is also directed to a pharmaceutical composition for the treatment of colorectal cancer described in (7) and (8) below. The treatment of colon cancer includes killing colon cancer cells and suppressing the growth of colon cancer cells.

(7)
A pharmaceutical composition for inducing a reaction that promotes death of colorectal cancer cells and / or suppression of colorectal cancer cell growth by supplying the colorectal cancer cells to the tumor marker according to (1) A pharmaceutical composition comprising at least one antibody that immunospecifically binds.

(8)
A pharmaceutical composition for promoting an immune response by supplying an immune stimulating amount to colon cancer cells, comprising the tumor marker according to (1).

  The pharmaceutical compositions of (7) and (8) above can be identified as potential therapeutic agents used in the treatment of colorectal cancer. Or the pharmaceutical composition of said (7) and (8) can be used as a therapeutic agent used for the treatment of colon cancer.

  According to the present invention, a tumor marker with higher specificity for colorectal cancer can be provided. Further, according to the present invention, it is possible to provide a method capable of identifying the morbidity of colorectal cancer.

<Tumor marker>
The present invention provides a tumor marker for colorectal cancer.
The protein provided as a tumor marker in the present invention is extracted from a cancerous part or a non-cancerous part of a colonic mucosa epithelial tissue collected from a colorectal cancer patient, and isotope labeled using 2-nitrobenzenesulfenyl chloride (NBSCl) ( (NBS method) and a proteome analysis technique based on a separation method using HPLC. In the NBS method, one of two kinds of protein samples is modified with a heavy reagent (2-nitro [ ¹³ C ₆ ] benzenesulfenyl chloride) and the other is modified with a light reagent (2-nitro [ ¹² C ₆ ] benzenesulfuric acid). The two NBS-modified protein samples were mixed with each other, subjected to appropriate treatment by those skilled in the art, such as trypsin digestion, and the difference in the amount of peptide was measured with a mass spectrometer. It is a method which can investigate the relative difference of the protein content in the protein sample. The NBS method is described in Rapid Commun. Mass Spectrom., 2003, 17, 1642-1650 and International Publication No. 2004/002950 pamphlet.

  Conventional two-dimensional electrophoresis methods have been difficult to separate and detect proteins having a basic isoelectric point and proteins having a large molecular weight of about 100 KDa. In the NBS method used in the present invention, peptide fragments obtained by trypsin digestion of proteins contained in tissues are directly analyzed, and HPLC is used for separation, so there are no restrictions on molecular weight and isoelectric point. . From this, there is a possibility of obtaining something different from the knowledge obtained conventionally.

  These proteins can be isolated and purified by any protein purification technique selected by one skilled in the art. For example, techniques including chromatography such as ion exchange, affinity, and size exclusion column chromatography, centrifugation, differential solubility, and electrophoresis can be used.

  Among the proteins provided by the present invention as a tumor marker, in the cancer patient body, as a protein whose expression level is higher than the normal level, 6-phosphogluconolactonase, Alpha1 acid glycoprotein, Alpha-actinin 1, Apurinic endonuclease, Calumenin, Chaperonin, Clathrin heavy chain 1, Clathrin light polypeptide A, c-myc binding protein, Complement factor H, Cysteine rich intestinal protein 1, F-box protein 40, Fibrinogen gamma, Fk506 Binding Protein Fkbp Mutant R42kH87V COMPLEX WITH Immunosuppressant Fk506, Guanine nucleotide binding protein (G protein), beta polypeptide 2-like 1, Heat shock 70kD protein 9B, Heparan sulfate proteoglycan 2, HLA-C, hypothetical protein FLJ38663, L-plastin polypeptide, MBC2, Migration inhibitory factor-related protein 14 variant E, Mitogen inducible gene, Proteasome subunit p58, RAB18, member RAS oncogene family, RAB22A, Radixin, RAN, member RAS oncogene family, Rhodanese; thiosulfate sulfurtransferase Ribosomal protein L13, Ribosomal protein L27a, Ribosomal protein L4, Ribosomal protein S18, Ribosomal protein S29, Ribosome binding protein 1, Sadenosylhomocysteine hydrolase, Solute carrier family 25 (mitochondrial carrier; adenine nucleotide translocator), member 5, Solute carrier family 3 ( activator of dibasic and neutral amino acid transport), member 2, Splicing factor 3B, subunit 3, Splicing factor, arginine / serine-rich 3 (SRp20), U5 snRNP-specific protein, 116 kD, Ubiquitin isopeptidase T, Vitronectin, XTP3 transactivatied protein A, Galectin 1, Reticulocalbin 1, Vimentin, ESP-2 (zyxin); Protein tyrosine phosphatase receptor type C, Protein tyrosine phosphatase, receptor type, alpha, orosomucoid 2, Tumor rejection antigen 1, glycyl-tRNA synthetase, TLS protein, Ribonuclease RNase A family 3 and heterogeneous nuclear ribonucleoprotein H2 are provided.

  For example, the protein has a tendency to show an expression level of about 50% or more of the number of colon cancer patients and about 50% or more, preferably 100% or more larger in the cancer part than in the non-cancer part.

  Furthermore, among the above proteins, Protein tyrosine phosphatase receptor type C, Protein tyrosine phosphatase, receptor type, alpha, orosomucoid 2, Tumor rejection antigen 1, glycyl-tRNA synthetase, TLS protein, Ribonuclease RNase A family 3, and heterogeneous nuclear ribonucleoprotein H2 Is specifically expressed in the cancerous part (or is below the detection limit in the non-cancerous part).

  Among the proteins provided by the present invention as a tumor marker, ADP-ribosylation factor-like 10C, aldehyde dehydrogenase2, Alpha-actinin 4, Annexin A2 isoform 2 , ATP synthase, H + transporting, mitochondrial F0 complex, subunit d isoform a, ATP-binding cassette transporter family A member 12, Calnexin, Carbonic Anhydrase Form B, Carbonyl reductase 1, Cathepsin S, cysteine rich protein 1, Dynein light chain 1, Endoplasmic-reticulum-lumenal protein 28, Enoyl coenzyme hydrase, short chain1, Eukaryotic translation elongation factor 2, Filamin B, gelsolin isoform a, Glucosamine-fructose-6-phosphate aminotransferase, GTP-binding protein Rab3B, Haptoglobin, Heterogeneous nuclear ribonucleoprotein A2, Hydroxymethylglutaryl-CoA synthase, mitochondrial, Isocitrate dehydrogenase 1, Lymphocyte cytosolic protein 1, Major vault protein, MGC15429 protein, MHC cla ss I antigen, Myosin, heavy polypeptide 14, Myozenin 3, NADH Ubiquinone oxidoreductase subunit B13, Normal mucosa of esophagus specific 1, Olfactomedin 4, phosphoenolpyruvate calboxykinase 2, Phosphoglycerate mutase 1, Proline arginine-rich end leucine-rich repeat protein precursor kinase C and casein kinase substrate in neurons 2, Protein P97, Pyridoxine 5'-phosphate oxidase, Raf kinase inhibitor protein, Ras associated protein Rab5B, Retinoblastoma binding protein 4, succinate dehydrogenase complex, subunit A ,, flavoprotein, Thioredoxin domain containing 5, Provide TNRC15 protein; Collagen, type XIV, alpha 1, and Desmoglein 2.

For example, the protein has a tendency to exhibit an expression level of about 50% or more of the number of colon cancer patients and higher in the non-cancerous part than in the cancerous part by about 50% or more, preferably 100% or more.
Furthermore, among the above proteins, Collagen, type XIV, alpha 1, and Desmoglein 2 are specifically expressed in the non-cancerous part (or are below the detection limit in the cancerous part).

<Method for identifying the incidence of colorectal cancer>
The present invention also provides a method for identifying the onset of colorectal cancer by using at least one protein selected from the above protein group as a tumor marker.

  In the method of the present invention, a sample derived from an individual who is to be identified as suffering from colorectal cancer is prepared, and the level of at least one protein selected from the protein group is measured in the sample. The obtained measurement level is compared with the normal level. The normal level is the protein level in a sample serving as a control for a cancerous sample of at least one protein selected from the above protein group. Here, the sample serving as a control for the cancerous sample is not particularly limited as long as it is a non-cancerous sample. For example, a sample derived from a healthy individual, a normal sample derived from a patient suffering from a cancer disease, and the like can be mentioned.

  In the present invention, a sample derived from an individual who is a subject to be identified as suffering from colorectal cancer is not particularly limited. Examples include cells, tissues, body fluids, and tissue extracts. The cells and tissues include tissue biopsy materials and autopsy materials, and also include tissue sections and tissue extracts thereof. In particular, examples of the sample include cells, tissues, and tissue extracts of large intestine-derived samples. Samples derived from the large intestine include mucosal epithelium, lamina propria, mucosal muscle plate, submucosa, lamina propria, serosa and the like. Body fluids include blood, urine, and body secretions. The blood includes whole blood, plasma, serum and the like. Tissue extract refers to tissue homogenated or solubilized by methods known to those skilled in the art. Among these exemplified samples, it is preferable to use serum and / or urine as a sample.

  Tumor marker measurements measured in a sample containing a subject's individual cells, tissues, body fluids, and tissue extracts, preferably non-cancerous samples containing the same type of cells, tissues, body fluids, and tissue extracts Compared to the level in

  In identifying the incidence of colorectal cancer, tumor markers are used in the following applications. Tumor markers in tissues are, for example, cancer diagnosis and prognosis using tissue sections (for example, performed by immunohistochemical staining, mass imaging, etc.), and PET diagnosis (for example, using radioactive probes for tumor markers) ) Or a drug target for treatment (see <Pharmaceutical Composition> described below). In addition, a tumor marker in blood is a target to be quantitatively measured in, for example, cancer diagnosis and prognosis.

  The above proteins are 6-phosphogluconolactonase, Alpha1 acid glycoprotein, Alpha-actinin 1, Apurinic endonuclease, Calumenin, Chaperonin, Clathrin heavy chain 1, Clathrin light polypeptide A, c-myc binding protein, Complement factor H, Cysteine rich intestinal protein 1, F-box protein 40, Fibrinogen gamma, Fk506 Binding Protein Fkbp Mutant R42kH87V COMPLEX WITH Immunosuppressant Fk506, Guanine nucleotide binding protein (G protein), beta polypeptide 2-like 1, Heat shock 70kD protein 9B, Heparan sulfate proteoglycan 2, HLA -C, hypothetical protein FLJ38663, L-plastin polypeptide, MBC2, Migration inhibitory factor-related protein 14 variant E, Mitogen inducible gene, Proteasome subunit p58, RAB18, member RAS oncogene family, RAB22A, Radixin, RAN, member RAS oncogene family, Rhodanese; thiosulfate sulfurtransferase, Ribosomal protein L13, Ribosomal protein L27a, Ribosomal protein L4, Ribosomal protein S18, Ribosomal protein S29, Ribosome binding pr otein 1, Sadenosylhomocysteine hydrolase, Solute carrier family 25 (mitochondrial carrier; adenine nucleotide translocator), member 5, Solute carrier family 3 (activator of dibasic and neutral amino acid transport), member 2, Splicing factor 3B, subunit 3, Splicing factor , arginine / serine-rich 3 (SRp20), U5 snRNP-specific protein, 116 kD, Ubiquitin isopeptidase T, Vitronectin, XTP3 transactivatied protein A, Galectin 1, Reticulocalbin 1, Vimentin, ESP-2 (zyxin); Protein tyrosine phosphatase receptor Measurements obtained in the group consisting of type C, protein tyrosine phosphatase, receptor type, alpha, orosomucoid 2, Tumor rejection antigen 1, glycyl-tRNA synthetase, TLS protein, Ribonuclease RNase A family 3, and heterogeneous nuclear ribonucleoprotein H2. The value level is higher than the normal level, which is an indicator that individuals who should be identified as having colorectal cancer are more likely to have colorectal cancer. It is possible to bracts. As a degree of increase in the measurement value level, it is good to use a measure that the measurement value level is 50% or more, preferably 100% or more of the normal level.

  Furthermore, among the above proteins, Protein tyrosine phosphatase receptor type C, Protein tyrosine phosphatase, receptor type, alpha, orosomucoid 2, Tumor rejection antigen 1, glycyl-tRNA synthetase, TLS protein, Ribonuclease RNase A family 3, and heterogeneous nuclear ribonucleoprotein H2 Is a specific protein in colorectal cancer tissue, and if its presence is confirmed, it is highly likely that the individual who should be identified as having colorectal cancer suffers from colorectal cancer. It can be one of the indicators.

  The above proteins are ADP-ribosylation factor-like 10C, aldehyde dehydrogenase2, Alpha-actinin 4, Annexin A2 isoform 2, ATP synthase, H + transporting, mitochondrial F0 complex, subunit d isoform a, ATP-binding cassette transporter family A member 12 , Calnexin, Carbonic Anhydrase Form B, Carbonyl reductase 1, Cathepsin S, cysteine rich protein 1, Dynein light chain 1, Endoplasmic-reticulum-lumenal protein 28, Enoyl coenzyme hydrase, short chain1, Eukaryotic translation elongation factor 2, Filamin B, gelsolin isoform a, Glucosamine-fructose-6-phosphate aminotransferase, GTP-binding protein Rab3B, Haptoglobin, Heterogeneous nuclear ribonucleoprotein A2, Hydroxymethylglutaryl-CoA synthase, mitochondrial, Isocitrate dehydrogenase 1, Lymphocyte cytosolic protein 1, Major vault protein, MGC15429 I antigen, Myosin, heavy polypeptide 14, Myozenin 3, NADH Ubiquinone oxidoreductase subunit B13, Normal mucosa of esophagus specific 1, Olfact omedin 4, phosphoenolpyruvate calboxykinase 2, Phosphoglycerate mutase 1, Proline arginine-rich end leucine-rich repeat protein precursor, Protein kinase C and casein kinase substrate in neurons 2, Protein P97, Pyridoxine 5'-phosphate oxidase, Raf kinase inhibitor protein, Ras Associated protein Rab5B, Retinoblastoma binding protein 4, succinate dehydrogenase complex, subunit A,, flavoprotein, Thioredoxin domain containing 5, TNRC15 protein; Collagen, type XIV, alpha 1 and Desmoglein 2 If the level is lower than the normal level, it can be one of the indicators that an individual who is to be identified as suffering from colorectal cancer is likely to have colorectal cancer. As a degree of decrease in the measurement value level, it is preferable that the normal level is 50% or more, preferably 100% or more of the measurement value level.

  Furthermore, among the above proteins, Collagen, type XIV, alpha 1 and Desmoglein 2 are specific proteins in normal tissues (non-cancerous tissues), so if their absence is confirmed, colon cancer is affected. It can be one of the indicators that the individual to be identified is likely to have colorectal cancer.

  The protein level is preferably measured by a test based on biospecific affinity. The test based on biospecific affinity is a method well known to those skilled in the art, and is not particularly limited, but an immunoassay is preferable. Specifically, Western blot, radioimmunoassay, ELISA, sandwich immunoassay, immunoprecipitation method, precipitation reaction, in-gel diffusion sedimentation reaction, immunodiffusion method, aggregation measurement, complement binding assay, immunoradiometric assay, fluorescent immunoassay, Immunoassays are included, including competitive and non-competitive assay systems, such as protein A immunoassays. In an immunoassay, the presence of an antibody that binds to a tumor marker in a sample of an individual is detected. Specifically, it is carried out by contacting a sample with the antibody in an assay medium under conditions capable of forming an immune complex consisting of a tumor marker protein to be measured and an antibody of the protein. More specific immunoassay protocols can be easily selected by those skilled in the art.

  The protein level is preferably measured by a test based on biospecific affinity as described above, but can also be measured by other protein quantification methods. For example, the NBS method already described is an excellent method for quantitativeness. In this case, using a sample prepared with a known level of the above protein or an appropriate sample such as a normal sample as a control sample, the measurement is performed by examining the difference in the abundance of the protein from the sample of the subject individual. It can be carried out.

  In the method of the present invention, the tumor marker of the present invention may be measured alone or in combination with any other tumor marker. Accordingly, the methods of the present invention may include measuring the levels of other tumor markers as well as the levels of the tumor markers of the present invention.

  The tumor marker of the present invention is preferably used for the purpose of detection, diagnosis, monitoring, staging and prognosis determination of colorectal cancer. Preferably, it may be used for grasping the dynamics of the tumor. For example, when chemotherapy or radiation therapy is performed on a tumor, it may be used to determine how effective the treatment is. Moreover, when excision surgery is performed on a tumor having a high tumor marker value, it may be used for post-operative follow-up.

<Pharmaceutical composition>
The present invention is also directed to a pharmaceutical composition for treating colorectal cancer by using a tumor marker comprising a protein whose expression level is higher than a normal level in a colon cancer patient, among the tumor markers of the present invention. .

  One form of the pharmaceutical composition is a pharmaceutical composition for inducing a response that promotes death of cancer cells and / or suppression of growth of cancer cells by supplying the cancer composition to the cancer cells. A pharmaceutical composition comprising at least one antibody that immunospecifically binds to a marker. Antibodies include polyclonal antibodies, monoclonal antibodies, and antibodies prepared by molecular biological techniques. Here, the antibody may be any substance that binds widely immunospecifically, and an antibody fragment or an antibody fusion protein may also be used. In either case, antibody preparation is carried out by methods well known to those skilled in the art.

  Another form of the pharmaceutical composition is a pharmaceutical composition for promoting an immune response by supplying an immune stimulating amount to a cancer cell, the pharmaceutical composition comprising the tumor marker of the present invention. It is. Here, the immunostimulatory amount refers to the amount of an antigen capable of eliciting a desired immune response for the treatment of cancer, and is determined by a method well known by those skilled in the art. According to this form, the cancer is treated using a method well known to those skilled in the art, known as so-called cancer vaccine therapy.

  The pharmaceutical composition contains the antibody or tumor marker as an active ingredient, and may further contain a pharmaceutically acceptable diluent, carrier, excipient, and the like. The pharmaceutical composition can be identified as a potential therapeutic agent used in the treatment of colorectal cancer. Alternatively, the pharmaceutical composition can be applied as a therapeutic agent used for the treatment of colorectal cancer.

  EXAMPLES Hereinafter, the present invention will be specifically described with reference to examples. However, the present invention is not limited to the following examples.

[Example 1: Protein expression analysis by NBS method]
Normal tissues (ie, non-cancerous part of the colonic mucosal epithelial tissue) and cancerous tissues (ie, cancerous part of the colonic mucosal epithelial tissue) were collected from patients with colorectal cancer. Each tissue was homogenized in solubilization buffer A (50 mM Tris-HCl (pH 8.0), 100 mM NaCl, 10 mM EDTA, protease inhibitor (aprotinin, PMSF, leupepsin) aqueous solution), and 100000 × G (4 ° C , 1 hour), and the supernatant was obtained as a soluble fraction. Next, the precipitate obtained by centrifugation was homogenized again in solubilization buffer B (9 M urea, 2 w / v% CHAPS, 10 mM EDTA, protease inhibitor (aprotinin, PMSF, leupepsin aqueous solution)), and 100000 × G ( After ultracentrifugation at 4 ° C. for 1 hour, the supernatant was obtained as an insoluble fraction.

The extracted proteins of these soluble and insoluble fractions were each treated with NBS reagent, and protein expression analysis was performed. Regarding NBS reagent treatment, stable isotope labeling was performed according to the recommended protocol of ¹³ C NBS Stable Isotope Labeling Kit-N (Shimadzu Corporation). Specifically, proteins extracted from normal tissues were labeled with ¹² CNBS (Light NBS), and proteins extracted from cancer tissues were labeled with ¹³ CNBS (Heavy NBS). Each of the obtained labeled proteins was mixed, and desalted, reduced, alkylated, and trypsin digested according to the recommended protocol of the kit. Next, the labeled peptide was concentrated according to the protocol of the kit. Concentrated labeled peptides were subsequently separated by μHPLC using a C18 column and applied to MS plates. The sample thus prepared was measured using a mass spectrometer Axima-CFR plus (manufactured by Shimadzu Corporation), and a relative quantitative analysis of each protein was performed.

  The peptide fragment showing a pair peak was subjected to MS / MS analysis using a tandem mass spectrometer Axima-QIT (manufactured by Shimadzu Corporation) to identify the protein.

  By using such a method, the same analysis was finally performed on 24 clinical specimens. As a result of analysis, in 50% or more of the total number of samples, a protein in which the expression level of the protein in the cancer tissue was increased or decreased was identified. The degree of increase or decrease was based on the following amount. That is, in a cancer part, a peptide having an increased abundance in a non-cancer part has an abundance in the cancer part of 50% or more than that in the non-cancer part (that is, the abundance in the cancer part is non- Those with 150% or more of the existing amount in the cancerous part) were selected. In addition, in the cancerous part, the peptide whose abundance in the non-cancerous part is reduced is more than 50% greater than the abundance in the non-cancerous part (the abundance in the cancerous part is non-cancerous). Less than 66% of the abundance in the part).

  As a result, the proteins shown in Tables 1 to 8 below were identified. In the table, “Mean”, “SEM”, and “SD” are relative ratios (%) measured in each clinical specimen (that is, cancer when the protein expression level in the non-cancerous part is 100 (%)). The average value, standard error, and standard deviation value of the expression level (ratio)) are shown.

  Tables 1 to 3 are a list of proteins (Up-regulated proteins) that increase the expression level of the protein in the cancerous part than in the non-cancerous part. For example, proteins such as 6-phosphogluconolactonase (average expression level: about 2.16 times that of non-cancerous part) whose expression level increased in 16 out of 24 samples were identified. Table 4 is a list of proteins that were specifically detected in the cancer site (All or None). “150% ≦” in Tables 1 to 4 indicates the number of specimens whose abundance in the cancerous part was 150% or more of the abundance in the non-cancerous part.

  Tables 5 to 7 are lists of proteins (Down-regulated proteins) in which the expression level of the protein in the cancerous part is reduced rather than in the non-cancerous part. For example, proteins such as Carbon Anhydrase Form B (average expression level: 0.11 times the non-cancerous part) whose expression level decreases in 15 samples out of 24 samples were identified. Table 8 is a list of proteins specifically detected in the non-cancerous part (All or None). “<66%” in Tables 5 to 8 indicates the number of specimens in which the abundance in the cancerous part was less than 66% of the abundance in the non-cancerous part.

  In Examples 2 and 3 below, among the proteins that showed an increase in the cancerous part than in the non-cancerous part by the NBS method, ZYX (Zyxin), RAN (RAN, member RAS oncogene family), RCN1 (Reticulocalbin), Six types of proteins, AHCY (S-adenosylhomocysteine hydrolase), SGALS1 (Galectin1), and VIM (Vimentin), were subjected to Western blotting analysis of biological tissue and validation analysis by immunohistochemical staining of biological tissue.

[Example 2: Western blotting analysis]
Normal tissues (that is, non-cancerous portions of the colonic mucosa epithelial tissue) and cancer tissues (that is, cancerous portions of the colonic mucosal epithelial tissue) collected from colon cancer patients (5 cases) were prepared. Each tissue was homogenized in solubilization buffer A (50 mM Tris-HCl (pH 8.0), 100 mM NaCl, 10 mM EDTA, protease inhibitor (aprotinin, PMSF, leupepsin) aqueous solution), and 100000 × G (4 ° C , 1 hour), and the supernatant was obtained as a soluble fraction. Next, the precipitate obtained by centrifugation was homogenized again in solubilization buffer B (9M urea, 2% CHAPS, 10 mM EDTA, protease inhibitor (aprotinin, PMSF, leupepsin) aqueous solution), and 100000 × G (4 ° C , 1 hour), and the supernatant was obtained as an insoluble fraction.

  Among these protein extraction solutions, soluble fractions are numbered with patients 1-5, and each is used for analysis of ZYX, RAN, RCN1, and AHCY, insoluble fractions are numbered with patients 6-10, Used for analysis of SGALS1 and VIM, respectively.

  Each extract was separated by 12.5% acrylamide (only 15% acrylamide sample used for analysis of SGALS1) by SDS-PAGE gel, and then transferred to a nitrocellulose membrane. The transferred sample was loaded with a primary antibody (described later) for each protein and incubated at room temperature for 2 hours. After the reaction, the nitrocellulose membrane was washed with a PBS (Phosphate Buffered Saline) buffer, a secondary antibody (described later) solution was added, and the mixture was reacted for 1 hour. After reacting with the antibody, each band was detected by adding an ECL coloring solution.

The antibodies used in this example are as follows.
Primary antibody:
Mouse anti-human ZYX polyclonal antibody (antibody dilution ratio 1/500)
Mouse anti-human RAN monoclonal antibody (antibody dilution ratio 1/2000)
Mouse anti-human AHCY polyclonal antibody (antibody dilution ratio 1/1000)
Rabbit anti-human RCN1 monoclonal antibody (antibody dilution ratio 1/1000)
Rabbit anti-human SGAL1 polyclonal antibody (antibody dilution ratio 1/1000)
Rabbit anti-human VIM polyclonal antibody (antibody dilution ratio 1/1000)

Secondary antibody:
Horseradish peroxidase conjugate-sheep anti-mouse IgG antibody (antibody dilution ratio 1/400)
Horseradish peroxidase conjugate-donkey anti-rabbit IgG antibody (antibody dilution ratio 1/400)

FIG. 1 shows an electrophoretogram obtained by the above Western blotting. In FIG. 1, N indicates a non-cancerous part-derived sample, and T indicates a cancerous part-derived sample.
As shown in FIG. 1, it was confirmed that the expression of all six types of proteins was enhanced in all five cases of cancer. That is, in the quantification result by the NBS method shown in Example 1 above, the protein that showed increased expression in the cancer part than in the non-cancer part actually showed such increased expression in the quantification result by Western blotting. . Thus, the quantitative result by the NBS method and the quantitative result by Western blotting show a good correlation. Therefore, it was confirmed that these molecules are proteins related to colorectal cancer.

  From this, the protein that showed increased expression in the non-cancerous part from the cancer part in the quantification result by the NBS method, that is, the protein that showed the expression suppression in the cancerous part from the non-cancerous part, It is easily estimated that such an expression tendency is shown.

[Example 3: Analysis by immunohistochemical staining method]
Paraffin-embedded sections (4 μm) of normal tissue (ie, non-cancerous part of colonic mucosal epithelial tissue) and cancerous tissue (ie, cancerous part of colonic mucosal epithelial tissue) collected from colon cancer patients (10 cases) were prepared. . As the antibody, the same antibody as that used in the Western blotting analysis was used.

  The paraffin section and the primary antibody were reacted for 1 hour at room temperature, washed with PBS buffer, and further reacted with streptavidin-biotin peroxidase complex. Finally, tissue staining was performed by reacting the tissue section with a color-developing solution (3,3'-diaminobenzydynetetrahydrochloride, 0.01% peroxidase, 0.05 M Tris aqueous solution (pH 7.6)) for 3 minutes. It was. The result is shown in FIG.

In FIG. 2, (A) shows ZYX, (B) shows RAN, (C) shows RCN1, (D) shows AHCY, (E) shows SGALS1, and (F) shows immunohistochemical staining results for VIM. , T indicates the staining result in the cancerous part, and N indicates the staining result in the non-cancerous part.
As shown in FIG. 2, in all cases, it was confirmed that the expression of the protein in all the six types of proteins was enhanced in cancer cells (or stromal cells surrounding the cancer cells) in the cancerous part. That is, in the results of quantification by the NBS method shown in Example 1 above, proteins that showed increased expression in the cancerous part than in the non-cancerous part actually showed such enhanced expression in the analysis result by the immunohistochemical staining method. Was showing. Thus, the quantitative result by the NBS method and the analysis result by the immunohistochemical staining method show a good correlation. Therefore, it was confirmed that these molecules are proteins related to colorectal cancer.

  From this, the analysis results by immunohistochemical staining of the proteins that showed increased expression in the non-cancerous part from the cancer part in the quantification result by the NBS method, that is, the proteins that showed the expression suppression in the cancerous part from the non-cancerous part In this case, it is easily estimated that such an expression tendency is shown.

  From the above, the protein of claim 1 is expressed more in colorectal cancer tissue than in non-cancerous tissue, and the protein of claim 3 is more expressed than in non-cancerous tissue. Less expressed in colorectal cancer tissues. It is clear that these proteins can be used as tumor markers for colorectal cancer. When these proteins are used as tumor markers for colorectal cancer and the colorectal cancer disease is identified for a sample whose colorectal cancer disease is unknown, the methods described in Examples 1 to 3 can be used.

  In the above-described embodiments, specific forms within the scope of the present invention have been shown, but the present invention is not limited to these and can be carried out in various other forms. For this reason, the said Example is only a mere illustration in all points, and must not be interpreted limitedly. Further, all modifications belonging to the equivalent scope of the claims are within the scope of the present invention.

The protein (6 types) whose expression is enhanced in the colorectal cancer tissue in the present invention shows the results of Western blotting analysis in clinical samples (cancerous and non-cancerous parts of colonic mucosal epithelial tissue of colorectal cancer patients). FIG. 4 shows the results of validation analysis by immunohistochemical staining of clinical samples (cancerous and non-cancerous parts of colorectal mucosal epithelial tissue of colorectal cancer patients) for proteins (six types) whose expression is enhanced in the colorectal cancer tissues in the present invention.

Claims (6)

  6-phosphogluconolactonase, Alpha1 acid glyco protein, Alpha-actinin 1, Apurinic endonuclease, Calumenin, Chaperonin, Clathrin heavy chain 1, Clathrin light polypeptide A, c-myc binding protein, Complement factor H, Cysteine rich intestinal protein 1, F-box protein 40, Fibrinogen gamma, Fk506 Binding Protein Fkbp Mutant R42kH87V COMPLEX WITH Immunosuppressant Fk506, Guanine nucleotide binding protein (G protein), beta polypeptide 2-like 1, Heat shock 70kD protein 9B, Heparan sulfate proteoglycan 2, HLA-C, hypothetical protein FLJ38663, L-plastin polypeptide, MBC2, Migration inhibitory factor-related protein 14 variant E, Mitogen inducible gene, Proteasome subunit p58, RAB18, member RAS oncogene family, RAB22A, Radixin, RAN, member RAS oncogene family, Rhodanese; thiosulfate sulfurtransferase, Ribosomal protein L13, Ribosomal protein L27a, Ribosomal protein L4, Ribosomal protein S18, Ribosomal protein S29, Ribosome binding protein 1, Sadenosylhomocystei ne hydrolase, Solute carrier family 25 (mitochondrial carrier; adenine nucleotide translocator), member 5, Solute carrier family 3 (activator of dibasic and neutral amino acid transport), member 2, Splicing factor 3B, subunit 3, Splicing factor, arginine / serine -rich 3 (SRp20), U5 snRNP-specific protein, 116 kD, Ubiquitin isopeptidase T, Vitronectin, XTP3 transactivatied protein A, Galectin 1, Reticulocalbin 1, Vimentin, ESP-2 (zyxin); Protein tyrosine phosphatase receptor type C, Protein Large intestine containing at least one protein selected from the group consisting of tyrosine phosphatase, receptor type, alpha, orosomucoid 2, Tumor rejection antigen 1, glycyl-tRNA synthetase, TLS protein, Ribonuclease RNase A family 3, and heterogeneous nuclear ribonucleoprotein H2. Cancer marker.   ADP-ribosylation factor-like 10C, aldehyde dehydrogenase2, Alpha-actinin 4, Annexin A2 isoform 2, ATP synthase, H + transporting, mitochondrial F0 complex, subunit d isoform a, ATP-binding cassette transporter family A member 12, Calnexin, Carbonic Anhydrase Form B, Carbonyl reductase 1, Cathepsin S, cysteine rich protein 1, Dynein light chain 1, Endoplasmic-reticulum-lumenal protein 28, Enoyl coenzyme hydrase, short chain1, Eukaryotic translation elongation factor 2, Filamin B, gelsolin isoform a, Glucosamine- fructose-6-phosphate aminotransferase, GTP-binding protein Rab3B, Haptoglobin, Heterogeneous nuclear ribonucleoprotein A2, Hydroxymethylglutaryl-CoA synthase, mitochondrial, Isocitrate dehydrogenase 1, Lymphocyte cytosolic protein 1, Major vault protein, MGC15429 protein, MHCin I antigen heavy polypeptide 14, Myozenin 3, NADH Ubiquinone oxidoreductase subunit B13, Normal mucosa of esophagus specific 1, Olfactomedin 4, phosphoenolpyruvate calboxykinase 2, Phosphoglycerate mutase 1, Proline arginine-rich end leucine-rich repeat protein precursor, Protein kinase C and casein kinase substrate in neurons 2, Protein P97, Pyridoxine 5'-phosphate oxidase, Raf kinase inhibitor protein, Ras associated protein Rab5B, A colorectal cancer marker comprising a protein selected from the group consisting of Retinoblastoma binding protein 4, succinate dehydrogenase complex, subunit A, flavoprotein, Thioredoxin domain containing 5, TNRC15 protein; Collagen, type XIV, alpha 1, and Desmoglein 2.   6-phosphogluconolactonase, Alpha1 acid glyco protein, Alpha-actinin 1, Apurinic endonuclease, Calumenin, Chaperonin, Clathrin heavy chain 1, Clathrin light polypeptide A, c-myc binding protein, Complement factor H, Cysteine rich intestinal protein 1, F-box protein 40, Fibrinogen gamma, Fk506 Binding Protein Fkbp Mutant R42kH87V COMPLEX WITH Immunosuppressant Fk506, Guanine nucleotide binding protein (G protein), beta polypeptide 2-like 1, Heat shock 70kD protein 9B, Heparan sulfate proteoglycan 2, HLA-C, hypothetical protein FLJ38663, L-plastin polypeptide, MBC2, Migration inhibitory factor-related protein 14 variant E, Mitogen inducible gene, Proteasome subunit p58, RAB18, member RAS oncogene family, RAB22A, Radixin, RAN, member RAS oncogene family, Rhodanese; thiosulfate sulfurtransferase, Ribosomal protein L13, Ribosomal protein L27a, Ribosomal protein L4, Ribosomal protein S18, Ribosomal protein S29, Ribosome binding protein 1, Sadenosylhomocystei ne hydrolase, Solute carrier family 25 (mitochondrial carrier; adenine nucleotide translocator), member 5, Solute carrier family 3 (activator of dibasic and neutral amino acid transport), member 2, Splicing factor 3B, subunit 3, Splicing factor, arginine / serine -rich 3 (SRp20), U5 snRNP-specific protein, 116 kD, Ubiquitin isopeptidase T, Vitronectin, XTP3 transactivatied protein A, Galectin 1, Reticulocalbin 1, Vimentin, ESP-2 (zyxin); Protein tyrosine phosphatase receptor type C, Protein At least one protein selected from the group consisting of tyrosine phosphatase, receptor type, alpha, orosomucoid 2, Tumor rejection antigen 1, glycyl-tRNA synthetase, TLS protein, Ribonuclease RNase A family 3, and heterogeneous nuclear ribonucleoprotein H2 is a colorectal cancer marker. As a method of identifying the incidence of colorectal cancer by using as. Measuring the level of the protein in a sample derived from an individual to be identified for colorectal cancer incidence;
Comparing the measured level obtained with the normal level of the protein,
The obtained measured value level is higher than the normal level, which is one of the indicators that the subject individual is likely to have colorectal cancer. To identify the incidence of colorectal cancer.   ADP-ribosylation factor-like 10C, aldehyde dehydrogenase2, Alpha-actinin 4, Annexin A2 isoform 2, ATP synthase, H + transporting, mitochondrial F0 complex, subunit d isoform a, ATP-binding cassette transporter family A member 12, Calnexin, Carbonic Anhydrase Form B, Carbonyl reductase 1, Cathepsin S, cysteine rich protein 1, Dynein light chain 1, Endoplasmic-reticulum-lumenal protein 28, Enoyl coenzyme hydrase, short chain1, Eukaryotic translation elongation factor 2, Filamin B, gelsolin isoform a, Glucosamine- fructose-6-phosphate aminotransferase, GTP-binding protein Rab3B, Haptoglobin, Heterogeneous nuclear ribonucleoprotein A2, Hydroxymethylglutaryl-CoA synthase, mitochondrial, Isocitrate dehydrogenase 1, Lymphocyte cytosolic protein 1, Major vault protein, MGC15429 protein, MHCin I antigen heavy polypeptide 14, Myozenin 3, NADH Ubiquinone oxidoreductase subunit B13, Normal mucosa of esophagus specific 1, Olfactomedin 4, phosphoenolpyruvate calboxykinase 2, Phosphoglycerate mutase 1, Proline arginine-rich end leucine-rich repeat protein precursor, Protein kinase C and casein kinase substrate in neurons 2, Protein P97, Pyridoxine 5'-phosphate oxidase, Raf kinase inhibitor protein, Ras associated protein Rab5B, By using a protein selected from the group consisting of Retinoblastoma binding protein 4, succinate dehydrogenase complex, subunit A ,, flavoprotein, Thioredoxin domain containing 5, TNRC15 protein; Collagen, type XIV, alpha 1, and Desmoglein 2 as a colorectal cancer marker , A method to identify the incidence of colorectal cancer. Measuring the level of the protein in a sample derived from an individual to be identified for colorectal cancer incidence;
Comparing the measured level obtained with the normal level of the protein,
The obtained measured value level is reduced from the normal level as one of indicators that the subject individual is likely to have colorectal cancer. To identify the incidence of colorectal cancer.