WO2010100899A1 - Genetic testing method for cancer by analysis of expression of cancer-relating gene utilizing monocyte contained in blood sample - Google Patents

Abstract

Disclosed is a genetic testing method which can predict the risk of occurrence of a cancer cell mass or the efficacy of a therapy with high accuracy in an early stage or after prognosis. Specifically disclosed is a genetic testing method for cancer, which comprises: a step (10) of separating a buffy coat layer from a blood sample by centrifugation; a step (20) of separating a monocyte by density gradient centrifugation; a step (30) of extracting total RNA from the monocyte; a step (40) of synthesizing complementary DNA from the total RNA by reverse transcription reaction; a step (50) of carrying out a real-time-PCR (RT-PCR) analysis for a cancer-relating gene; and a step (60) of carrying out a DNA microarray analysis for a cancer-relating gene by employing a DNA microarray as a nucleic acid microarray.

Description

Methods for genetic testing of cancer by analyzing the expression of cancer-related genes using mononuclear cells contained in blood samples

The present invention relates to a cancer genetic testing method, and more particularly to a cancer genetic testing method based on an expression analysis of a cancer-related gene obtained from a mononuclear cell contained in a blood sample.

Cancer is one of the major lifestyle-related diseases and is the leading cause of death in Japan. There are cancers that develop and progress due to environmental factors and cancers that develop and develop due to genetic factors, and the onset factors and progression processes are diverse. In cancers that develop and progress due to genetic factors, the multistage carcinogenic mechanism of colorectal cancer is well known. Colorectal cancer progresses from a polyp (adenoma) state as a precancerous lesion to malignant colorectal cancer in multiple stages. This multi-stage cancer is first mutated in the RAS gene, which is one of the oncogenes, after the APC gene, a gene responsible for a dominant genetic disease called familial adenomatous adenomatosis, is inactivated and a polyp is generated. As a result, the cell proliferation action is activated, and then the p53 tumor suppressor gene is inactivated and progresses to malignant colon cancer (Non-patent Document 1). Such abnormal activation (overexpression) of oncogenes and inactivation (decrease in expression) of tumor suppressor genes that would normally suppress cell carcinogenesis are observed in the process of cell carcinogenesis. This is why cancer is said to be a genetic disease caused by accumulation of gene mutations, amplifications and deletions.

Currently, as diagnostic methods for cancer, for example, diagnostic imaging methods such as MRI (magnetic resonance imaging), PET (positron tomography), SPECT (single photon emission tomography), and X-ray CT (X-ray computed tomography) Is frequently used. These diagnostic imaging methods are basically methods for detecting cancer by finding the difference between normal tissue and diseased tissue, and can be said to be highly accurate cancer testing methods because the diseased tissue can be visually recognized. . In addition, a method for examining a cancer antigen (tumor marker) released in a blood sample using, for example, ELISA (enzyme immunoassay) is simple and widely used.

However, the detection level capable of image diagnosis in the above-described image diagnosis method is a cancer cell population that has developed to a size of 5 mm or more, and cancer cells that have reached this level are deteriorated levels associated with invasion and angiogenesis. . In addition, in the tumor marker test, the concentration of the target protein as a tumor marker in the blood sample needs to be high, and it is difficult to say that the test method is generally highly sensitive. Furthermore, in the tumor marker test, it is necessary to use an antibody in order to capture the target protein, and there is a possibility that a false positive occurs in terms of the specificity of the antibody.

Therefore, from the viewpoint of early detection and early treatment of cancer, an inspection method capable of detecting a cancer cell population at a detection level (5 mm or less) impossible with image diagnostic methods is desired. As described above, since cancer is accompanied by various gene abnormalities, it is considered that early detection and early treatment of cancer can be achieved by detecting these gene abnormalities.

The present invention has been made in view of the above circumstances, and an object of the present invention is to provide a genetic test method capable of predicting the existence risk of a cancer cell population and a therapeutic effect with high accuracy and early or prognosis. Is to provide.

As a result of diligent research, the inventors of the present invention have focused on mononuclear cells existing in human blood and have completed the present invention. That is, the cancer genetic testing method according to the present invention includes a step of separating mononuclear cells from a blood sample collected from a subject, a step of extracting total RNA from the separated mononuclear cells, and The method includes a step of synthesizing complementary DNA from total RNA and a step of performing expression analysis on a cancer-related gene in the synthesized complementary DNA.

Mononuclear cells existing in the blood are classified into migrating mononuclear leukocytes represented by lymphocytes and monocytes and mononuclear phagocytic cell groups represented by macrophages. Lymphocytes are mainly classified into B cells, T cells, and NK (natural killer) cells. B cells produce antibodies that bind to various antigens and attack pathogens that have entered from the outside. T cells are further classified into helper T cells that assist B cell antibody production, conversely suppressor T cells that suppress B cell antibody production, killer T cells that attach to pathogens and destroy them directly. NK cells are lymphocytes having a tumor lysis function. Monocytes do not contain special granules such as those found in granulocytes, but they have phagocytic ability and exhibit a variety of functions in biological defense reactions. Macrophages are thought to have been converted monocytes generated in the bone marrow and are also called phagocytic cells. Macrophages are strong in phagocytic digestion (phagocytosis) of foreign substances such as bacteria and cells that have undergone apoptosis, and are one of the main roles in immune function.

And these cells maintain their immune functions by emphasizing them while performing their functions. For example, macrophages fragment the foreign matter taken up by phagocytosis and present the antigen by binding this fragment to MHC-II on the cell surface. Then, antigen presentation by macrophages activates helper T cells and produces cytokines such as interleukins and lymphokines. Interleukins and cytokines produced by helper T cells activate macrophages and activate B cells that recognize the same antigen as the antigen presented. Activated B cells differentiate into antibody-producing cells and proliferate to produce antibodies against the antigen. The antibody produced by the antibody producing cell binds to the antigen to form an antibody-antigen complex. In this way, the foreign substance to which the antibody is bound is easily recognized by macrophages, and thus is efficiently phagocytosed.

As described above, for example, cancer cells that have induced apoptosis or necrosis are efficiently captured by the immune function maintained by the mononuclear cells. Therefore, in the present invention, since expression analysis of cancer-related genes based on total RNA of mononuclear cells isolated from a blood sample collected from a subject is performed, it is possible to predict the existence risk and therapeutic effect of a cancer cell population. It becomes possible.

The mononuclear cells targeted by the cancer genetic testing method of the present invention are monocytes and / or macrophages.

Monocytes and macrophages have phagocytic ability as described above. In particular, macrophages are phagocytosed by cancer cells that have been damaged by the above-mentioned NK cells or killer T cells and induced apoptosis. Although details are still unclear, it has been suggested that macrophages themselves may damage cancer cells, thereby inducing apoptosis and phagocytosing the cancer cells. In this way, cancer cells that have induced apoptosis and necrosis are eventually phagocytosed by macrophages. Cells can be obtained efficiently.

Furthermore, the expression analysis in the cancer genetic testing method of the present invention is characterized in that it is a real-time polymerase chain reaction (RT-PCR) analysis. Furthermore, the expression analysis in the cancer genetic testing method according to the present invention is a nucleic acid microarray analysis.

In the present invention, a real-time-PCR (RT-PCR) analysis or a nucleic acid microarray analysis is performed for the expression analysis of cancer-related genes. In the real-time-PCR (RT-PCR) analysis, the partial nucleic acid sequence of a cancer-related gene used as a primer or the partial nucleic acid sequence of a cancer-related gene used as a probe in a nucleic acid microarray analysis It can be designed and synthesized accordingly. Therefore, it is possible to perform an accurate test according to the cancer to be tested, and it is possible to perform an expression analysis with high reproducibility and high sensitivity.

The cancer-related genes targeted by the cancer genetic testing method of the present invention include head and neck cancer, pancreatic cancer, thyroid cancer, biliary tract cancer, lung cancer, non-small cell lung cancer, kidney cancer, stomach cancer, liver cancer, colon cancer, rectum Selected from the group consisting of cancer, transitional cell carcinoma, Merkel cell carcinoma, breast cancer, uterine cancer, ovarian cancer, leukemia, esophageal cancer, skin cancer, bladder cancer, prostate cancer, brain tumor, osteosarcoma, and neuroblastoma It is a related gene.

The cancer-related genes targeted by the cancer genetic testing method of the present invention include bcl-2, CA125, CD44, c-kit, c-met, c-myc, COX-2, Cyclin D1, Cytokeratin-7, Cytokeratin-19, Cytokeratin-20, E2F1, E2F3, FGFR2, Gli1, GPC3, hCGbeta, Her2 / Neu, HLF-1a, HnRNP A2 / B1, hTERT, L-myc, MAGEA4, MAGEA4, MAGEMP4 -2, MMP-9, Muc-1, Muc-4, NSE, RCAS1, Survivin, Thyroglobulin, VEGF-A, VEGF-C, AFP, CEA, CGA, EGFR, MAGE A1, MAGE A3 / A6 MUC-7, ProGRP, PSA, SCC, and can be used at least one cancer-associated gene is selected from one of the group consisting of WT1.

In addition, cancer-related genes targeted by the lung cancer genetic testing method of the present invention are bcl-2, CA125, CD44, c-kit, c-met, c-myc, COX-2, Cyclin D1, Cytokeratin-7, Cytokeratin-19, Cytokeratin-20, E2F1, E2F3, FGFR2, Gli1, GPC3, hCGbeta, Her2 / Neu, HLF-1a, HnRNP A2 / B1, hTERT, L-myc, MAGEA4, MAGEA4, MAGEMP4 -2, MMP-9, Muc-1, Muc-4, NSE, RCAS1, Survivin, Thyroglobulin, VEGF-A, VEGF-C, AFP, CEA, CGA, EGFR, MAGE A1, MAGE A3 / A , MUC-7, ProGRP, PSA, SCC, and can be used at least one cancer-associated gene is selected from one of the group consisting of WT1.

Further, cancer-related genes targeted by the lung cancer genetic testing method of the present invention are COX-2, Cyclin D1, Cytokeratin-19, E2F1, E2F3, FGFR2, Gli1, hCGbeta, Her2 / Neu, HLF-1a, HnRNP A2. / B1, MAGE A4, MAGE A12, mdm2, MMP-9, Muc-1, RCAS1, Survivin, Thyroglobulin, VEGF-A, CEA, CGA, and EGFR selected at least one cancer-related Genes can be used.

The cancer-related genes targeted by the breast cancer genetic testing method of the present invention include bcl-2, CA125, CD44, c-kit, c-met, c-myc, COX-2, Cyclin D1, Cytokeratin-7, Cytokeratin-19, Cytokeratin-20, E2F1, E2F3, FGFR2, Gli1, GPC3, hCGbeta, Her2 / Neu, HLF-1a, HnRNP A2 / B1, hTERT, L-myc, MAGEA4, MAGEA4, MAGEMP4 -2, MMP-9, Muc-1, Muc-4, NSE, RCAS1, Survivin, Thyroglobulin, VEGF-A, VEGF-C, AFP, CEA, CGA, EGFR, MAGE A1, MAGE A3 / A , MUC-7, ProGRP, PSA, SCC, and can be used at least one cancer-associated gene is selected from one of the group consisting of WT1.

Further, cancer-related genes targeted by the genetic testing method for breast cancer of the present invention include c-kit, COX-2, Cyclin D1, Cytokeratin-19, Cytokeratin-20, E2F1, E2F3, FGFR2, hCGbeta, Her2 / Neu, At least one cancer-related gene selected from any of the group consisting of HIF-1a, HnRNP A2 / B1, MAGE A4, MAGE A12, mdm2, MMP-9, Muc-1, Thyroglobulin, VEGF-A, and EGFR Can be used.

The cancer-related genes targeted by the gastric cancer genetic testing method of the present invention include bcl-2, CA125, CD44, c-kit, c-met, c-myc, COX-2, Cyclin D1, Cytokeratin-7, Cytokeratin-19, Cytokeratin-20, E2F1, E2F3, FGFR2, Gli1, GPC3, hCGbeta, Her2 / Neu, HLF-1a, HnRNP A2 / B1, hTERT, L-myc, MAGEA4, MAGEA4, MAGEMP4 -2, MMP-9, Muc-1, Muc-4, NSE, RCAS1, Survivin, Thyroglobulin, VEGF-A, VEGF-C, AFP, CEA, CGA, EGFR, MAGE A1, MAGE A3 / A , MUC-7, ProGRP, PSA, SCC, and can be used at least one cancer-associated gene is selected from one of the group consisting of WT1.

Cancer-related genes targeted by the colorectal cancer genetic testing method of the present invention include bcl-2, CA125, CD44, c-kit, c-met, c-myc, COX-2, Cyclin D1, Cytokeratin-7. , Cytokeratin-19, Cytokeratin-20, E2F1, E2F3, FGFR2, Gli1, GPC3, hCGbeta, Her2 / Neu, HIF-1a, HnRNP A2 / B1, hTERT, L-myc, MAGEA12AMAG MMP-2, MMP-9, Muc-1, Muc-4, NSE, RCAS1, Survivin, Thyroglobulin, VEGF-A, VEGF-C, AFP, CEA, CGA, EGFR, MAGE A1, MAGE A3 / 6, MUC-7, ProGRP, PSA, SCC, and can be used at least one cancer-associated gene is selected from one of the group consisting of WT1.

In addition, cancer-related genes targeted by the genetic testing method for liver cancer of the present invention include bcl-2, CA125, CD44, c-kit, c-met, c-myc, COX-2, Cyclin D1, Cytokeratin-7, Cytokeratin-19, Cytokeratin-20, E2F1, E2F3, FGFR2, Gli1, GPC3, hCGbeta, Her2 / Neu, HLF-1a, HnRNP A2 / B1, hTERT, L-myc, MAGEA4, MAGEA4, MAGEMP4 -2, MMP-9, Muc-1, Muc-4, NSE, RCAS1, Survivin, Thyroglobulin, VEGF-A, VEGF-C, AFP, CEA, CGA, EGFR, MAGE A1, MAGE A3 / A , MUC-7, ProGRP, PSA, SCC, and can be used at least one cancer-associated gene is selected from one of the group consisting of WT1.

The cancer-related genes targeted by the cancer genetic testing method of the present invention are 14-3-3-z (ZETA), ADAM12, β-catenin, Bcl-xl, B-myb, C3G, CD44, CDC25A, CDC25B, CDCP-1, CDK2, CDK4, CDK5, CTAP III / NAP-2, Cyclin B1, Cyclin E1, CYP2A6, DNA Polk (Kappa), DPD, DSG3, DYRK2, EphA2, ERBB3, ERBB3, ERBB4, ERBB4 FOXA1 (HNF3alpha), FOXM1, GPR87, hCdc6, hCdt1, HMGA2, Hrad17, Ki-67, KRT5, KRT6A, KRT6B, KRT14, Mesothelin / ERC, MMP-7, MMP-11 Mucin5 (MUC5), Nanog, N-myc, NOTCH3, PAX9, PCNA, PDGF-B, PIK3CA, PKCi (Iota), PRDX1, S100A2, S100A4, SOX4, STAT3, TITF1 / TTF1, TP-1, TS, VEGF , VEGFR-2, VEGFR-3, WNT1, TRF1, TRF2, hTR (TERC), hTEP1, BAGE, CALCA, Evi-1, EEF1A2, GAGE, HBV, HCCR 1, HCCR 2, HCV, HTLV, HPV, NCOA4, At least one cancer-related gene selected from any of the group consisting of PSMA and HSP72 can be used.

Further, cancer-related genes targeted by the lung cancer genetic testing method of the present invention include 14-3-3-z (ZETA), ADAM12, β-catenin, Bcl-xl, B-myb, C3G, CD44, CDC25A, CDC25B, CDCP-1, CDK2, CDK4, CDK5, CTAP III / NAP-2, Cyclin B1, Cyclin E1, CYP2A6, DNA Polk (Kappa), DPD, DSG3, DYRK2, EphA2, ERBB3, ERBB3, ERBB4, ERBB4 FOXA1 (HNF3alpha), FOXM1, GPR87, hCdc6, hCdt1, HMGA2, Hrad17, Ki-67, KRT5, KRT6A, KRT6B, KRT14, Mesothelin / ERC, MMP-7, MMP-1 , Mucin5 (MUC5), Nanog, N-myc, NOTCH3, PAX9, PCNA, PDGF-B, PIK3CA, PKCi (Iota), PRDX1, S100A2, S100A4, SOX4, STAT3, TITF1 / TTF1, TP, TS, VEGF At least one cancer-related gene selected from the group consisting of 1, VEGFR-2, VEGFR-3, WNT1, TRF1, TRF2, hTR (TERC), and hTEP1 can be used.

In addition, the cancer-related genes targeted by the breast cancer genetic testing method of the present invention are FGFR4 (Fibroblast growth factor 4), SERENBP1 (Selenium binding protein 1), POSTN (Periostin), OsteBlastPI, LUC. Serine (or systemine) proteinase inhibitor), HSP-47 (heat shock protein 47), CBP1 (collagen binding protein 1), KDELR3 DEAD (Asp-Glu-Alo-Aply17Ap) (Parathymosin), HIST2H2BE (Histone 2, H2be), TUSC3 (Tumor suppressor candidate 3), ZNF516 (Zinc finger protein 516), BMI1 Polycomb group ring finger 4, ACAD11, INPP4B, ESR1 (Estrogen receptor 1), ACAA2, Myosin VB , CYB5A (Cytochrome b-5), CLIP4 (CAP-GLY domain containing linker, family family, member 4), AYTL1 Hyperthetic protein, RALGPS1 (RALGPS1) al GEF with PH domain and SH3 binding motif 1), PSMB10 [Proteasome (prosome, macropain) subunit, beta type, 10], LRBA (LPS-responsive vesicle trafficking beach and anchor containing), UBE1 (Ubiquitin-activating enzyme E1, FBP1 (Fructose-1,6-bisphosphatase 1), COL6A3 (Collagen, type VI), alpha 3, TM9SF2, Transmembrane 9, superfamily member 2, TAP1 T ansporter 1, ATP-binding cassette, sub-family B (MDR / TAP), DIP2B KIAA1463 protein, TncRNA Trophoblast-derived noncoding RNA, EIF4G1 Eukaryotic translation initiation factor 4 gamma, 1, PRIM2 primase, polypeptide 2A, 58kDa, POR P450 ( cytochrome) oxidoreductase, FANCG Fanconi anemia, complementation group G, EXOSC7 Exosome component 7, FDPS Farnesyl iphosphate synthase (farnesyl pyrophosphate synthetase, dimethylallyltranstransferase, geranyltranstransferase), C5orf4 Chromosome 5 open reading frame 4, CDCA4 Cell division cycle associated 4, H2AFV H2A histone family, member V, FABP3 Fatty acid binding protein 3, muscle and heart (mammary-derived growth inhibitor), CKLF Chemokine-like fact or, MGC4308 Hypothetical protein MGC4308, FADS1 Fatty acid desaturase 1, SFTPB Surfactant, pulmonary-associated protein B, PLA2R1 Phospholipase A2 receptor 1,180kDa, SELENBP1 Selenium binding protein 1, FARSB Phenylalanine-tRNA synthetase-like, beta subunit, MEF2D MADS box transcription enhancer factor 2, polypeptide D (myocycle enhancer facto) 2D), NUTF2 Nuclear transport factor 2, ID3 Inhibitor of DNA binding 3, dominant negative helix-loop-helix protein, ID2 Inhibitor of DNA binding 2, dominant negative helix-loop-helix protein, PRG2 Proteoglycan 2, bone marrow, H2AFX H2A histone family, member X, WSB1 WD repeat and SOCS box-containing 1, TRIM33 Tripartite motif-containing 33, R B5C RAB5C, member RAS oncogene family, PRIM2 primase, polypeptide 2A, 58kDa, RAB21 RAB21, member RAS oncogene family, SSTK Serine / threonine protein kinase SSTK, ST6GAL1 ST6 beta-galactosamide alpha-2,6-sialyltranferase 1, IRF4 Interferon regulatory factor 4. LRBA LPS-responsive vesicle trafficking, beach and anchor containment, IL13RA1 Interleuk in 13 receptor, alpha 1, ESR1 Estrogen receptor 1, MFAP2 Microfibrillar-associated protein 2, STAT1 Signal transducer and activator of transcription 1,91kDa, PPFIA1 Protein tyrosine phosphatase, receptor type, f polypeptide (PTPRF), interacting protein (liprin), alpha 1, MYO1C Myosin IC, PPFIA4 Protein tyrosine phosphate, receptor type, f poly eptide (PTPRF), interacting protein (liprin), alpha 4, RAB31 RAB31, member RAS oncogene family, INPP4B Inositol polyphosphate-4-phosphatase, type II, 105kDa, ESR1 Estrogen receptor 1, and MMP11 from Matrix metalloproteinase 11 (stromelysin 3) At least one cancer-related gene selected from any of the groups can be used.

The cancer-related genes targeted by the gastric cancer genetic testing method of the present invention are TFF1, COX-2, β-Catenin, c-myc, c-jun, APOC1, YF13H12, CDH17, FUS, APOE, S100A11, GRO1. , V-jun, v-raf-1, nibrin, humanin, bcl-2, CAS, semaphorin V, CDK4, CKS1, CKS2, cyclin C, cyclin D, cyclin E, CDC25B, Ki-67, MIA, PCNA, DNA topoisomerase IV-2a, NE-DIg, Retinoblastoma, binding protein-4, IRF7, HOXB7, NFIL3, SRY-box4and 9, IGF-2, TGF-b3, PLAB FGF-2, HGF, HDGF, nm23, C-ERBB2, C-ERBB3, FGFR-4, IGFR-2, EphB2, H1Hisminamine receptor, Hemopoetic cell kinase-1, SKY, GRB-2 and 7, p38in MAPKAx 2, Transport; Solution carrier family-2,16,25, Transferrin receptor, Cytokeratin-1, catenin-b1andg, matrixmetalloproteses; MMP-1, -2, -3, -7, -10, -11 -14, -16, uPA, catepsin-B, e's-K, LI- and OB-cadherin, E-cadher in, collagenes [I, III, IV, V, VI, VII, XVIII] from a3-integrin, fibroblast collagenase inhibitor, S100A4, CD9, IL-8, osteointin, thrombospondin-2, ont, and veston At least one cancer-related gene selected from any of the groups can be used.

Also, cancer-related genes targeted by the colorectal cancer genetic testing method of the present invention are CCSA (Colon cancer-specific antigen) -2, CCSA (Colon cancer-specific antigen) -3, CCSA (Colon cancer-specific antigen). -4, CCSA (Colon cancer-specific antigen) -5, CP (Cancer-placementa) -1, Alpha-catenin, REG1A (Regenerating isletted-private 1 alpha), DPEP1 (Dipeptidase 1) , HERV-H, NLF1 uclear localized factor 1 33.1, FOXQ1 Forkhead box Q1 24.4, MSX2 Msh homeobox homologue 2 22.2, ASCL2 Achaete-scute complex-like 2 17.3, MSX1 Msh homeobox homologue 1 8.5, IRX3 Iroquois homeobox protein3 8.4, GRHL3 Grainyhead-like 3, 7.9, TRIM29 Tripartite motif-continging 29 7.4, ETV4 Ets variant genete4 (E1A enhancer bindingEprote5) 4, ARNTL2 Aryl hydrocarbon receptor nuclear translocator-like 2 5.3, TEAD4 TEA domain family member 4 5.2, SP5 Sp5 transcription factor 5.2, HES6 Hairy and enhancer of split64.6, TBX3 T-box 3 4.6 , NFE2L3 Nuclear factorREG1B Regenerative islet-delivered 1h 75.8, REG3A Regenerative islet-delivered 3a 29.5, TACSTD2 Tumor-associative transducer 2 21.4, IL-8 Interleukin-8 14.7, SERPINB5 Serpin peptidase inhibitor, clade B, member5 (Maspin) 13.8, REG1A Regenerative1. .5, DUSP4 Dual specificity phosphatase 4 7.4, REG4 Regenerating islet-delivered family, member 4.6.8, PHLDA1 Plextrin homology-likeness-like A, member 1 6.0, LCN2 Lipocalin 2 (oncogene 24p3) 5.7, RTEL1 Regulator of telomere elongation helicity 1, 5.6, TGFBI Transforming inc. 4.8, TDGF1 Teratocarcinoma-derived growth factor 1 4.7, TNFRSF6B Tumor necrosis factor superfamily, member 6b, decoy 4.5 1 Deleted in malignant brain tumors 1 4.2, TNFRSF10C Tumor necrosis factor receptor superfamily, member 10c, decoy 4.1, ANGPTL1 Angiopoietin-likeDSG4 Desmoglein 4 5.9, CLDN8 Claudin 8 25.8, CDH19 Cadherin 19, type 2 8 .3, CEACAM7 Carcinoembryonic antigen-related cell adhesion molecule 7.8.3, CLDN23 Claudin 23 8.0, NRXN1 Neuroxin 7.1, PCDH19 rotocadherin 19 6.8, NLGN4X Neuroligin 4, X-linked 6.0, TNXB Tenascin XB 5.6, MUCDHL Mucin and cadherin-like 5.1, PCDH9 ProC1C9C9C9 TFIIIA, IGF-2, TGF-beta, NADH dehydrogenase 2 subunit, Filamin, fibrectin, catepsin H, collagen I a2, collagens (III, IV, V, X), lactadherin 24, HME nectadrin), TIMP-1, a6-integrin, MMP-1, -2-3, -7, -11, -13-10, IL-8, Osteolectin, Thrombopondin-2, VEGF, and Osteopontin At least one cancer-related gene selected from can be used.

In addition, the cancer-related genes targeted by the genetic testing method for liver cancer of the present invention are Fibronincinc X02761; K00799; K02273, Tubulin alpha 1 subunit K00558, Matrix metalloproteinase 14 D26512; repair protein; RAD23A D21235, Ubiquitin-conjugating enzyme E2, M74524, Trafficking protein Neutropil gelatinase-associated lipocalin recursor lipocalin 2 X99133, TRAM protein X63679, ADP / ATP carrier protein J02683, Transcription factor High mobility group protein M23619, Growth factor Insulin-stimulated protein kinase 1 U08316, GTP binding protein Transforming protein rhoA H12 L25080, IFN response Interferon gamma antagonist A25270, Cytokines Macrophage inhibitory cyto ine 1 AF019770, Down-regulated genes in HCCs tumor tissues, Immune system IgG, IgG Kb M63438 + U72063, IgG3; IgG1 L; IgG1 K; IgG1 Fcb D78345 + Y14737, IgA1; IGHAb J00220 + S71043, IgC mu heavy chain constant regionb X57086; X57331, IgG receptor FC large subunit P51 U12255, Lymphocyte antigen M81141, Metabolic pathway Wayne-homocystineine S-methyltran feraseb U50929, Methylenetetrahydrofolate dehydrogenase J04031, Aldehyde oxidase L11005, Uridine diphosphoglucose pyrophosphorylase U27460, Metalloproteinase inhibitor 1 X03124, Metallothionein-IIIb D13365; M93311, Growth factor alpha-2-macroglobulinb M11313, alpha-2-macroglobulin receptor-associated protein M63959, TGF- betac S81439, hepatoc te growth factor-like protein D49742; S83182, NGF-inducible anti-proliferative protein PC3 U72649, Early growth response protein 1b X52541; M62829, Insulin-like growth factor-binding protein 3c M31159; M35878, Insulin-like growth factor binding protein 4 M62403, IFN response Interferon-induced protein MXA M33882, Interferon-inducible prote n 9-27 J04164, Hormones, receptors Nuclear hormone receptor L76571, Extracellular matrix Plasminogenb, c X05199, Haemoglobin alpha subunitb, c V00491, Decorin M14219, Tumour associated c-myc oncogene V00568, Cell cycle cyclin-dependent kinase inhibitor 1 U09579; L25610 , DNA-binding and chromatin protein, DNA-binding protein CPBP U44975, and GTP It is possible to use at least one cancer-associated gene is selected from one of the group consisting of inding protein Transforming protein rhoB X06820.

In addition, cancer-related genes targeted by the genetic testing method for liver cancer related to HBV or HCV according to the present invention are CDC28 protein kinase 2 X54942, CDC27HS protein U00001, Extracellular matrix Ingredient beta 4X57887M6187 J05211, Metabolic pathway Procalagen C proteinase M22488 + U50330, Growth factor Platelet-derived growth factor A subunit X06374, CMP-N-acetylet alactosamide-alpha-2,6-sialytransferase Betaine-homocysteine S-methyl-transferase U50929, Dihydro-orotate dehydrogenaseb M94065, Cytidine deaminase L27943, Aldehyde oxidaseb L11005, Aminoacylase 1 L07548, Methylenetetrahydrofolate dehydrogenaseb J04031, Metallothionein-IIIb X62822, Immune system IgG receptor FC large subunit P51b U12255, M ajor histocompatibiity complex enhancer-binding protein MAD3 M69043, Leukocyte IgG receptor J04162, FC-epsilon-receptor gamma subunit M33195, HLA-DR antigen-associated invariant subunit X00497, Growth factor Hepatocyte growth factor activator D14012, Growth factor receptor-bound protein 2 isoform L29511; M96995, Epidermal growth factor re eptor, oncogene ERBBc X00588; K03193, EGF response factor 1 X79067, IFN response STAT-induced STAT inhibitor 2b AB004903, STAT-induced STAT inhibitor 3b, c, Hormone Insulin-induced protein 1b U96876, Cytokine receptor Interleukin-1 receptor antagonist protein precursor M63099, Apoptosis PIG7; gene induced by p53 AF0101212, Growth arrest and D A-damage-inducible protein M60974, Cell cycle Cyclin-dependent kinase inhibitor 1; P21 U09579; L25610, G protein Guanine nucleotide-binding protein G U31383, Transcription factor Signal transducer and activator of transcription 3b L29277, Polyhomeotic 2 homolog U89278, Liver specific Haemoglobin alpha subunit, c V00491, Regulatory protein Gravin M96322, Binding protein DNAX activation protein 12 AF019562, Unknown function KIAA0022 Gene D14664, Growth factor Transforming growth factor, beta-inducedb, Metabolic pathway Lactate dehydrogenase A X02152, Aldehyde oxidaseb L11005, Nucleoside-diphosphate kinase Y07604, Cytoskeleton Alpha 1 cateninb, c D13866; D14705, Beta cateinc X87 38; Z19054, Collagen 6 alpha 1 subunit X15879, Type II cytoskeletal 8 keratin M34225, Growth factor, receptor Growth factor receptor-bound protein 2 and 3 isoforms L29511; M96995, Hormone Progesteron receptor-associated protein (HSC70) U28918, Tumour associated c -Jun N-terminal kinase 2 L31951, shb proto-oncogene X75342, Transport protein ADP / ATP carrier proteinb J02683, Immune system FC-epsilon-receptor gamma subunitb M33195, Cytokine receptor Interleukin-1 receptor antagonist protein precursor M63099, Regulatory protein Protein kinase C inhibitor 1 U51004, GTP binding protein Rho GTPase activating protein 4b X78817, Guanine nucleotide-binding protein G (l) alpha subunit M17 19, and Genes specifially modulated in HCC developed consisting on cirrhotic tissue can be used at least one cancer-associated gene is selected from any group.

In the present invention, a partial nucleic acid sequence of a cancer-related gene used as a primer in a real-time-PCR (RT-PCR) analysis or a partial nucleic acid sequence of a cancer-related gene used as a probe in a nucleic acid microarray analysis is appropriately selected. Thus, it is possible to predict the existence risk and therapeutic effect of the cancer population in the early stage or prognosis of the cancer.

The cancer genetic testing method of the present invention is based on a significant difference between the expression level of a cancer-related gene obtained from a blood sample of a healthy person and the expression level of a cancer-related gene obtained from a blood sample of a patient. It is characterized by predicting the existence risk and / or therapeutic effect of a cancer cell population.

In the present invention, absolute or relative between the expression level of a cancer-related gene in a healthy person and the expression level of a cancer-related gene in a patient obtained from real-time-PCR (RT-PCR) analysis or nucleic acid microarray analysis Since the determination is based on a significant difference in the expression level, the existence risk of the cancer cell population and / or the therapeutic effect can be predicted easily and with high accuracy.

The cancer genetic testing method of the present invention predicts the existence risk and / or therapeutic effect of a cancer cell population based on the analysis of the expression level of a cancer-related gene that is usually hardly observed in healthy individuals. It is characterized by.

In the present invention, in healthy individuals, the risk of cancer cell populations and / or therapeutic effects are predicted based on the expression level of cancer-related genes that are normally almost unexpressed. It is not necessary to analyze the expression level of the gene, and it is possible to predict the existence risk of the cancer cell population and / or the therapeutic effect quickly and with high accuracy.

According to the cancer genetic testing method of the present invention, it is possible to predict the existence risk of a cancer cell population and the therapeutic effect with high accuracy and early or prognosis.

It is a flowchart for demonstrating the process process of the genetic test method of the cancer concerning embodiment of this invention. It is a figure explaining a cancer related gene. It is a figure explaining a cancer related gene. It is a table | surface showing the result of the expression analysis of the cancer related gene in a lung cancer patient and a healthy subject. It is a table | surface showing the result of the expression analysis of the cancer related gene in a lung cancer patient and a healthy subject. It is a figure explaining a cancer related gene. It is a table | surface showing the result of the expression analysis of the cancer related gene in a breast cancer patient and a healthy subject. It is a table | surface showing the result of the expression analysis of the cancer related gene in a breast cancer patient and a healthy subject. It is the table | surface which put together the evaluation of the genetic test method of the cancer concerning this invention.

Hereinafter, embodiments of the present invention will be described with reference to the drawings. In addition, this invention is not limited to the following description, In the range which does not deviate from the summary of this invention, it can change suitably.

FIG. 1 is a flowchart for explaining processing steps of a cancer genetic testing method according to an embodiment of the present invention. As shown in FIG. 1, in the cancer genetic testing method of this embodiment, a buffy coat layer separation step 10 by centrifugation of a blood sample, a mononuclear cell separation step 20 by density gradient centrifugation, Extraction step 30 of total RNA from nucleated cells, synthesis step 40 of complementary DNA (hereinafter referred to as cDNA) by reverse transcription reaction from total RNA, and real-time PCR (RT-) for cancer-related genes PCR) analysis step 50 and DNA microarray analysis step 60 as a nucleic acid microarray for cancer-related genes.

First, the buffy coat layer separation step 10 by centrifugation of a blood sample is a step of fractionating a buffy coat layer rich in white blood cells by centrifugation from a whole blood sample collected from a subject. For example, when a whole blood sample is centrifuged (2400 × g, 10 minutes) at room temperature, the whole blood sample is separated into three layers. The middle layer fractionated at this time is a buffy coat layer, which is a fraction rich in leukocytes. Then, the buffy coat layer is carefully separated, and then mononuclear cells are separated by density gradient centrifugation.

The mononuclear cell separation step 20 by density gradient centrifugation is a step of separating mononuclear cells from the buffy coat layer obtained by centrifugation of the whole blood sample. The density gradient centrifugation is not particularly limited. For example, sucrose, sedimentation rate method using density gradient by dextran or histopac, Ficoll method, cesium chloride, cesium sulfate, etc. It can be carried out by a method such as isodensity centrifugation using a density gradient with an aqueous solution. In addition, for example, by using a blood separation solution such as Lymphoprep (registered trademark) manufactured by Cosmo Bio, the separation step can be performed in one step. It can be done according to this.

The extraction step 30 of total RNA from mononuclear cells is not particularly limited, but total RNA can be extracted by a known or similar method. For example, when using a commercially available RNA extraction kit, it can carry out according to description of the attached instruction manual.

The cDNA synthesis step 40 by reverse transcription reaction from total RNA is not particularly limited. For example, a method of selectively synthesizing cDNA from the 3 ′ side of mRNA using an oligo dT primer, specific to a known cDNA sequence. CDNA can be synthesized by a method using a gene-specific primer, or by using a random primer, or a known or similar method, such as starting synthesis of cDNA from the entire region of mRNA.

Next, using the synthesized cDNA, a real-time PCR (RT-PCR) analysis step 50 targeting cancer-related genes or a DNA microarray analysis step 60 targeting cancer-related genes is performed.

At this time, the partial nucleic acid sequence in the cancer-related gene of the primer used in the real-time-PCR (RT-PCR) analysis step 50 or the probe used in the DNA microarray analysis step 60 is known in the art or by a method analogous thereto. Can be produced. For example, when the partial nucleic acid sequence to be used is an oligonucleotide, it can be prepared by an organic chemical method, and when it is a partial nucleic acid sequence having a relatively large size, a cell system or a cell-free system It can produce by the biological technique of. The length of the partial nucleic acid sequence is not particularly limited as long as the synthesized cDNA is sufficiently long to allow hybridization, but is at least 15 bp, preferably at least 20 bp, more preferably at least 25 bp, and even more preferably. Is at least 30 bp.

In the real-time-PCR (RT-PCR) analysis step 50, fluorescence is emitted in proportion to the amount of cancer-related gene amplified by the PCR reaction, and the change in fluorescence intensity with time is measured. In this case, the method of emitting fluorescence is not particularly limited. For example, SYBR Green, which is a dye that binds to DNA, is used, or a TaqMan probe that specifically binds to an amplified cancer-related gene is used. A method known in the art or a method analogous thereto can be used. The measured expression level of the cancer-related gene of the patient is compared with the expression level of the cancer-related gene of the healthy person, and determined with an absolute or relative significant difference. Based on these determination results Presence risk of cancer cell population and / or therapeutic effect is predicted. The measured expression level of the cancer-related gene of the patient may be collated with the expression level data of the cancer-related gene accumulated in advance.

In the DNA microarray analysis step 60, 1) synthesis of double-stranded DNA based on the cDNA synthesized in the cDNA synthesis step 40 by reverse transcription reaction from total RNA, and 2) the obtained double-stranded DNA RNA amplification by in vitro transcription reaction based on 3) RNA labeling of healthy person-derived RNA and patient-derived RNA, 4) Cancer-related genes using both fluorescently labeled RNAs as probes 5) The expression change of the cancer-related gene of the patient relative to a healthy person is determined from the fluorescence intensity of the fluorescently labeled RNA hybridized with the probe. And the existence risk of a cancer cell population and / or a therapeutic effect are estimated based on these determination results.

In addition, as a form of the DNA microarray used in the present embodiment, a form known in the art or an equivalent form thereof can be used. For example, an array in which a partial diffusion sequence of a cancer-related gene is directly synthesized on a support. (Affymetrix method), an array in which a partial nucleic acid sequence of a cancer-related gene is immobilized on a support (Stanford method), or the like can be used.

Specifically, in the Affymetrix method, nucleic acid molecules are silicon-based while repeating the process of covering and exposing the silicon substrate as a support with a light-shielding plate called a mask by photolithography technology and solid-phase nucleic acid synthesis technology. An array is made by extending one base at a time. According to this method, since the synthesized partial nucleic acid sequence can be fixed perpendicularly to the support, there are advantages such as high hybridization efficiency and excellent quantification and reproducibility. On the other hand, in the Stanford method, an array is prepared by spotting cDNA prepared in advance or synthetic oligonucleotide directly on a support. According to this method, compared to the Affymetrix method, there is an advantage that an arbitrary partial nucleic acid sequence can be mounted, the running cost is low, and the self-made is easy. Have disadvantages in that cross-hybridization occurs and partial nucleic acid sequences are easily detached from the support. Arrays based on these methods can be appropriately selected in consideration of the number of partial nucleic acid sequences mounted on cancer-related genes, running costs, and the like.

[Example]
Examples of the present invention will be specifically described, but the present invention is not limited to these Examples, and can be appropriately changed without departing from the gist of the present invention.

[Preparation of blood sample]
Based on the results of the PET-CT examination, blood samples were prepared from 50 healthy human subjects, 28 lung cancer patients, and 11 breast cancer patients. It should be noted that the subject is fully explained about the test according to the present invention and has obtained consent to use a blood sample.

[Separation of mononuclear cells]
First, a puffy coat layer rich in leukocytes was obtained from a blood sample collected from a subject by centrifugation. Then, the puffy coat layer was suspended, and gently layered on an equal amount of histopack solution (manufactured by Sigma), followed by density gradient centrifugation to collect mononuclear cells.

[Extraction of total RNA from mononuclear cells]
Total RNA was extracted from the obtained mononuclear cells using Easy-spin ™ Total RNA Extraction Kit (manufactured by iNtron) according to the description in the attached instruction manual.

[Reverse Transcription Reaction and Real-Time-Preparation of 384 Low Density Array for PCR]
From the total RNA obtained, reverse transcription was performed using High Capacity RNA-to-cDNA Master Mix (manufactured by Applied Biosystems) according to the description of the attached instruction manual, and cDNA was synthesized from the total RNA. In addition, 47 cancer-related genes (transcription factors, receptors, angiogenesis-related genes, etc.) shown in FIG. 2 were selected, and a 384 Low density array (manufactured by Applied Biosystems) for real-time PCR was outsourced.

[Real-time-PCR reaction and statistical processing]
For each synthesized cDNA, a reaction solution was prepared using Master Mix for real-time PCR (Applied Biosystems) according to the description in the attached instruction manual. Then, the prepared reaction solution is subjected to an externally prepared 384 Low density array, and after a real-time-PCR reaction using a 7900HT real-time-PCR apparatus (Applied Biosystems), each specimen is expressed with respect to the expression ratio of each cancer-related gene. Statistical processing was performed. As an endogenous control, the expression ratio of each cancer-related gene to the GAPDH gene was calculated.

[Analysis result]
In healthy subjects, lung cancer patients, and breast cancer patients, the expression ratio of cancer-related genes to the endogenous control gene (GAPDH gene) is calculated using RQ software attached to the 7900HT real-time PCR apparatus (Applied Biosystems), For each specimen, statistical processing was performed on the expression ratio of each cancer-related gene. Specifically, with respect to 50 healthy subjects, the average value + 2SD is set as the reference range, and those exceeding the reference range are defined as cancer risks, and the boxes shown in the tables shown in FIGS. 4, 5, 7, and 8 Expressed by painting.

First, we examined the risk assessment of lung cancer. 4 and 5 are tables showing the results of the expression analysis of cancer-related genes in lung cancer patients and healthy individuals for each cancer-related gene extracted from the 22 cancer-related genes shown in FIG. As shown in FIG. 4 and FIG. 5, the proportion of subjects exceeding the reference range of one or more cancer-related genes among the extracted cancer-related genes is about 20% for healthy subjects and about about 20% for lung cancer patients. 90%.

Next, we examined risk assessment for breast cancer. FIG. 7 and FIG. 8 are tables showing the results of the expression analysis of cancer-related genes in breast cancer patients and healthy individuals for each cancer-related gene extracted from the 19 cancer-related genes shown in FIG. As shown in FIG. 7 and FIG. 8, the proportion of subjects who exceeded the reference range of one or more cancer-related genes among the extracted cancer-related genes was 22% for healthy subjects, and about 20% for breast cancer patients. 82%.

FIG. 9 is a table summarizing the evaluation of the cancer genetic testing method according to the present invention for lung cancer or breast cancer. As shown in FIG. 9, according to the cancer genetic testing method of the present invention, the specificity was 80% and the sensitivity was about 90% in the detection of lung cancer. Further, in the discrimination of breast cancer, the specificity was 78% and the sensitivity was about 82% as the detection rate.

Based on these facts, attention is paid to genes included in the cancer-related gene group shown in FIG. 3 for lung cancer, and genes included in the cancer-related gene group shown in FIG. 6 for breast cancer. The used genetic testing method for cancer according to the present invention is considered useful as a testing method for lung cancer or breast cancer.

In the description of this example, 22 lung cancer-related genes were extracted for lung cancer risk assessment, and 19 breast cancer-related genes were extracted for breast cancer risk assessment. The gene group applied to is not limited to this.

For example, regarding lung cancer risk assessment, in addition to the aforementioned lung cancer-related genes, 14-3-3-z (ZETA), ADAM12, β-catenin, Bcl-xl, B-myb, C3G, CD44, CDC25A, CDC25B , CDCP-1, CDK2, CDK4, CDK5, CTAP III / NAP-2, Cyclin B1, Cyclin E1, CYP2A6, DNA Polk (Kappa), DPD, DSG3, DYRK2, EphA2, ERBB3, ERBB4, EGF2, AGF1A (HNF3alpha), FOXM1, GPR87, hCdc6, hCdt1, HMGA2, Hrad17, Ki-67, KRT5, KRT6A, KRT6B, KRT14, Mesothelin / ERC, MMP-7, MM -11, Mucin5 (MUC5), Nanog, N-myc, NOTCH3, PAX9, PCNA, PDGF-B, PIK3CA, PKCi (Iota), PRDX1, S100A2, S100A4, SOX4, STAT3, TITF1 / TTF1, TP, TS, At least one cancer-related gene selected from any of the group consisting of VEGFR-1, VEGFR-2, VEGFR-3, WNT1, TRF1, TRF2, hTR (TERC), and hTEP1 can be used.

Regarding breast cancer risk assessment, in addition to the above-mentioned breast cancer-related genes, FGFR4 (Fibroblast growth factor receptor 4), SERENBP1 (Selenium binding protein 1), POSTN (PeriostinClinsePlN, FastBlastClinPlSlP) (Or systemine) proteinase inhibitor), HSP-47 (heat shock protein 47), CBP1 (collagen binding protein 1), KDELR3 DEAD (Asp-Glu-Ala-Adipolopopp) p MS (Parathymosin), HIST2H2BE (Histone 2, H2be), TUSC3 (Tumor suppressor candidate 3), ZNF516 (Zinc finger protein 516), BMI1 Polycomb group ring finger 4, ACAD11, INPP4B, ESR1 (Estrogen receptor 1), ACAA2, Myosin VB, CYB5A (Cytochrome b-5), CLIP4 (CAP-GLY domain containing linker family family, member 4), AYTL1 Hyperthetic protein, RALGPS1 (Ral GEF with PH domain and SH3 binding motif 1), PSMB10 [Proteasome (prosome, macropain) subunit, beta type, 10], LRBA (LPS-responsive vesicle trafficking beach and anchor containing), UBE1 (Ubiquitin-activating enzyme E1, FBP1 (Fructose-1,6-bisphosphatase 1), COL6A3 (Collagen, type VI), alpha 3, TM9SF2 Transmembrane 9 superfamily member 2, TAP1 Transporter 1, ATP-binding cassette, sub-family B (MDR / TAP), DIP2B KIAA1463 protein, TncRNA Trophoblast-derived noncoding RNA, EIF4G1 Eukaryotic translation initiation factor 4 gamma, 1, PRIM2 primase, polypeptide 2A, 58kDa, POR P450 ( cytochrome) oxidoreductase, FANCG Fanconi anemia, complementation group G, EXOSC7 Exosome component 7, FDPS Farnesy diphosphate synthase (farnesyl pyrophosphate synthetase, dimethylallyltranstransferase, geranyltranstransferase), C5orf4 Chromosome 5 open reading frame 4, CDCA4 Cell division cycle associated 4, H2AFV H2A histone family, member V, FABP3 Fatty acid binding protein 3, muscle and heart (mammary-derived growth inhibitor), CKLF Chemokine-like fa ctor, MGC4308 Hypothetical protein MGC4308, FADS1 Fatty acid desaturase 1, SFTPB Surfactant, pulmonary-associated protein B, PLA2R1 Phospholipase A2 receptor 1,180kDa, SELENBP1 Selenium binding protein 1, FARSB Phenylalanine-tRNA synthetase-like, beta subunit, MEF2D MADS box transcription enhancer factor 2, polypeptide D (myocycle enhancer fac) or 2D), NUTF2 Nuclear transport factor 2, ID3 Inhibitor of DNA binding 3, dominant negative helix-loop-helix protein, ID2 Inhibitor of DNA binding 2, dominant negative helix-loop-helix protein, PRG2 Proteoglycan 2, bone marrow, H2AFX H2A histone family, member X, WSB1 WD repeat and SOCS box-containing 1, TRIM33 Tripartite motif-containing 33 RAB5C RAB5C, member RAS oncogene family, PRIM2 primase, polypeptide 2A, 58kDa, RAB21 RAB21, member RAS oncogene family, SSTK Serine / threonine protein kinase SSTK, ST6GAL1 ST6 beta-galactosamide alpha-2,6-sialyltranferase 1, IRF4 Interferon regulatory factor 4. LRBA LPS-responsive vesicle trafficking, beach and anchor containment, IL13RA1 Interle ukin 13 receptor, alpha 1, ESR1 Estrogen receptor 1, MFAP2 Microfibrillar-associated protein 2, STAT1 Signal transducer and activator of transcription 1,91kDa, PPFIA1 Protein tyrosine phosphatase, receptor type, f polypeptide (PTPRF), interacting protein (liprin), alpha 1, MYO1C Myosin IC, PPFIA4 Protein tyrosine phosphate, receptor type, f po ypeptide (PTPRF), interacting protein (liprin), alpha 4, RAB31 RAB31, member RAS oncogene family, INPP4B Inositol polyphosphate-4-phosphatase, type II, 105kDa, ESR1 Estrogen receptor 1, and MMP11 from Matrix metalloproteinase 11 (stromelysin 3) At least one cancer-related gene selected from any of the groups can be used.

Regarding gastric cancer risk assessment, bcl-2, CA125, CD44, c-kit, c-met, c-myc, COX-2, Cyclin D1, Cytokeratin-7, Cytokeratin-19, Cytokeratin-20, E2F1, E2F3, FGFR2, Gli1, GPC3, hCGbeta, Her2 / Neu, HIF-1a, HnRNP A2 / B1, hTERT, L-myc, MAGE A4, MAGE A12, mdm2, MDR1, MMP-2, MMP-9, Muc-1 , Muc-4, NSE, RCAS1, Survivin, Thyroglobulin, VEGF-A, VEGF-C, AFP, CEA, CGA, EGFR, MAGE A1, MAGE A3 / A6, MUC-7, ProGRP In addition to PSA, SCC, and WT1, TFF1, COX-2, β-Catenin, c-myc, c-jun, APOC1, YF13H12, CDH17, FUS, APOE, S100A11, GRO1, v-jun, v-raf- 1, nibrin, humanin, bcl-2, CAS, semaphorin V, CDK4, CKS1, CKS2, cyclin C, cyclin D, cyclin E, CDC25B, Ki-67, MIA, PCNA, DNA topoisomerase EbemoRaseBra-2N , Binding protein-4, IRF7, HOXB7, NFIL3, SRY-box4and 9, IGF-2, TGF-b3, PLAB, FGF-2, HGF, HD F, nm23, C-ERBB2, C-ERBB3, FGFR-4, IGFR-2, EphB2, H1Histamine receptor, Hemopoetic cell kinase-1, SKY, GRB-2and 7, p38 MAPK, Axin-2Transport; family-2, 16, 25, Transferrin receptor, Cytokeratin-1, catenin-b1and g, matrix metalloproteases; MMP-1, -2, -3, -7, -10, -11, -12, -14, -16, uPA, catepsin-B, e's-K, LI- and OB-cadherin, E-cadherin, collages I, III, IV, V, VI, VII, XVIII] from a3-integrin, fibroblast collagenase inhibitor, S100A4, CD9, IL-8, Ostectin, Thrombopondin-2, VEGF, and Ostepondinteinbondinte At least one cancer-related gene selected from can be used.

Regarding the risk assessment of colorectal cancer, bcl-2, CA125, CD44, c-kit, c-met, c-myc, COX-2, Cyclin D1, Cytokeratin-7, Cytokeratin-19, Cytokeratin-20, E2F1 , E2F3, FGFR2, Gli1, GPC3, hCGbeta, Her2 / Neu, HIF-1a, HnRNP A2 / B1, hTERT, L-myc, MAGE A4, MAGE A12, mdm2, MDR-1, MMP-2, MMP-9, Muc-9 1, Muc-4, NSE, RCAS1, Survivin, Thyroglobulin, VEGF-A, VEGF-C, AFP, CEA, CGA, EGFR, MAGE A1, MAGE A3 / A6, MUC-7, ProGR , PSA, SCC, and WT1, as well as CCSA (Colon cancer-specific antigen) -2, CCSA (Colon cancer-specific antigen) -3, CCSA (Colon cancer-specific antigen) -4, CCSAcicicol-spec (on-line) antigen-5, CP (Cancer-placementa) -1, Alpha-catenin, REG1A (Regenerating islet-derived 1 alpha), DPEP1 (Dipeptidase 1), PAP (pancreatity-AssociatedHN) zed factor 1 33.1, FOXQ1 Forkhead box Q1 24.4, MSX2 Msh homeobox homologue 2 22.2, ASCL2 Achaete-scute complex-like 2 17.3, MSX1 Msh homeobox homologue 1 8.5, IRX3 Iroquois homeobox protein3 8 .4, GRHL3 Grainyhead-like 3 7.9, TRIM29 Tripartite motif-continging 29 7.4, ETV4 Ets variant genent4, E1A enhancer binding4ARL4E1 hydrocarbon receptor nuclear translocator-like 2 5.3, TEAD4 TEA domain family member 4 5.2, SP5 Sp5 transcription factor 5.2, HES6 Hairy and enhancer of split64.6, TBX3 T-box 3 4.6, NFE2L3 Nuclear factorREG1B Regenerating islet-delivered 1h 75.8, REG3A Regenerative islet-delivered 3a 29.5, TACSTD2 Tumor-associated calcium signal transducer 2 21.4, IL-8 Interleukin-8 14.7, SERPINB5 Serpin peptidase inhibitor, clade B, member 5 (Maspin) 13.8, REG1A Regenerating islet-delimined 7.5-in. , DUSP4 Dual specificity phosphatase 4.7.4, REG4 Regenerating islet-delivered family, member 4.6.8, PHLDA1 Plextrin homology 1-like. , LCN2 Lipocalin 2 (oncogene 24p3) 5.7, RTEL1 Regulator of telomere elongation helicase 1 5.6, TGFBI Transforming growth factor, h induced 5.2, IGFBP2 Insulin-like growth factor binding protein 2 4.8, TDGF1 Teratocarcinoma- Derived growth factor 1 4.7, TNFRSF6B Tumor necrosis factor receptor superfamily, member 6b, decoy 4.5, DMBT1 Deleted in alignant brain tumors 1 4.2, TNFRSF10C Tumor necrosis factor receptor superfamily, member 10c, decoy 4.1, ANGPTL1 Angiopoietin-likeDSG4 Desmoglein 4 5.9, CLDN8 Claudin 8 25.8, CDH19 Cadherin 19, type 2 8.3, CEACAM7 Carcinoembryonic anti-related cell adhesion molecular 7.8.3, CLDN23 Claudin 23 8.0, NRXN1 Neuroxin 7.1, PCDH19 Protocadherin 19 6.8, NLGN4X Neuroligin 4, X-linked 6.0, TNXB Tenascin XB 5.6, MUCDHL Mucin and cadherin-like 5.1, PCDH9 Protocadherin-9L9C4G9 , IGF-2, TGF-beta, NADH dehydrogenase 2, subunit, Filamin, fibrectin, catthepsin H, collagen I a2, collagens (III, IV, V, X), lactadherin, HME, CD24. nectadrin), TIMP-1, a6-integrin, MMP-1, -2-3, -7, -11, -13-10, IL-8, Osteolectin, Thrombopondin-2, VEGF, and Osteopontin At least one cancer-related gene selected from can be used.

Regarding the risk assessment of liver cancer, bcl-2, CA125, CD44, c-kit, c-met, c-myc, COX-2, Cyclin D1, Cytokeratin-7, Cytokeratin-19, Cytokeratin-20, E2F1, E2F3, FGFR2, Gli1, GPC3, hCGbeta, Her2 / Neu, HIF-1a, HnRNP A2 / B1, hTERT, L-myc, MAGE A4, MAGE A12, mdm2, MDR1, MMP-2, MMP-9, Muc-1 , Muc-4, NSE, RCAS1, Survivin, Thyroglobulin, VEGF-A, VEGF-C, AFP, CEA, CGA, EGFR, MAGE A1, MAGE A3 / A6, MUC-7, ProGRP PSA, SCC, and in addition to WT1, Fibronectinc X02761; K00799; K02273, Tubulin alpha 1 subunit K00558, Matrix metalloproteinase 14 D26512; X83535, Osteonectin; SPARC J03040, DNA damage repair protein UV excision repair protein; RAD23A D21235, Ubiquitin-conjugating enzyme E2, M74524, Trafficking protein, Neutropil gelatinase-associated lipocalin precursor calin 2 X99133, TRAM protein X63679, ADP / ATP carrier protein J02683, Transcription factor High mobility group protein M23619, Growth factor Insulin-stimulated protein kinase 1 U08316, GTP binding protein Transforming protein rhoA H12 L25080, IFN response Interferon gamma antagonist A25270, Cytokines Macrophage inhibitory cytokine 1 AF0197 0, Down-regulated genes in HCCs tumor tissues, Immune system IgG, IgG Kb M63438 + U72063, IgG3; IgG1 L; IgG1 K; IgG1 Fcb D78345 + Y14737, IgA1; IGHAb J00220 + S71043, IgC mu heavy chain constant regionb X57086; X57331, IgG receptor FC large subunit P51 U12255, Lymphocyte antigen M81141, Metabolic pathway Wayne-homosineine S-methyltransferase U5092 9, Methylenetetrahydrofolate dehydrogenase J04031, Aldehyde oxidase L11005, Uridine diphosphoglucose pyrophosphorylase U27460, Metalloproteinase inhibitor 1 X03124, Metallothionein-IIIb D13365; M93311, Growth factor alpha-2-macroglobulinb M11313, alpha-2-macroglobulin receptor-associated protein M63959, TGF-betac S81439, hepatocyte growth fac tor-like protein D49742; S83182, NGF-inducible anti-proliferative protein PC3 U72649, Early growth response protein 1b X52541; M62829, Insulin-like growth factor-binding protein 3c M31159; M35878, Insulin-like growth factor binding protein 4 M62403, IFN response Interferon-induced protein MXA M33882, Interferon-inductive protein 9-27 J0416 , Hormones, receptors Nuclear hormone receptor L76571, Extracellular matrix Plasminogenb, c X05199, Haemoglobin alpha subunitb, c V00491, Decorin M14219, Tumour associated c-myc oncogene V00568, Cell cycle cyclin-dependent kinase inhibitor 1 U09579; L25610, DNA-binding and chromatin protein DNA-binding protein CPBP U44975, and GTP binding protein nTransforming protein rhoB X06820 can be used for at least one cancer-related gene selected from the group consisting of nTransforming protein rhoB X06820.

Furthermore, HBV, or for liver cancer risk assessment associated with HCV, CDC28 protein kinase 2 X54942, CDC27HS protein U00001, Extracellular matrix Integrin beta 4 X53587; X52186, Desmoplakin I & II M77830; J05211, Metabolic pathway Procallagen C proteinase M22488 + U50330, Growth factor Platelet-derived growth factor A subunit X06374, CMP-N-acetylneuraminate-beta-galactosamide alpha-2,6-sialytransferase Betaine-homocysteine S-methyl-transferase U50929, Dihydro-orotate dehydrogenaseb M94065, Cytidine deaminase L27943, Aldehyde oxidaseb L11005, Aminoacylase 1 L07548, Methylenetetrahydrofolate dehydrogenaseb J04031, Metallothionein-IIIb X62822, Immune system IgG receptor FC large subunit P51b U12255, Major histocom patibiity complex enhancer-binding protein MAD3 M69043, Leukocyte IgG receptor J04162, FC-epsilon-receptor gamma subunit M33195, HLA-DR antigen-associated invariant subunit X00497, Growth factor Hepatocyte growth factor activator D14012, Growth factor receptor-bound protein 2 isoform L29511 M96995, Epidermal growth factor receptor, oncage e ERBBc X00588; K03193, EGF response factor 1 X79067, IFN response STAT-induced STAT inhibitor 2b AB004903, STAT-induced STAT inhibitor 3b, c, Hormone Insulin-induced protein 1b U96876, Cytokine receptor Interleukin-1 receptor antagonist protein precursor M63099, Apoptosis PIG7; gene induced by p53 AF0101212, Growth arrest and DNA-damage-ind cible protein M60974, Cell cycle Cyclin-dependent kinase inhibitor 1; P21 U09579; L25610, G protein Guanine nucleotide-binding protein G U31383, Transcription factor Signal transducer and activator of transcription 3b L29277, Polyhomeotic 2 homolog U89278, Liver specific Haemoglobin alpha subunitb, c V00491, Regulatory protein Gravin M96322 , Binding protein DNAX activation protein 12 AF019562, Unknown function KIAA0022 Gene D14664, Growth factor Transforming growth factor, beta-inducedb, Metabolic pathway Lactate dehydrogenase A X02152, Aldehyde oxidaseb L11005, Nucleoside-diphosphate kinase Y07604, Cytoskeleton Alpha 1 cateninb, c D13866; D14705, Beta catininc X87838; Z19054, Co lagen 6 alpha 1 subunit X15879, Type II cytoskeletal 8 keratin M34225, Growth factor, receptor Growth factor receptor-bound protein 2 and 3 isoforms L29511; M96995, Hormone Progesteron receptor-associated protein (HSC70) U28918, Tumour associated c-jun N- terminal kinase 2 L31951, shb proto-oncogene X75342, transport protein ADP / ATP carrier p roteinb J02683, Immune system FC-epsilon-receptor gamma subunitb M33195, Cytokine receptor Interleukin-1 receptor antagonist protein precursor M63099, Regulatory protein Protein kinase C inhibitor 1 U51004, GTP binding protein Rho GTPase activating protein 4b X78817, Guanine nucleotide-binding protein G (L) alpha subunit M17219 and Genes It is possible to use at least one cancer-associated genes selected from ecifially modulated in HCC developed any of the group consisting of on cirrhotic tissue.

In the cancer genetic testing method according to the present invention, the cancer-related genes described above may be used, and cancer risk assessment may be performed based on the expression profile of a single cancer-related gene. Cancer risk assessment may be performed based on the expression profiles of cancer-related genes as a group.

As described above, according to the embodiment of the present invention, it is possible to accurately predict the existence risk of a cancer cell population and the therapeutic effect at an early stage or prognosis.

10 Separation process of buffy coat layer by centrifugation of blood sample 20 Separation process of mononuclear cells by density gradient centrifugation 30 Extraction process of total RNA from mononuclear cells 40 Complementary DNA by reverse transcription reaction from total RNA 50 Real-time PCR (RT-PCR) analysis process for cancer-related genes 60 DNA microarray analysis process for cancer-related genes

Claims (22)

1. Separating mononuclear cells from a blood sample collected from a subject;
   Extracting total RNA from the isolated mononuclear cells;
   Synthesizing complementary DNA from the extracted total RNA;
   A method for examining cancer genes, comprising the step of performing expression analysis on a cancer-related gene in the synthesized complementary DNA.
2. 2. The cancer genetic testing method according to claim 1, wherein the mononuclear cells are monocytes and / or macrophages.
3. 3. The cancer genetic testing method according to claim 1 or 2, wherein the expression analysis for the cancer-related gene is a real-time polymerase chain reaction (RT-PCR) analysis.
4. 3. The cancer genetic testing method according to claim 1 or 2, wherein the expression analysis for the cancer-related gene is nucleic acid microarray analysis.
5. The cancer-related genes are: head and neck cancer, pancreatic cancer, thyroid cancer, biliary tract cancer, lung cancer, non-small cell lung cancer, kidney cancer, stomach cancer, liver cancer, colon cancer, rectal cancer, transitional cell cancer, Merkel cell cancer, breast cancer, uterus A related gene selected from the group consisting of cancer, ovarian cancer, leukemia, esophageal cancer, skin cancer, bladder cancer, prostate cancer, brain tumor, osteosarcoma, and neuroblastoma. The genetic test method for cancer according to 3 or 4.
6. The cancer-related genes are bcl-2, CA125, CD44, c-kit, c-met, c-myc, COX-2, Cyclin D1, Cytokeratin-7, Cytokeratin-19, Cytokeratin-20, E2F1, E2F3, FGFR2. , Gli1, GPC3, hCGbeta, Her2 / Neu, HIF-1a, HnRNP A2 / B1, hTERT, L-myc, MAGE A4, MAGE A12, mdm2, MDR1, MMP-2, MMP-9, Muc-1, Muc- 4, NSE, RCAS1, Survivin, Thyroglobulin, VEGF-A, VEGF-C, AFP, CEA, CGA, EGFR, MAGE A1, MAGE A3 / A6, MUC-7, ProGRP, PSA, SC , And genetic testing method for cancer according to claim 5, wherein the at least one cancer-associated genes selected from any of the group consisting of WT1.
7. The cancer-related genes are bcl-2, CA125, CD44, c-kit, c-met, c-myc, COX-2, Cyclin D1, Cytokeratin-7, Cytokeratin-19, Cytokeratin-20, E2F1, E2F3, FGFR2. , Gli1, GPC3, hCGbeta, Her2 / Neu, HIF-1a, HnRNP A2 / B1, hTERT, L-myc, MAGE A4, MAGE A12, mdm2, MDR1, MMP-2, MMP-9, Muc-1, Muc- 4, NSE, RCAS1, Survivin, Thyroglobulin, VEGF-A, VEGF-C, AFP, CEA, CGA, EGFR, MAGE A1, MAGE A3 / A6, MUC-7, ProGRP, PSA, SC , And genetic testing method of claim 5 wherein the lung, characterized in that at least one cancer-associated genes selected from any of the group consisting of WT1.
8. A method for genetic testing of lung cancer according to claim 7,
   The cancer-related genes are COX-2, Cyclin D1, Cytokeratin-19, E2F1, E2F3, FGFR2, Gli1, hCGbeta, Her2 / Neu, HIF-1a, HnRNP A2 / B1, MAGE A4, MAGE A12Md, MP2 9. A method for genetic testing of lung cancer, comprising at least one cancer-related gene selected from the group consisting of 9, Muc-1, RCAS1, Survivin, Thyroglobulin, VEGF-A, CEA, CGA, and EGFR .
9. The cancer-related genes are bcl-2, CA125, CD44, c-kit, c-met, c-myc, COX-2, Cyclin D1, Cytokeratin-7, Cytokeratin-19, Cytokeratin-20, E2F1, E2F3, FGFR2. , Gli1, GPC3, hCGbeta, Her2 / Neu, HIF-1a, HnRNP A2 / B1, hTERT, L-myc, MAGE A4, MAGE A12, mdm2, MDR1, MMP-2, MMP-9, Muc-1, Muc- 4, NSE, RCAS1, Survivin, Thyroglobulin, VEGF-A, VEGF-C, AFP, CEA, CGA, EGFR, MAGE A1, MAGE A3 / A6, MUC-7, ProGRP, PSA, SC , And breast method genetic testing according to claim 5, wherein the at least one cancer-associated genes selected from any of the group consisting of WT1.
10. A method for genetic testing of breast cancer according to claim 9,
    The cancer-related genes are c-kit, COX-2, Cyclin D1, Cytokeratin-19, Cytokeratin-20, E2F1, E2F3, FGFR2, hCGbeta, Her2 / Neu, HLF-1a, HnRNP A2 / B1, MAGE A4, MAGE A4 A genetic test method for breast cancer, which is at least one cancer-related gene selected from the group consisting of A12, mdm2, MMP-9, Muc-1, Thyroglobulin, VEGF-A, and EGFR.
11. The cancer-related genes are bcl-2, CA125, CD44, c-kit, c-met, c-myc, COX-2, Cyclin D1, Cytokeratin-7, Cytokeratin-19, Cytokeratin-20, E2F1, E2F3, FGFR2. , Gli1, GPC3, hCGbeta, Her2 / Neu, HIF-1a, HnRNP A2 / B1, hTERT, L-myc, MAGE A4, MAGE A12, mdm2, MDR1, MMP-2, MMP-9, Muc-1, Muc- 4, NSE, RCAS1, Survivin, Thyroglobulin, VEGF-A, VEGF-C, AFP, CEA, CGA, EGFR, MAGE A1, MAGE A3 / A6, MUC-7, ProGRP, PSA, SC , And genetic testing method of gastric cancer according to claim 5, wherein the at least one cancer-associated genes selected from any of the group consisting of WT1.
12. The cancer-related genes are bcl-2, CA125, CD44, c-kit, c-met, c-myc, COX-2, Cyclin D1, Cytokeratin-7, Cytokeratin-19, Cytokeratin-20, E2F1, E2F3, FGFR2. , Gli1, GPC3, hCGbeta, Her2 / Neu, HIF-1a, HnRNP A2 / B1, hTERT, L-myc, MAGE A4, MAGE A12, mdm2, MDR1, MMP-2, MMP-9, Muc-1, Muc- 4, NSE, RCAS1, Survivin, Thyroglobulin, VEGF-A, VEGF-C, AFP, CEA, CGA, EGFR, MAGE A1, MAGE A3 / A6, MUC-7, ProGRP, PSA, SC , And genetic testing method for colon cancer according to claim 5, wherein the at least one cancer-associated genes selected from any of the group consisting of WT1.
13. The cancer-related genes are bcl-2, CA125, CD44, c-kit, c-met, c-myc, COX-2, Cyclin D1, Cytokeratin-7, Cytokeratin-19, Cytokeratin-20, E2F1, E2F3, FGFR2. , Gli1, GPC3, hCGbeta, Her2 / Neu, HIF-1a, HnRNP A2 / B1, hTERT, L-myc, MAGE A4, MAGE A12, mdm2, MDR1, MMP-2, MMP-9, Muc-1, Muc- 4, NSE, RCAS1, Survivin, Thyroglobulin, VEGF-A, VEGF-C, AFP, CEA, CGA, EGFR, MAGE A1, MAGE A3 / A6, MUC-7, ProGRP, PSA, SC , And genetic testing method of liver cancer according to claim 5, wherein the at least one cancer-associated genes selected from any of the group consisting of WT1.
14. The cancer-related genes are 14-3-3-z (ZETA), ADAM12, β-catenin, Bcl-xl, B-myb, C3G, CD44, CDC25A, CDC25B, CDCP-1, CDK2, CDK4, CDK5, CTAP III / NAP-2, Cyclin B1, Cyclin E1, CYP2A6, DNA Pol k (Kappa), DPD, DSG3, DYRK2, EphA2, ERBB3, ERBB4, EGF2, EGF9, FOXA1, HOX3alphaMh, CPR2A6 HMGA2, Hrad17, Ki-67, KRT5, KRT6A, KRT6B, KRT14, Mesothelin / ERC, MMP-7, MMP-11, Mucin5 (MUC5), Nanog, -Myc, NOTCH3, PAX9, PCNA, PDGF-B, PIK3CA, PKCi (Iota), PRDX1, S100A2, S100A4, SOX4, STAT3, TITF1 / TTF1, TP, TS, VEGFR-1, VEGFR-2, VEGFR-3 , WNT1, TRF1, TRF2, hTR (TERC), hTEP1, BAGE, CALCA, Evi-1, EEF1A2, GAGE, HBV, HCCR 1, HCCR 2, HCV, HTLV, HPV, NCOA 4, PSMA, and HSP72 6. The cancer genetic testing method according to claim 5, wherein the gene is at least one cancer-related gene selected from any one of the above.
15. The cancer genetic testing method according to claim 14,
    The cancer-related genes are 14-3-3-z (ZETA), ADAM12, β-catenin, Bcl-xl, B-myb, C3G, CD44, CDC25A, CDC25B, CDCP-1, CDK2, CDK4, CDK5, CTAP III / NAP-2, Cyclin B1, Cyclin E1, CYP2A6, DNA Polk (Kappa), DPD, DSG3, DYRK2, EphA2, ERBB3, ERBB4, EGF2, EGF9, FOXA1 (HNF3alphaM, h, 87X) HMGA2, Hrad17, Ki-67, KRT5, KRT6A, KRT6B, KRT14, Mesothelin / ERC, MMP-7, MMP-11, Mucin5 (MUC5), Nanog, N-myc , NOTCH3, PAX9, PCNA, PDGF-B, PIK3CA, PKCi (Iota), PRDX1, S100A2, S100A4, SOX4, STAT3, TITF1 / TTF1, TP, TS, VEGFR-1, VEGFR-2, WNT1, TRF1, TRF2 A method for genetic testing of lung cancer, wherein the gene is at least one cancer-related gene selected from the group consisting of hTR (TERC) and hTEP1.
16. The cancer-related genes are FGFR4 (Fibroblast growth factor-receptor 4), SERENBP1 (Selenium binding protein1), POSTN (Periostinin, OsteblastPlC). heat shock protein 47), CBP1 (collagen binding protein 1), KDELR3 DEAD (Asp-Glu-Ala-Asp) box polypeptide 17, PTMS (Parathymosine), HIST2H2E Histone 2, H2be), TUSC3 (Tumor suppressor candidate 3), ZNF516 (Zinc fingerer protein in 516), BMI1 Polycombgroupuper ring4, ACAD11, INPP4B 5), CLIP4 (CAP-GLY domain concatenating linker protein family, member 4), AYTL1 Hypothetical protein, RALGPS1 (Ral GEF with PH H3 binding motif 1), PSMB10 [Proteasome (prosome, macropain) subunit, beta1 -FinequipantBench, and LRBA (LPS-responsibleBehitchingBeach 1). -Bisphatase 1), COL6A3 (Collagen, type VI), alpha 3, TM9SF2, Transmembrane 9, superfamily member 2, TAP1, Transporter 1, ATP-bindingca sette, sub-family B (MDR / TAP), DIP2B KIAA1463 protein, TncRNA Trophoblast-derived noncoding RNA, EIF4G1 Eukaryotic translation initiation factor 4 gamma, 1, PRIM2 primase, polypeptide 2A, 58kDa, POR P450 (cytochrome) oxidoreductase, FANCG Fanconi anemia, complementation group G, EXOSC7 Exosome component 7, FDPS Farnesyl diphosphate synthase (farnes l pyrophosphate synthetase, dimethylallyltranstransferase, geranyltranstransferase), C5orf4 Chromosome 5 open reading frame 4, CDCA4 Cell division cycle associated 4, H2AFV H2A histone family, member V, FABP3 Fatty acid binding protein 3, muscle and heart (mammary-derived growth inhibitor), CKLF Chemokine-like factor, MGC4308 Hypothetical pro tein MGC4308, FADS1 Fatty acid desaturase 1, SFTPB Surfactant, pulmonary-associated protein B, PLA2R1 Phospholipase A2 receptor 1,180kDa, SELENBP1 Selenium binding protein 1, FARSB Phenylalanine-tRNA synthetase-like, beta subunit, MEF2D MADS box transcription enhancer factor 2 , Polypeptide D (mycyte enhancer factor 2D), NUTF2 Nuclear transpo t factor 2, ID3 Inhibitor of DNA binding 3, dominant negative helix-loop-helix protein, ID2 Inhibitor of DNA binding 2, dominant negative helix-loop-helix protein, PRG2 Proteoglycan 2, bone marrow, H2AFX H2A histone family, member X , WSB1 WD repeat and SOCS box-containing 1, TRIM33 Tripartite motif-containing 33, RAB5C RAB5C, member RAS oncog ne family, PRIM2 primase, polypeptide 2A, 58kDa, RAB21 RAB21, member RAS oncogene family, SSTK Serine / threonine protein kinase SSTK, ST6GAL1 ST6 beta-galactosamide alpha-2,6-sialyltranferase 1, IRF4 Interferon regulatory factor 4, LRBA LPS- responseive trafficking, beach and anchor anchoring, IL13RA1, Interleukin 13, receptor, alpha 1, ESR1 Estrogen receptor 1, MFAP2 Microfibrillar-associated protein 2, STAT1 Signal transducer and activator of transcription 1,91kDa, PPFIA1 Protein tyrosine phosphatase, receptor type, f polypeptide (PTPRF), interacting protein (liprin), alpha 1, MYO1C Myosin IC, PPFIA4 Protein tyrosine phosphate, receptor type, f polypeptide (PTPRF), interacting From rotein (liprin), alpha 4, RAB31, RAB31, member RAS oncogene family, INPP4B Inositol polyphosphate-4-phosphate, type II, 105 kDa, ESR1 EstrogenMest 6. The method for genetic testing of breast cancer according to claim 5, wherein the gene is at least one cancer-related gene selected.
17. The cancer-related genes are TFF1, COX-2, β-Catenin, c-myc, c-jun, APOC1, YF13H12, CDH17, FUS, APOE, S100A11, GRO1, v-jun, v-raf-1, nibrin, humanin, bcl-2, CAS, semaphorin V, CDK4, CKS1, CKS2, cyclin C, cyclin D, cyclin E, CDC25B, Ki-67, MIA, PCNA, DNA topoisomerase -2a, NE-DIbine, in-DIbing 4, IRF7, HOXB7, NFIL3, SRY-box4and 9, IGF-2, TGF-b3, PLAB, FGF-2, HGF, HDGF, nm23, -ERBB2, C-ERBB3, FGFR-4, IGFR-2, EphB2, H1Histamin receptor, Hemopoetic cell kinase-1, SKY, GRB-2 and 7, p38 MAPK, Axin-2, Transport16; , 25, Transferrin receptor, Cytokeratin-1, catenin-b1and g, matrixmetalloproteases; MMP-1, -2, -3, -7, -10, -11, -12, -14, -16, uPA, catepsin-B , E's-K, LI- and OB-cadherin, E-cadherin, collagenes [I, III, I V, V, VI, VII, XVIII] selected from a3-integrin, fibroblast collagenase inhibitor, S100A4, CD9, IL-8, Osteolectin, Thrombospondin-2, VEGF, and Ostepondinneinternepterinneinternepterinneinternep 6. The method for genetic testing for gastric cancer according to claim 5, wherein the gene is at least one cancer-related gene.
18. The cancer-related genes are CCSA (Colon cancer-specific antigen) -2, CCSA (Colon cancer-specific antigen) -3, CCSA (Colon cancer-specific antigen) -4, CCSA (Colon cancer-specific 5-). CP (Cancer-Placenta) -1, Alpha-catenin, REG1A (Regenerating islet-delivered 1 alpha), DPEP1 (Dipeptidase 1), PAP (Pancreatitis-associated loc) r 1 33.1, FOXQ1 Forkhead box Q1 24.4, MSX2 Msh homeobox homologue 2 22.2, ASCL2 Achaete-scute complex-like 2 17.3, MSX1 Msh homeobox homologue 1 8.5, IRX3 Iroquois homeobox protein3 8. 4, GRHL3 Grainyhead-like 3 7.9, TRIM29 Tripartite motif-continging 29 7.4, ETV4 Ets variant genent 4R (E1A enhancer binding1R4E1A4) bon receptor nuclear translocator-like 2 5.3, TEAD4 TEA domain family member 4 5.2, SP5 Sp5 transcription factor 5.2, HES6 Hairy and enhancer of split64.6, TBX3 T-box 3 4.6, NFE2L3 Nuclear factorREG1B Regenerating islet-delivered 1h 75.8, REG3A Regenerative islet-delivered 3a 29.5, TACSTD2 Tumor-associated calcium signal transducer 21.4 8 Interleukin-8 14.7, SERPINB5 Serpin peptidase inhibitor, clade B, member 5 (Maspin) 13.8, REG1A Regenerating islet-derived 1a 8.2, FAIM2 Fas apoptotic inhibitory molecule 2 7.5, DUSP4 Dual specificity phosphatase 4 7.4, REG4 Regenerating islet-delivered family, member 4.6.8, PHLDA1 Peckstrin homology-like domain, family A, member 1, 6.0, LCN2Li. pocalin 2 (oncogene 24p3) 5.7, RTEL1 Regulator of telomere elongation helicase 1 5.6, TGFBI Transforming growth factor, h induced 5.2, IGFBP2 Insulin-like growth factor binding protein 2 4.8, TDGF1 Teratocarcinoma-derived growth factor 1 4.7, TNFRSF6B Tumor necrosis factor receptor superfamily, member 6b, decoy 4.5, DMBT1 Deleted in malignant brain tumors 1 4.2, TNFRSF10C Tumor necrosis factor receptor superfamily, member 10c, decoy 4.1, ANGPTL1 Angiopoietin-likeDSG4 Desmoglein 4 5.9, CLDN8 Claudin 8 25.8, CDH19 Cadherin 19, type 2 8.3, CEACAM7 Carcinoembryonic antigen-related cell adhesion molecular 8.3, CLDN23 Claudin 23 8.0, NRXN1 Neuroxin 7.1, PCDH19 Protocadherin 19 6.8N GN4X Neuroligin 4, X-linked 6.0, TNXB Tenascin XB 5.6, MUCDHL Mucin and cadherin-like 5.1, PCDH9 Protocadherin 9, 4.9, L1 CAMGIRO-1 TGF-beta, NADH dehydrogenase 2 subunit, Filamine, fibrectin, catepsin H, collagen I a2, collagens (III, IV, V, X), lactadherin, HME, CD24 (. nectadrin), TIMP-1, a6-integrin, MMP-1, -2-3, -7, -11, -13-10, IL-8, Osteolectin, Thrombopondin-2, VEGF, and Osteopontin The genetic test method for colorectal cancer according to claim 5, wherein the gene is at least one cancer-related gene selected from the above.
19. The cancer-associated gene, Fibronectinc X02761; K00799; K02273, Tubulin alpha 1 subunit K00558, Matrix metalloproteinase 14 D26512; X83535, Osteonectin; SPARC J03040, DNA damage repair protein UV excision repair protein; RAD23A D21235, Ubiquitin-conjugating enzyme E2 M74524, Trafficking protein Neutrophil gelatinase-associated lipocalin precursor lipocalin 2 X99133, TRAM protein X63679, ADP / ATP carrier protein J02683, Transcription factor High mobility group protein M23619, Growth factor Insulin-stimulated protein kinase 1 U08316, GTP binding protein Transforming protein rhoA H12 L25080, IFN response Interferon gamma antagonist A25270, Cytokines Macrophage inhibitory cytokine 1 AF019770, Down- egulated genes in HCCs tumor tissues, Immune system IgG, IgG Kb M63438 + U72063, IgG3; IgG1 L; IgG1 K; IgG1 Fcb D78345 + Y14737, IgA1; IGHAb J00220 + S71043, IgC mu heavy chain constant regionb X57086; X57331, IgG receptor FC large subunit P51 U12255, Lymphocyte antigen M81141, Metabolic pathway Betaine-homocysteine S-methyltransferase U50929, Methy enetetrahydrofolate dehydrogenase J04031, Aldehyde oxidase L11005, Uridine diphosphoglucose pyrophosphorylase U27460, Metalloproteinase inhibitor 1 X03124, Metallothionein-IIIb D13365; M93311, Growth factor alpha-2-macroglobulinb M11313, alpha-2-macroglobulin receptor-associated protein M63959, TGF-betac S81439, hepatocyte growth factor-lik protein D49742; S83182, NGF-inducible anti-proliferative protein PC3 U72649, Early growth response protein 1b X52541; M62829, Insulin-like growth factor-binding protein 3c M31159; M35878, Insulin-like growth factor binding protein 4 M62403, IFN response Interferon -Induced protein MXA M33882, Interferon-inducible protein 9-27 J04164, Hormon s, receptors Nuclear hormone receptor L76571, Extracellular matrix Plasminogenb, c X05199, Haemoglobin alpha subunitb, c V00491, Decorin M14219, Tumour associated c-myc oncogene V00568, Cell cycle cyclin-dependent kinase inhibitor 1 U09579; L25610, DNA-binding and chromatin protein DNA-binding protein CPBP U44975 and GTP binding protein Trans Genetic testing method of liver cancer according to claim 5, wherein the at least one cancer-associated genes selected from orming protein rhoB X06820 either group consisting.
20. The cancer-associated gene, CDC28 protein kinase 2 X54942, CDC27HS protein U00001, Extracellular matrix Integrin beta 4 X53587; X52186, Desmoplakin I & II M77830; J05211, Metabolic pathway Procallagen C proteinase M22488 + U50330, Growth factor Platelet-derived growth factor A subunit X06374 , CMP-N-acetylneuraminate-beta-galactosamide-alpha-2,6-sialytransfer se Betaine-homocysteine S-methyl-transferase U50929, Dihydro-orotate dehydrogenaseb M94065, Cytidine deaminase L27943, Aldehyde oxidaseb L11005, Aminoacylase 1 L07548, Methylenetetrahydrofolate dehydrogenaseb J04031, Metallothionein-IIIb X62822, Immune system IgG receptor FC large subunit P51b U12255, Major histocompatibiity complex enhance er-binding protein MAD3 M69043, Leukocyte IgG receptor J04162, FC-epsilon-receptor gamma subunit M33195, HLA-DR antigen-associated invariant subunit X00497, Growth factor Hepatocyte growth factor activator D14012, Growth factor receptor-bound protein 2 isoform L29511; M96995 , Epidical growth factor receptor, oncogene ERBBc X00588; K03193, E F response factor 1 X79067, IFN response STAT-induced STAT inhibitor 2b AB004903, STAT-induced STAT inhibitor 3b, c, Hormone Insulin-induced protein 1b U96876, Cytokine receptor Interleukin-1 receptor antagonist protein precursor M63099, Apoptosis PIG7; gene induced by p53 AF0101212, Growtharrest and DNA-damage-inductive protein M60974, Ce l cycle Cyclin-dependent kinase inhibitor 1; P21 U09579; L25610, G protein Guanine nucleotide-binding protein G U31383, Transcription factor Signal transducer and activator of transcription 3b L29277, Polyhomeotic 2 homolog U89278, Liver specific Haemoglobin alpha subunitb, c V00491, Regulatory protein Gravin M96322, Binding protein DNAX ac tivation protein 12 AF019562, Unknown function KIAA0022 Gene D14664, Growth factor Transforming growth factor, beta-inducedb, Metabolic pathway Lactate dehydrogenase A X02152, Aldehyde oxidaseb L11005, Nucleoside-diphosphate kinase Y07604, Cytoskeleton Alpha 1 cateninb, c D13866; D14705, Beta cateninc X87838; Z19054, Collagen 6 alpha 1 subunit X15879, Type II cytoskeletal 8 keratin M34225, Growth factor, receptor Growth factor receptor-bound protein 2 and 3 isoforms L29511; M96995, Hormone Progesteron receptor-associated protein (HSC70) U28918, Tumour associated c-jun N-terminal kinase 2 L31951, shb proto-oncogene X75342, Transport protein ADP / ATP carrier protein J02683, Immune sy stem FC-epsilon-receptor gamma subunitb M33195, Cytokine receptor Interleukin-1 receptor antagonist protein precursor M63099, Regulatory protein Protein kinase C inhibitor 1 U51004, GTP binding protein Rho GTPase activating protein 4b X78817, Guanine nucleotide-binding protein G (l) alpha subunit M17219, and Genes specially modulated in. At least one HBV according to claim 5, characterized in that the cancer-related genes, or genetic testing method of liver cancer associated with HCV selected from one of the group consisting of CC developed on cirrhotic tissue.
21. The presence risk of the cancer cell population based on a significant difference between the expression level of the cancer-related gene obtained from a blood sample of a healthy person and the expression level of the cancer-related gene obtained from a blood sample of a patient, and / or The method for genetic testing for cancer according to any one of claims 1 to 20, wherein a therapeutic effect is predicted.
22. 21. The risk of cancer cell population and / or therapeutic effect is predicted based on an analysis of the expression level of the cancer-related gene, in which normal expression is not generally observed in healthy persons. A method for genetic testing for cancer according to claim 1.

Images