JP2005304497A - Method for detecting cancer using specified cancer-related gene and method for inhibiting the cancer - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method for detecting cancer comprising finding out a cancer-related gene capable of being used as an indicator for detecting canceration of a cell and malignancy of the cancer and using the cancer-related gene as the indicator, and to provide a method for inhibiting or treating the cancer, using the cancer-related gene as an essential part. <P>SOLUTION: This method for detecting the cancer comprises comprehensively finding out specified genes which are amplified or deleted in various kinds of cancer, such as lung cancer, gastric cancer, pancreatic cancer, colorectal cancer, bile duct cancer, brain tumor, and myeloma, by comparing them with a normal cell, and using amplification or deletion of the cancer-related gene as the indicator. Further, it is found that the cancer is controlled by introducing the gene which is deleted in a cell of the cancer among the cancer-related genes into the cancer and inhibiting a transcriprtional product of the gene which is amplified. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to a detection method capable of detecting canceration and malignancy of cancer using a specific cancer-related gene as an index, and a cancer suppression / treatment method using a specific cancer-related gene as an essential part.

  The death rate from cancer is currently No. 1 in Japan, accounting for about one third of all causes of death. To date, cancer mortality has been increasing and is expected to account for about 50% in 10 years. It has become clear that cancer is caused by accumulation of many gene abnormalities and becomes malignant. It has been reported that increased expression of oncogenes and decreased expression associated with deletion of tumor suppressor genes are involved in carcinogenesis. Furthermore, it is also known that genes directly involved in cell differentiation and proliferation and abnormalities in genes involved in DNA repair systems are involved in canceration.

  However, it is not enough to explain the mechanism of canceration in cancer patients from the results of previous research. The gene group involved in canceration differs depending on the type of cancer, and even if it is the same type of cancer, the individuality of the cancer is different, so the systematic analysis of what gene group abnormality has caused cancer so far It was difficult. Therefore, it is difficult to say that the early diagnosis of cancer by analyzing the genome of cancer cells and the diagnostic means for examining the malignancy of cancer are still sufficiently provided.

  The problem to be solved by the present invention is to find a cancer-related gene that can be used as an index for detecting canceration of cells and malignancy of cancer, and to provide a method for detecting cancer using the cancer-related gene as an index. It is in. Furthermore, an object of the present invention is to provide a method for suppressing or treating cancer using the cancer-related gene as an essential part.

  Normally, when a chromosomal abnormality occurs, the cell is killed by apoptosis, and the abnormal cell does not proliferate. However, for some reason, cells with chromosomal abnormalities pass through the control mechanism of the living body, which is strictly controlled, and cell growth begins, which is the initiation of canceration. Therefore, genome amplification and deletion at the chromosomal level are important factors for canceration. In the case of amplification, the gene expression existing in the amplified genomic region is enhanced, and in the case of deletion, the gene is deleted. It is considered that the expression level of genes existing in the genomic region is markedly reduced, and the accumulation of these abnormalities causes the disordered proliferation of cells.

  Comparative Genomic Hybridization (CGH) is the simplest, fastest and best way to analyze gene abnormalities associated with multiple gene amplifications and deletions on the genome. In order to analyze genetic abnormalities on the genome that are involved in canceration and malignant transformation of cancer, selection of genes to be printed on the CGH microarray is extremely important.

The present inventors selected genes that are highly likely to be involved in canceration from the databases of “National Cancer for Biotechnology” and “University of California Santa Cruz Biotechnology”, and further subjected the selected DNA to a BLAST search to detect cancer. We selected genes that are considered important for the onset. After carefully selecting BAC / PAC clones containing these cancer-related genes as candidates, each was amplified (inexhaustible), and a “MCG cancer array” platform on which more than 800 of these amplified clones were loaded was created. (Hereinafter also referred to as “MCG cancer array”). The present invention includes this MCG cancer array as a technical scope.

  Thus, the present inventors have found one cancer-related gene serving as an index for detecting these cancers in several types of cancers using the MCG cancer array, and completed one of the present inventions.

  That is, the present invention is an invention that provides a cancer detection method (hereinafter also referred to as the present detection method) using a specific cancer-related gene as an index. Cancer-related genes used in this detection method are the following genes.

1. A cancer-related gene in which an amplified copy number is recognized rather than a copy number in the genome of a normal cell (hereinafter also referred to as an amplified oncogene):
MUC1 gene, PRCC gene, EIF4G gene, THPO gene, TERT gene, DAB2 gene, EGFR gene, ELN gene, MUC3 gene, MYC gene, PVT1 gene, FCA gene, PTPN1 gene, Livin gene, MYCN gene, ELK3 gene, BCL2L2 gene , HNF3A gene, ERBB2 gene, CGI-147 gene, ESP15 gene, ARHGEF2 gene, ABCC5 gene, EIF4G1 gene, CDH12 gene, PC4 gene, SKP2 gene, ZNF131 gene, RAD52 gene, WNT3 gene, CDC27 gene, COL1A1 gene, ABCC3 gene , PPM1D gene, ITGB4 gene, BIRC5 gene, SHGC-103396 gene, BCL2L1 gene, RBL1 gene, SRC gene, TGIF2 gene, MYBL2 gene, BIRC7 gene, ARAF1 gene, CUL4B gene, CTAG1 gene, TRIM33 gene, MYCL1 gene, RLF gene , CCNE1 gene, PCTK1 gene, ETV5 gene, CDC10 gene, IGFBP1 gene, TCRG gene, MYCLK1 gene, TAX1BP1 gene, IL6 gene, PMS2 gene, MET residue Gene, SMOH gene, BRAF gene, CDK5 gene, AR gene, MCF2 gene, MAGEA2 gene, CTAG gene, ALX gene, PMF1 gene, NTRK1 gene, ERV5 gene, MUC4 gene, PDGFRA gene, IGFBP7 gene, CDH10 gene, E2F3 gene , TPMT gene, TFAP2A gene, EEF1E1 gene, RREB1 gene, CDK6 gene, PRIM1 gene, GLI gene, CDK4 gene, FUS gene, CYLD gene, GRB2 gene, TGFβR3 gene, PAX3 gene, MLL gene, FKHR gene, FOLR1 gene, PLUNC (LUNX) gene, E2F1 gene, TNFRSF5 gene, NCOA3 gene, ELMO2 gene, NCOA3 (AIB1) gene, PRex1 gene, BCAS1 gene, ZNF217 gene, STK6 (BTAK) gene, SDC1 gene, DNMT3A gene, MLH1 gene, CTNNB1 gene, CCK gene, EGR2 gene, KSAM (FGFR2) gene, PKY (HIPK3) gene, LMO2 gene, CD44 gene, KRAS gene, KRAG (SSPN) gene, CYP1A1 gene, IQGAP1 gene, FURIN ( PACE) gene, PPARBP gene, KRAG gene, KRAS2 gene, PTHLH gene, BCLX gene, DEK gene, Livin-2 gene, TFAP2C gene, TNFRSF6B gene, HCK gene, CDC2L1 gene, GLI3 gene, PPP1A gene, SUPT5H gene, AKT2 gene , TRRAP gene, Smurf1 gene, PDAP1 gene, MIA gene SERPINE1 gene, VGF gene, HRAS gene, BCL3 gene, LUNX gene, AIB1 gene, NCOA gene, SSX4 gene, SSX1 gene, MCL1 gene, CCND3 gene, FLT3 gene, DOC2 gene , BAI1 gene, PSCA gene, MLZE gene, RECQL4 gene, BCL1 gene, FGF4 gene, Survivin gene, BCL9 gene, AF1Q gene, DAP3 gene, BRAL1 gene, TRK gene, CRP gene, ATF6 gene, PBX1 gene, ABL2 gene, LAMC2 Gene, TP gene, ABL gene, CAN gene, NCOR1 gene, ZNF287 gene, D17S128 gene, BCL2 gene, FVT1 gene, PI5 gene, p63 gene, NRG1 Genes, YAP1 genes and cIAP1 genes;

2. Cancer-related genes in which homozygous deletion or heterozygous deletion is observed in genes that should be present in the genome of normal cells due to canceration of cells (hereinafter also referred to as deletion oncogenes):
BAIAP1 gene, LZTS1 gene, AAC1 gene, BLK gene, MTAP gene, CDKN2A (p16) gene, GPC5 gene, SNRPN gene, SAMD4 gene, DCC gene, ADRBK2 gene, DBCCR1 gene, RIZ gene, CCK gene, UBE2E1 gene, THRB gene , RARB gene, TGFβR2 gene, MLH1 gene, CTNNB1 gene, VIPR1 gene, ZNF35 gene, TGM4 gene, TDGF1 gene, PTPRG gene, FHIT gene, MITF gene, LOC151987 gene, EIF4E gene, NFκB gene, CCNA gene, PGRMC2 gene, VEGC Gene, MSH3 gene, RASA1 gene, TPT1 gene, RB1 gene, AATF gene, NR2F1 gene, AFP gene, EGF5 gene, ABCG2 gene, BMI1 gene, PCDH15 gene, PGR gene, FGF9 gene, ZNF198 gene, FLT1 gene, FLT3 gene, BRCA2 gene, KLF12 gene, PIBF1 gene, HNF3A gene, MBIP gene, FKHL1 gene, CDKN2A (p16) gene, N33 gene, TEK gene, MLLT3 gene, JAK 2 genes, GASC1 gene, D9S913 gene, SMAD4 gene, MADH2 gene, MADH7 (SMAD7) gene, MALT1 gene, GRP gene, BCL2 gene, FVT1 gene, SERPINB (PI5) gene, CTDP1 gene, SMAD4-2 gene, D8S504 gene, NAT2 gene, TNFRSF10B gene, MAP3K7 gene, DLC1 gene, stSG42796 gene, LPL gene, NRG1 gene, SCCA1 gene, SCCA2 gene, NKX3A gene, SMAD7 gene, MLL1 gene, PI5 gene, Casp3 gene, SSXT gene, VIM gene, stSG27915 gene , RH68621 gene, SHGC-145820 gene, EEF1E1 gene, ESR1 gene, MLLT3 gene, DEC1 gene, CDH23 gene, ETK1 gene, VIP gene, IGHG1 gene, PMP22 gene, MAFG gene, SHGC-145820 gene, FGF2 gene, ST5 gene, CALCA gene, FKHR gene, CXADR gene, TGFβR3 gene, ABCD3 gene, LCP1 gene, DACH gene, GPC5 (2) gene, GPC6 gene, ABCC4 gene, RAP2A gene , FGF14 gene, ERCC5 gene, EFNB2 gene, ING1 gene, TFDP1 gene, GAS6 gene, D13S327 gene, TEP1 gene, MMP14 gene, BCL2L2 gene, SSTR1 gene, PNN (DRS) gene, CDKN3 gene, RBBP1 gene, DAAM1 gene, MNAT1 Gene, HIF1A gene, ESR2 gene, MAX gene, EIF2S1 gene, RAD51L1 gene, HSXIAPAF1 gene, MN1 gene, TSPY gene, EPS15 gene, YAP1 gene, cIAP1 gene, MMP7 gene, MMP1 gene, DYNEIN gene, ETS1 gene, FLI1 gene, IGHG1 gene, TSPY gene, ABCE1 gene, DMBT1 gene and EGF9 gene;
The cancer-related genes that can be selected as an index vary depending on the type of cancer, and the same cancer-related gene is an amplified oncogene in one cancer, but is a deleted oncogene in the other cancer However, it became clear that it was recognized rarely. Details of these cancer-related genes will be described later.

  In addition, the present invention provides a means for suppressing / treating cancer using the above-mentioned cancer-related gene. Specifically, the present invention relates to a cancer suppression / treatment means by introducing a specific deletion oncogene into a cancer cell, and a cancer by inhibiting the action of a transcription product (mRNA) of a specific amplified oncogene. Provide a means of suppressing and treating The means for suppressing and treating these cancers will also be described later.

A. This detection method This detection method can be performed by, for example, the CGH method, the DNA chip method, the quantitative PCR method, and the real-time RT-PCR method. When detecting gene amplification or deletion, the DNA chip method or the CGH method is preferably performed, and the CGH method is particularly preferable. In addition, for example, the expression of a tumor suppressor gene (corresponding to the above-mentioned “deletion gene”) is caused by gene deletion such as increased methylation of CpG island of the gene or suppression of acetylation of the gene-related protein When it is suppressed by a cause other than the above, it is preferable to use a means for detecting a gene transcript such as a real-time RT-PCR method or a DNA chip method that can quantify the transcript of the gene.

  In addition, the sample that is the subject of this detection method is a sample according to the type of cancer to be detected by the sample provider. That is, for example, in the case of detecting gastric cancer of the sample provider, it is a biopsy sample of the stomach, and in the case of detecting leukemia, it is a blood sample.

  As a particularly preferred embodiment in this detection method, the CGH method is performed by using a plurality of BAC (Bacterial Artificial Chromosome) DNA, YAC (Yeast Artificial Chromosome) DNA, or PAC (Phage Artificial Chromosome) DNA having a specific genomic DNA region. An embodiment using a base in which various types of gene amplification products are established for each gene amplification product can be exemplified, and genomic DNA amplification and deletion gene analysis can be performed by the CGH method.

  Since normally obtained BAC DNA and the like are in a small amount for producing a large number of genomic DNA fixing bases and putting them into practical use, it is necessary to obtain the DNA as a gene amplification product (this gene amplification process is also referred to as “infinite storage”). ). In inexhaustible storage, first, BAC DNA or the like is digested with a 4-base recognition enzyme such as RsaI, DpnI, or HaeIII, and then ligated by adding an adapter. The adapter is an oligonucleotide consisting of 10 to 30 bases, preferably 15 to 25 bases. The double strand has a complementary sequence, and after annealing, the 3′-end oligonucleotide that forms the blunt end is linked to the oligonucleotide. It needs to be oxidized. Next, using a primer having the same sequence part as one of the oligonucleotides of the adapter, it can be amplified by PCR (Polymerase Chain Reaction) method and inexhaustible. On the other hand, a 50-70 base aminated oligonucleotide characteristic of each BAC DNA etc. can also be used as a detection probe.

  The desired DNA fixing base is manufactured by fixing the inexhaustible BAC DNA or the like in this manner (genomic DNA, cDNA, and synthetic oligonucleotides other than the above) on the base, preferably a solid base. can do.

  Examples of the solid substrate include glass, plastic, membrane, and three-dimensional array, and a glass substrate such as a slide glass is preferable. More preferably, the solid substrate such as glass is coated with poly-L-lysine, aminosilane, gold / aluminum, or the like, and the surface is fixed with amino group-modified DNA.

  The concentration at which the above-described inexhaustible DNA (genomic DNA, cDNA, and synthetic oligonucleotide of other embodiments) is spotted on the substrate is preferably 10 pg / μl to 5 μg / μl, more preferably 1 ng / μl to 200 ng / μl. The amount to be spotted is preferably 1 nl to 1 μl, more preferably 10 nl to 100 nl. The size and shape of each spot fixed on the substrate is not particularly limited. For example, the size can be 0.002 to 0.5 mm in diameter, and the shape viewed from the top surface can be circular to elliptical. The thickness of the drying spot is not particularly limited, but is 1 to 100 μm. Further, the number of spots is not particularly limited, but is 10 to 50,000 per substrate to be used, more preferably 100 to 5,000. Each DNA is spotted in the range of Singular to Quadruplicate, but preferably spotted on Duplicate or Triplicate.

  Preparation of the dried spot was performed by, for example, using a spotter for inexhaustible BAC DNA or the like (the same applies to genomic DNA, cDNA, and synthetic oligonucleotide in other embodiments) to form a plurality of spots. Thereafter, the spot can be produced by drying. As the spotter, an ink jet printer, a pin array printer, and a bubble jet (registered trademark) printer can be used, but an ink jet printer is preferably used. For example, GENESOT (Nippon Gaishi Co., Ltd., Nagoya), Cartesian Technologies (USA) high-throughput inkjet dispensing system SQ series, etc. can be used.

  The desired DNA fixing base is manufactured by fixing the inexhaustible BAC DNA or the like in this manner (genomic DNA, cDNA, and synthetic oligonucleotides other than the above) on the base, preferably a solid base. can do. Actually hybridized on the MCG cancer array using Cy-5 labeled genomic DNA derived from the same normal Diploid Cell as the Cy-3 labeled genomic DNA derived from normal Diploid Cell The results (displayed as Merge) are shown in FIG. When Cy-3 labeled genomic DNA is used, it is detected as green, when Cy-5 labeled genomic DNA is used, it is detected as red, and when both are mixed, it is detected as yellow.

In the MCG cancer array shown in FIG. 1, 432 types of BAC DNA were printed. These BAC DNAs comprehensively contain a group of genes related to cancer such as oncogenes and tumor suppressor genes. The size of one section was 1.75 mm long and 2.11 mm wide, and 72 DNAs were printed. There are a total of 432 spots in a vertical row, printed on Duplicate. The results of hybridization with Cy3-labeled normal diploid cell genomic DNA are all green in FIG. 1A. The results of hybridization with Cy5-labeled normal diploid cell genomic DNA are all red in FIG. 1B. Cy3-labeled DNA and Cy5-labeled DNA are mixed and the result of hybridization is all yellow on the slide substrate (FIG. 1C) labeled Merge. When the fluorescence intensity of Cy3 is plotted on the horizontal axis and the fluorescence intensity of Cy5 is plotted on the vertical axis, all signals are almost linear and are aggregated to an intensity of 5 × 10 ³ -5 × 10 ⁴ (FIG. 1D).

In addition, DNA derived from normal cells was labeled with Cy-5, DNA derived from cancer cells was labeled with Cy-3, comparative genomic hybridization was performed, data was captured with a GenePix 4000B scanner, and the results of each pixel were analyzed. This is shown in FIG. The vertical axis of the table in FIG. 2 is represented by Log ₂ Ratio, and the horizontal axis is a BAC clone having a genome from the short arm to the long arm of the chromosome. After correcting the intensity of Cy3 of all spots and the intensity of Cy5 of all spots to the same value, the ratio of Cy3 intensity / Cy5 intensity of each spot is obtained, and the value of Log ₂ Ratio is calculated. BAC having CDKN2A (p16) gene shows a value around Log ₂ Ratio = −3, and Ratio = 1/8, clearly showing homozygous deletion. On the other hand, BAC having the ERBB2 gene has a value of Log ₂ Ratio = 3-4, and Ratio = 8-16, indicating that ERBB2 genomic DNA is amplified 8-16 times.

  In order to identify gene groups present in chromosomal regions that are amplified and deleted in cancer cells using MCG cancer arrays, the genomic DNA derived from healthy individuals and the genomic DNA derived from lung cancer cells are labeled with different dyes. To do. For example, it can be labeled by a conventional method (eg, nick translation method using dCTP) using Cy3, Cy5, or the like. Labeling kits that perform nick translation using dCTP are sold by PanVera (Japan distributor: Takara Shuzo Co., Ltd.), Invitrogen (CA, USA), and the like. When hybridizing labeled DNA with DNA printed on a CGH array, it is more preferable to add Cot-1 DNA, formamide, dextran sulfate, SSC (150 mM NaCl / 15 mM sodium citrate), Yeast t-RNA, and SDS (Sodium Dodecyl Sulfate) . Further, it is preferable to add a solution containing the labeled DNA after heat denaturation. As a container to be hybridized, it is more preferable to use a container that can be placed on a platform with a locking function and can be uniformly contacted on the array using a small amount of solution, for example, a hybrid man. The hybridization temperature is preferably 30 to 70 ° C, more preferably 38 to 45 ° C. The hybridization time is preferably 12 to 200 hours, and more preferably 40 to 80 hours. The array can be washed at room temperature using formamide, SSC solution or the like. The washing of this array is an important step for eliminating nonspecific signals as much as possible. After washing at room temperature, washing is performed at 40-60 ° C. using the same washing solution, and further, 50 ° C. in a solution containing SSC-SDS. More preferably, it is washed with, and left in a solution containing phosphate buffer / NP-40, and finally shaken in a solution containing SSC.

(1) Genes present in chromosomes amplified and deleted in lung cancer cells
Using MCG cancer array, a gene with squamous cell carcinoma, adenocarcinoma, or large cell carcinoma with a ratio value of 1.32 or more, usually 2 copies As a result of investigating the genes on the chromosome that is amplified to 3 copies or more, MUC1 gene, PRCC gene, EIF4G gene, THPO gene, TERT gene, DAB2 gene, EGFR gene, ELN gene, MUC3 gene, MYC gene, PVT1 Gene, FCA gene, PTPN1 gene and Livin gene were detected. Furthermore, the MYCN gene, the MYC gene, the PVT1 gene, the ELK3 gene, the BCL2L2 gene, the HNF3A gene, ERBB2 Gene and CGI-147 gene were detected.

Next, as a gene on the heterozygote chromosome whose ratio value is 0.75 or less, that is, the copy number is halved, BIAAP1 gene, LZTS1 gene, AAC1 gene, BLK gene, MTAP gene, CDKN2A
(p16) Gene, GPC5 gene, SNRPN gene, SAMD4 gene, DCC gene and ADRBK2 gene were detected. Furthermore, CDKN2A (p16) gene, MTAP gene, DBCCR1 gene, and RIZ gene were detected as genes on the chromosome in which homozygote deletion was observed with a Ratio value of 0.25 or less. In the case of DBCCR1 gene, expression suppression by homozygous deletion and expression suppression by CpG island methylation were found.

  Diagnosis of squamous cell carcinoma, adenocarcinoma, or large cell carcinoma can be made by monitoring amplification and deletion of genes detected in this way It is.

  A similar method was used to analyze the genes in the chromosomal region amplified and deleted in non-small cell lung cancer cells. As a result of examining the gene group in which the ratio value is 1.32 or more, that is, the gene group in which 2 copies are usually present is amplified to 3 copies or more, ESP15, MUC1, ARHGEF2, PRCC, ABCC5, EIF4G1, CDH12, PC4, SKP2, DAB2, ZNF131, RAD52, WNT3, CDC27, COL1A1, ABCC3, PPM1D, ITGB4, BIRC5, SHGC-103396, BCL2L1, RBL1, SRC, TGIF2, MYBL2, BIRC7, ARAF1, CUL4B, and CTAG1 genes were detected. The TRIM33, MYCL1, RLF, MYC, CCNE1, and PCTK1 genes were detected as genes having a Ratio value of 4 or more, ie, genes that normally existed in 2 copies were amplified to 8 copies or more.

  Next, as a heterozygous gene whose ratio value is 0.75 or less, that is, the copy number is halved, CCK, UBE2E1, THRB, RARB, TGFβR2, MLH1, CTNNB1, VIPR1, ZNF35, TGM4, TDGF1, PTPRG, FHIT , BIAAP1, MITF, LOC151987, EIF4E, NFκB, CCNA, PGRMC2, VEGFC, MSH3, RASA1, TPT1, RB1, AATF, ADRBK2 genes were detected. Furthermore, the NR2F1 gene was found as a gene with a heterozygote deletion with a Ratio value of 0.25 or less.

  Non-small cell lung cancer can be diagnosed by monitoring amplification and deletion of the chromosomal region having the genes detected in this way

(2) Gene groups present in chromosomes amplified and deleted in brain tumors, etc. Using the same method as in (1) above, a chromosomal region amplified and deleted in malignant glioma is identified, and The existing gene group was analyzed. As a gene amplified in malignant glioma with a ratio of 1.32 or more, EIF4G, ETV5, EGFR, CDC10, IGFBP1, TCRG, MYCLK1, TAX1BP1, IL6, PMS2, MUC3, MET, SMOH, BRAF, CDK5, AR, CUL4B, MCF2, MAGEA2, and CTAG genes are included, and further, the ratio is 4 or more, that is, genes whose gene copy number is amplified 4 times or more of normal cells, ALX, MUC1, ARHGEF2, PMF1, NTRK1, ERV5, MUC4, PDGFRA, IGFBP7, PC4, SKP2, DAB2, CDH10, CDH12, TERT, E2F3, TPMT, TFAP2A, EEF1E1, RREB1, EGFR, PMS2, CDK6, PRIM1, GLI, CDK4, FUS, CYLD, GRB2 genes are found It was.

  As a group of genes deleted in malignant glioma cells, the ratio is 0.75 or less, that is, as heterozygous deletion genes group, AFP, EGF5, ABCG2, NFκB, CDKN2A (p16), MTAP, BMI1, PCDH15, PGR FGF9, ZNF198, FLT1, FLT3, BRCA2, RB1, KLF12, PIBF1, HNF3A, MBIP, and FKHL1 genes were found. Furthermore, the MTAP gene and the CDKN2A (p16) gene were found as the genes whose ratio was 0.25 or less, ie, homozygous deletion was detected.

  Next, as a group of genes amplified in neuroblastoma cells, the MYCN gene is amplified 9-97 times to a very high level, the CDK4 gene is amplified 25 times, and the PPM1D gene is amplified 9.8 times The MYCL1, CDH10, and MYC genes were amplified 2.8-4.2 times.

Next, the gene amplified in rhabdomyosarcoma cells was analyzed. As a result, TGFβR3, PAX3, MLL, CDK4, and FKHR were found as amplified genes as compared with normal cells.

  Diagnosis of brain tumors, especially malignant glioma, Neuroblastoma, Rhabdomyosarcoma is possible by examining the amplification and deletion of the chromosomal region having the genes detected in this way and analyzing the genes that are amplified and deleted It is.

(3) Gene groups present in chromosomes amplified and deleted in gastric cancer Using the same method as in (1) above, the chromosomal region amplified and deleted in gastric cancer cells is identified and present in that region. The gene group to be analyzed was analyzed. As a result of examining the gene group amplified with a ratio of 1.32 or more, PVT1, MYC, FOLR1, PLUNC (LUNX), E2F1, TGIF2, TNFRSF5, NCOA3, ELMO2, MYBL2, NCOA3 (AIB1), PTPN1, PRex1, BCAS1, ZNF217, STK6 (BTAK), CUL4B, MCF2, and CTAG genes were found. SDC1, DNMT3A, MLH1, CTNNB1, CCK, ZNF131, CDK6, MET, MYC, PVT1, EGR2, KSAM are genes whose ratio is 4 or more, that is, 4 times or more of the detected genes compared to normal cells. (FGFR2), PKY (HIPK3), LMO2, CD44, KRAS, KRAG (SSPN), CYP1A1, IQGAP1, FURIN (PACE), PPARBP, ERBB2, CCNE1, and MYBL2 genes were found.

  On the other hand, as a result of examining the deletion of the chromosomal region in gastric cancer cells and analyzing the gene group in that region, the ratio decreased to 0.75 or less, that is, as a gene determined to be a heterozygote, BAIAP1, PTPRG, N33, TEK, MTAP, CDKN2A (p16), MLLT3, JAK2, GASC1, D9S913, SMAD4, MADH2, MADH7 (SMAD7), DCC, MALT1, GRP, BCL2, FVT1, SERPINB (PI5), and CTDP1 genes were found.

  Furthermore, MTAP, CDKN2A (p16), TEK, RB1, and SNRPN genes were found as the genes whose ratio was 0.25 or less, ie, homozygous deletion was detected.

  Diagnosis of gastric cancer is possible by examining the amplification and deletion of the chromosomal region having the gene group thus detected and analyzing the amplified and deleted gene group.

(4) Gene groups present in chromosomes amplified and deleted in pancreatic cancer Using the same method as in (1) above, gene groups present in chromosome regions amplified and deleted in pancreatic cancer cells Identified. As a result of investigating genes on chromosomes with a ratio of 1.32 or higher in pancreatic cancer cells, KRAG, PTPN1, KRAS2, PTHLH, BCLX, DEK, IGFBP1, MYC, Livin-2, PVT1, PRex1, BCAS1, TFAP2C, EGFR, TGIF2, TNFRSF5, TNFRSF6B, EIF4G, PMS2, HCK, MYBL2, ELMO2, PCTK1, CDC2L1, CDC10, TCRG, GLI3, PPP1A, ZNF217, and SRC genes were detected. Genes with a ratio of 4 or more, that is, 4 times more amplification than normal cells, were detected as SUPT5H, AKT2, TRRAP, Smurf1, PDAP1, MYC, PVT1, KRAS2, KRAG, and MIA genes. It was.

On the other hand, as a result of analyzing the gene group present in the chromosomal region deleted in pancreatic cancer cells, the ratio decreased to 0.75 or less, that is, as a gene determined to be a heterozygote, MTAP, DCC, CDKN2A (p16), N33, AAC1, SMAD4-2, GRP, TEK, D8S504, NAT2, LZTS1, TNFRSF10B, D9S913, GASC1, FVT1, MAP3K7, DLC1, MALT1, stSG42796, BAIAP1, BLK, LPL, NRG1, MLLT3, MADH2, SCCA1, SCCA2 NKX3A, SMAD7, MLL1, PI5, Casp3, SSXT, BCL2, JAK2, PTPRG, VIM, stSG27915, RH68621, CTDP1, SHGC-145820, EEF1E1, ESR1, and KLF12 genes were detected.
Furthermore, the CDKN2A (p16), MTAP, N33, MLLT3, TEK, DEC1, CDH23, and SMAD4-2 genes were detected as genes whose ratio was 0.25 or less, that is, homozygous deletion was detected.

  Diagnosis of pancreatic cancer is possible by examining amplification and deletion of the chromosomal region having the gene group thus detected and analyzing the amplified and deleted gene group.

(5) Gene groups present in chromosomes amplified and deleted in colorectal cancer Using the same method as in (1) above, gene groups in the chromosomal region amplified and deleted in colorectal cancer cells were identified. . As a result of examining genes on the chromosome that is amplified with a ratio of 1.32 or more in colorectal cancer cells, ELN, SERPINE1, VGF, MUC3, MYC, PVT1, HRAS, BCL3, BCLX, LUNX, E2F1, TGIF2, HCK, AIB1, PTPN1, NCOA, TNFRSF6B, SSX4, SSX1, ARAF1, CUL4B, CTAG, and MAGEA2 genes were detected. MCL1, CCND3, MYC, PVT1, and FLT3 genes were detected as genes in which the ratio was 4 or more, that is, 4 times or more of amplification was detected as compared with normal cell genes.

  On the other hand, as a result of examining genes on chromosomes that are deleted in colorectal cancer cells, Ratio decreased to 0.75 or less, that is, as genes determined to be heterozygotes, ETK1, MITF, PTPRG, FHIT, RARB, VEGFR, MAP3K7, VIP, N33, D8S504, PCDH15, IGHG1, PMP22, MAFG, SSXT, MADH2, DCC, SMAD4-2, GRP, CTDP1, and SHGC-145820 genes were found. A gene having a Ratio value of 0.25 or less, ie, a homozygous deletion, was not detected from a colon cancer cell line.

  By examining the amplification and deletion of the chromosomal region having the gene group thus detected, and analyzing the amplified and deleted gene group, it is possible to diagnose colorectal cancer.

(6) Gene groups present in chromosomes amplified and deleted in cholangiocarcinoma Using the same method as in (1) above, gene groups present in chromosome regions amplified and deleted in colon cancer cells Identified. As a result of examining genes amplified with a ratio of 1.32 or more, ZNF131, DOC2, DAB2, PC4, SKP2, CDH10, CDH12, TERT, CDK5, BAI1, PSCA, MLZE, RECQL4, BCL1, FGF4, ITGB4, Survivin , SRC, PTPN1, PCTK1, and CTAG genes were detected.

  On the other hand, as a result of examining the deletion of genes in cholangiocarcinoma cells, the ratio was reduced to 0.75 or less in cholangiocarcinoma cells, that is, BIAAP1, PTPRG, TDGF1, EIF4E, NFκB, CCNA, FGF2 , NKX3A, N33, LZTS1, LPL, NRG1, DLC1, BLK, AAC1, NAT2, D8S504, MTAP, JAK2, ST5, CALCA, FLT3, FLT1, and FKHR genes were found. The CXADR gene was found as a gene that showed a gene deletion with a Ratio value of 0.25 or less, that is, a homozygote deletion.

  Bile duct cancer can be diagnosed by examining amplification and deletion of the chromosomal region having the gene group thus detected and analyzing the amplified and deleted gene group.

(7) Gene groups present in chromosomes amplified and deleted in myeloma Using the same method as in (1) above, gene groups present in chromosomal regions amplified and deleted in myeloma cells Identified. As a result of examining genes amplified with a ratio of 1.32 or more, BCL9, MCL1, AF1Q, MUC1, DAP3, ARHGEF2, PMF1, BRAL1, PRCC, TRK, NTRK1, CRP, ATF6, PBX1, ABL2, LAMC2, TP A gene was detected. ABL, CAN, KRAS2, KRAG, PTHLH, NCOR1, ZNF287, D17S128, BCL2, FVT1, and PI5 genes are genes that have a ratio of 4 or more, that is, 4 times more amplified than normal cells. was detected.

  On the other hand, as a result of analyzing gene deletion in myeloma cells, the ratio was reduced to 0.75 or less, that is, the genes determined to be heterozygotes were TGFβR3, ABCD3, ZNF198, FGF9, FLT3, FLT1, BRCA2, FKHR, TPT1. , LCP1, RB1, DACH, PIBF1, KLF12, GPC5, GPC5 (2), GPC6, ABCC4, RAP2A, FGF14, ERCC5, EFNB2, ING1, TFDP1, GAS6, D13S327, TEP1, MMP14, BCL2L2, FKHL1, MBIP, HNF3A SSTR1, PNN (DRS), CDKN3, RBBP1, DAAM1, MNAT1, HIF1A, ESR2, MAX, EIF2S1, RAD51L1, IGHG1, HSXIAPAF1, MN1, and TSPY genes were detected. EPS15, PGR, YAP1, cIAP1, MMP7, MMP1, DYNEIN, ETS1, FLI1, IGHG1, and TSPY genes were detected as genes with a ratio of 0.25 or less, that is, homozygous deletion.

Bile duct cancer can be diagnosed by examining amplification and deletion of the chromosomal region having the gene group thus detected and analyzing the amplified and deleted gene group.

(8) Gene groups present in chromosomes amplified and deleted in anaplastic thyroid cancer Amplified and deleted in anaplastic thyroid cells (thyroid cancer cells) using the same method as in (1) above. A group of genes present in the chromosomal region was identified. As a result of investigating genes amplified with a Ratio of 1.32 or more, genes p63, TERT, DOC2, PPP1A, E2F1, and AIB1 were detected. The MET, NRG1, MYC, PVT1, YAP1, and cIAP1 genes were detected as genes in which the ratio was 4 or more, that is, 4 times or more of amplification was detected as compared with the normal cell genes.

  On the other hand, as a result of examining the deletion of the gene in Anaplastic thyroid cells, the ratio decreased to 0.75 or less, that is, as genes determined to be heterozygotes, CTNNB1, BAIAP1, ABCE1, ESR1, N33, LZTS1, DMBT1, EGF9, PMP22 , SMAD4-2 and ADRBK2 genes were detected. Furthermore, MLLT3 and CDKN2A (p16) genes were found as genes with a ratio of 0.25 or less, that is, homozygous deletion was detected.

  Diagnosis of thyroid cancer is possible by examining the amplification and deletion of the chromosomal region having the gene group thus detected and analyzing the amplified and deleted gene group.

  As described above, amplification and deletion of lung cancer, brain tumor, stomach cancer, pancreatic cancer, colon cancer, bile duct cancer, myeloma, etc. by analyzing the amplification and deletion of the chromosomal region using the MCG cancer array Identified genes can be identified. From these results, it is possible to understand the situation of each cancer. That is, first, it is possible to determine whether a tumor is a benign tumor, an intermediate type, or a malignant tumor. In the case of a malignant tumor, it is possible to provide important knowledge in determining the grade of cancer. Furthermore, it is possible to provide data for performing effective treatment in performing postoperative chemotherapy after removing a cancerous part by surgery.

  In addition, as described above, the DBCCR1 gene was found not to cause gene expression due to homozygous deletion, and to the case where the expression was not detected because the CpG island of the gene was not deleted but was deleted. . For these reasons, DBCCR1 gene expression is suppressed with high probability in non-small cell lung cancer.

  As described above, for the deletion oncogene group, gene expression can be monitored by a real-time RT-PCR method, a DNA chip method, etc., and it is possible to examine chromosome deletion and expression suppression at the same time. is there.

B. Cancer Suppression / Treatment Means Using Cancer-Related Genes The cancer suppression / treatment means provided by the present invention includes (1) a gene (deletion oncogene) in which gene deletion is associated with canceration of cells, cancer cells A method for suppressing the cancer cell (hereinafter also referred to as suppression / treatment means 1), and (2) a gene amplification associated with a cell canceration gene (amplified oncogene). Thus, the method can be roughly divided into methods for inhibiting cancer cells by causing them to act on cancer cells (hereinafter also referred to as suppression / treatment means 2).

(1) Inhibition / treatment 1
Among the above-described deletion oncogenes, many genes in the category of tumor suppressor genes were detected in the genes of the chromosomal region showing homozygote deletion. Among them, a gene that suppresses the growth of the target cancer or a gene that induces cancer death by apoptosis can be introduced using a Sendai virus vector or an adenovirus vector. As a promoter of a homozygous deletion gene to be expressed in gene therapy using these viral vectors, a promoter that is highly expressed in cancer tissue and extremely low in normal tissue, for example,
Human CXCR4 promoter (Zhu ZB, Makhija SK, Lu B, Wang M, Kaliberova L, Liu B, Rivera AA, Nettelbeck DM, Mahasreshti PJ, Leath CA, Yamaoto M, Alvarez RD, Curiel DT: Transcriptional targeting of adenoviral vector through the CXCR4 tumor-specific promoter, Gene ther., 11 , 645-648, 2004), Survivin promoter, etc. are preferably used. These recombinant viruses can also be introduced into cancer tissue by forming a complex with liposomes. It can also be introduced into cancer tissue in the form of naked DNA.

  Using the viral vectors and promoters described above, the LRP1B gene localized at 2q22.1 for the treatment of esophageal cancer, the MTAP gene localized at 9p21 for non-small cell lung cancer (Squamous cell carcinoma, Adenocarcinoma, Large cell carcinoma) and CDKN2A (p16) gene, DBCCR1 localized at 9p34, RIZ gene localized at 1p36, NR2F1 gene localized at 5q15 in small cell lung cancer, MTAP gene localized at 9p21 and CDKN2A (p16) in malignant glioma In gastric cancer, MTAP gene localized at 9p21 and CDKN2A (p16) gene, TEK gene localized at 9p21.2, RB1 gene localized at 13q14.2, SNRPN gene localized at 15q11.2, In pancreatic cancer, N13 gene localized at 8p22, MTAP gene localized at 9p21 and CDKN2A (p16) gene, TEK gene localized at 9p21.2, MLLT3 gene localized at 9p22, DEC-localized at 9q32 1 gene, CDH23 gene located at 10q22.1, SMAD4-2 located at 18q21 CXADR gene localized at 21q11.2 in cholangiocarcinoma, EPS15 gene localized at 1p32 in myeloma, PGR localized at 11q22, cIAP1, MMP1, DYNEIN gene, YAP1 localized at 11q22.1 MMP7 gene localized at 11q21-q22, ETS1 gene localized at 11q23.3, FLI1 gene localized at 11q24, IGHG1 gene localized at 14q32.33, TSPY gene localized at Ypter-p11.2 In thyroid cancer, the CDKN2A (p16) gene localized at 9p21 and the MLLT3 gene localized at 9p22 are candidate genes, and can be selected from them.

  The CDKN2A (p16) gene is a cyclin dependent kinase inhibitor located on chromosome 9p21 and is considered a tumor suppressor gene. p16 protein binds to CDK4 kinase to suppress its activity and suppress Cell Cycle progression. The CDKN2A (p16) gene is deleted in a wide range of cancers including acellular esophageal cancer, malignant glioma, gastric cancer, pancreatic cancer, and thyroid cancer. MTAP is a gene encoding 5'-methylthioadenosine phosphorylase and is the first enzyme of methionine salvage pathway and is considered to be a tumor suppressor gene. Products of the methionine salvage pathway inhibit the activity of ornithine decarboxylase, which is highly expressed in cancer. RIZ is a gene encoding the RB interacting Zinc Finger protein found in leukemia and belongs to the Nuclear protein methyltransferase superfamily. DBCCR1 is found as a gene deleted in chromosome 1 of bladder cancer and is considered a tumor suppressor gene. TEK is an Angiopoietin-1 receptor, also known as Tie-2, and angiogenesis is induced by phosphorylation by Tyrosin kinase. CDH23 is a Cadherin related 23 gene, belongs to the Cadherin superfamily, and is a glycoprotein involved in calcium-dependent cell adhesion. The CXADR gene encodes coxsachie virus and adenovirus receptors. The cIAP1 gene encodes an apoptosis inhibitor. The FLI1 gene is classified as an ETS transcription factor. The TSPY gene is present on the human Y chromosome and encodes Testis specific protein. LRP1B is an abbreviation for Lipoprotein receptor-related protein 1B, which is a cell membrane receptor having urokinase, plasminogen activator, etc. as ligands, and is considered a tumor suppressor gene. DEC1 stands for Deleted in esophageal cancer 1, and Loss of heterozygosity is often detected in esophageal cancer and squamous cell carcinoma of the bladder, lung and head and neck. MMP1 and MMP7 are Matrix metalloproteinases, enzymes involved in angiogenesis. The SMAD4 gene is a tumor suppressor gene that has been found to be deleted in pancreatic cancer, and encodes a protein that is activated by a receptor and translocates to the nucleus to induce transcriptional activation. ETS1 is a transcription factor and induces the Angiopoietin-2 gene and the like. RB1 is a Retinoblastoma gene and a tumor suppressor gene.

  A viral vector is prepared by incorporating these genes downstream of a promoter that is highly expressed in cancer tissue, and introduced into the cancer tissue of a cancer patient. By expressing the gene, it is possible to prevent cancer shrinkage and inhibition of cancer metastasis, that is, recurrence after cancer removal.

(2) Inhibition / treatment 2
In addition, among the amplified oncogenes found above, Table 1 shows a group of genes present in chromosomes that are amplified four times or more in cancer cells compared to normal cells.

  Compared with normal cells, the number of copies of these genes increased to 8 or more copies on chromosome 1-22 and 4 or more copies on X and Y chromosomes. These high-expressing transcripts can be treated for cancer by using RNAi (RNA interference) techniques to add small interference RNA to the corresponding transcript (mRNA), leading to degradation of the transcript. Is possible. The siRNA design, synthesis, cell transfection method, and RNAi effect confirmation method can be performed using conventional methods. For example, Takara Bio RNAi BOOK <Experimental protocol collection> (Takara Bio Inc., Shiga Prefecture) Can be referred to. Examples of siRNA that can be used here include Hairpin siRNA that can be expressed using siRNA oligonucleotides and pSilencer vector (Funakoshi Co., Ltd., Tokyo).

On the other hand, it is also possible to knock out the oncogene mRNA that is amplified and overexpressed in cancer cells using antisense oligonucleotides. In this case, s-oligonucleotide has better intracellular stability than ordinary oligonucleotides, and it is more preferable to examine the inhibition of proliferation of cancer cells. SiRNA, Hairpin siRNA, s-ol found to be effective in cancer cells
About igonucleotide, it can evaluate using the nude mouse which transplanted the cancer cell.

  In this case, it is preferable to construct a delivery system so that these RNAs accumulate in cancer tissue.

  According to the present invention, a cancer-related gene that can be used as an index for detecting canceration of cells and the malignancy of cancer is found, and a method for detecting cancer using the cancer-related gene as an index is provided. Cancer suppression and treatment methods using related genes as essential parts have been provided.

[Example 1] Creation of "MCG cancer array"
National Cancer for Biotechnology and University of California Santa Cruz Biotechnology genome database websites and BAC / PAC clones with genes or Sequence Tagged Site markers that are extremely important for canceration and cancer cell growth from the results of BLAST searches of selected DNA Selected.

  BAC and PAC DNA were digested with DpnI, RsaI, and HaeIII, and then ligated with adapter DNA. Next, PCR was performed twice using a primer having an adapter sequence. One of the two primers is aminated at the 5 'end. This process is called inexhaustible storage, and the obtained DNA is defined as inexhaustible DNA. Inexhaustible DNA was covalently printed on an oligo DNA microarray (Matsunami Glass, Osaka) on Duplicate using an inkjet-type spotter (GENESHOT, NGK, Nagoya).

[Example 2] Comprehensive analysis of cancer-related genes in non-small cell lung cancer using MCG cancer array Genes amplified and deleted in 27 types of non-small cell lung cancer cell lines using "MCG cancer array" I investigated. The 27 types are: 11 strains of squamous cell carcinoma, 10 strains of Adenocarcinoma, and 6 strains of large cell carcinoma. Genomic DNA was prepared from these cells. As a control, male healthy human genomic DNA was used. Nick translation of DpnI digested genomic DNA (0.5mg) in the presence of 0.2mM dATP, 0.2mM dTTP, 0.2mM dGTP, 0.1mM dCTP and 0.4mMCy3-dCTP (lung cancer cells) or 0.4mMCy5-dCTP (normal cells), respectively Labeled with Cy3 and Cy5 labeled dCTP were obtained from Amersham Biosciences (Tokyo). Both labeled genomic DNAs are precipitated by adding ethanol in the presence of Cot-1 DNA (Invitrogen), and 120 μl of hybridization mixture (50% formamide, 10% Dextran sulfate, 2X SSC (1xSSC: 150 mM NaCl / 15 mM Sodium)
Citrate), 4% sodium dodecyl sulfate, pH 7). After incubation at 37 ° C. for 30 minutes, the mixture was added to the CGH array set in the hybridization chamber, and incubated at 37 ° C. at a speed of 3 rpm (round per minute) for 48-72 hours. The CGH array was then washed in 50% formamide / 2x SSC (pH 7.0) solution at 50 ° C for 15 minutes, then washed in 2xSSC / 0.1% SDS at 50 ° C for 15 minutes, and then 0.1% The plate was washed with 0.1 M phosphate buffer (pH 8) containing Nonidet P-40 at room temperature for 15 minutes. After air drying, the CGH array was monitored for fluorescence derived from Cy3 and Cy5 using a GenePix 4000B scanner (Axon Instruments, CA, USA) (FIG. 1). The obtained results were analyzed using GenePix Pro4.1 imaging software (Axon Instruments). The average fluorescence intensity derived from Cy3 and the average fluorescence intensity derived from Cy5 were adjusted to the same value, and the ratio of Cy3 / Cy5 was determined. The Ratio value is 1 when there is no abnormality in the genome. It was determined that genomic amplification occurred when the Ratio value was 1.32 or more, and significant amplification was observed when the Ratio value was 4 or more. When the Ratio value was 0.75 or less, it was judged that the possibility of genomic heterozygote deletion was extremely low, and when the ratio value was 0.25 or less, the possibility of homozygote deletion was extremely high.

As shown in Table 2, as a gene amplified in a squamous cell carcinoma (SCC cell), Adenocarcinoma (AdC cell) and Large cell carcinoma (LCC cell) with a Ratio value of 1.32 or more (Log ₂ Ratio value of 0.4 or more) MUC1 gene, PRCC gene, EIF4G gene, THPO gene, TERT gene, DAB2 gene, EGFR gene, ELN gene, MUC3 gene, MYC gene, PVT1 gene, FCA gene, PTPN1 gene and Livin gene were detected.

On the other hand, MYCN gene localized in chromosome 2p24 in 1 cell line, MET gene localized in chromosome 7q23 in 2 cell lines, as a gene amplified to a ratio value of 4 or more (Log ₂ Ratio value of 2 or more), MYC gene localized to chromosome 8q24 in 4 cell lines, PVT1 gene localized to chromosome 8p24 in 4 cell lines, ELK3 gene localized to chromosome 12q23 in 1 cell line, BCL2L2 localized to chromosome 14q11 in 1 cell line Gene, HNF3A gene localized in chromosome 14q12 in one cell line, ERBB2 gene localized in chromosome 17q12 in one cell line, and CGI-147 gene localized in chromosome 17q22 in one cell line were detected.

As shown in Table 3, the BAIAP1 gene is a gene that has a ratio value of 0.75 or less (Log ₂ Ratio value of -0.4 or less) and a copy number of half (heterozygote) in squamous cell carcinoma, Adenocarcinoma, and large cell carcinoma. LZTS1 gene, AAC1 gene, BLK gene, MTAP gene, p16 gene, GPC5 gene, SNRPN gene, SAMD4 gene, DCC gene, ADRBK2 gene were found. Furthermore, the CDKN2A (p16) gene present in chromosome 9p21 in 6 cell lines and the chromosome in 4 cell lines are genes in which the homozygote deletion was observed with a Ratio value of 0.25 or less (Log ₂ Ratio value of -2.0 or less) The MTAP gene present in 9p21, the RIZ gene present in chromosome 1p36 in one cell line, and the DBCCR1 gene present in chromosome 9p34 in one cell line were detected.

[Example 3] Comprehensive analysis of cancer-related genes in small cell lung cancer (SCLC) using MCG cancer array Gene amplification was analyzed using MCG cancer array for small cell lung cancer. As a result, as shown in Table 4, about half of the cell lines tested were ESP15, MUC1, ARHGEF2, PRCC, ABCC5, EIF4G1, CDH12, PC4, SKP2, DAB2, ZNF131, RAD52, WNT3, CDC27, COL1A1 , ABCC3, PPM1D, ITGB4, BIRC5, SHGC-103396, BCL2L1, RBL1, SRC, TGIF2, MYBL2, BIRC7, ARAF1, CUL4B, and CTAG1 genes were amplified at a ratio of 1.32 or higher.

  Further, as shown in Table 5, TRIM33, MYCL1, RLF, MYC, CCNE1 are genes in which the ratio is 4 or more in small cell lung cancer cells, that is, the amplification of 4 times or more compared to the gene of normal cells. The PCTK1 gene was found.

  Next, the ratio is reduced to 0.75 or less in non-small cell lung cancer cells, i.e., CCK, UBE2E1, THRB, RARB, TGFβR2, MLH1, CTNNB1, VIPR1, ZNF35, TGM4, TDGF1, PTPRG , FHIT, BAIAP1, MITF, LOC151987, EIF4E, NFκB, CCNA, PGRMC2, VEGFC, MSH3, RASA1, TPT1, RB1, AATF, and ADRBK2 genes were detected (Table 6).

  Furthermore, the NR2F1 gene was found as a gene in which the ratio was 0.25 or less, that is, a homozygous deletion was detected (Table 7). These heterozygous and homozygous gene groups are likely to cause canceration due to a marked decrease in the expression level.

[Example 4] Comprehensive analysis of cancer-related genes in malignant glioma using MCG cancer array [ MCG Cancer Array ] was used to examine genes amplified and deleted in 22 malignant glioma cell lines. The 22 cell line names used here are KNS-42, KNS-60, KNS-81, KNS-89, No. 10, No. 11, KINGS-1, T98G, GB-1, KS-1, and SF126. , Marcus, Becker, AM-38, A-172, KALS-1, YH-13, YKG-1, U251MG, NMC-G1, U87MG, U373MG. As a result of analyzing the change in the number of genome copies in the malignant glioma cell line, the ratio is a value of 1.32 or more, and as a gene for which amplification was detected, EIF4G, ETV5, EGFR, CDC10, IGFBP1, TCRG, MYCLK1, TAX1BP1, IL6, PMS2, MUC3, MET, SMOH, BRAF, CDK5, AR, CUL4B, MCF2, MAGEA2, and CTAG genes were found (Table 8). Amplification of these genes was detected in about half of the glioma cell lines tested.

  Furthermore, the genes whose ratio is 4 or more, that is, the gene copy number is amplified 4 times that of normal cells, are ALX, MUC1, ARHGEF2, PMF1, NTRK1, ERV5, MUC4, PDGFRA, IGFBP7, PC4, SKP2, DAB2 , CDH10, CDH12, TERT, E2F3, TPMT, TFAP2A, EEF1E1, RREB1, EGFR, PMS2, CDK6, PRIM1, GLI, CDK4, FUS, CYLD, GRB2 genes were found (Table 9). The number of cells in which high level amplification was found is 1-2.

  As a result of analyzing the increase in the number of copies in the chromosome and the expression level of the SKP2 gene, they were correlated (FIG. 3).

  In FIG. 3, normal cells (Normal) and small cell lung cancer cells (S-2, SBC-5, ACC-LC-5, ACC-LC-172, ACC-LC-80, ACC-LC-173, Lu -143, Lu-24, Lu-130, Lu-134A, Lu-138), the SKP2, PC4 and CDH6 genes were amplified by the Southern blot method, and the expression levels of SKP2 mRNA and PC4 mRNA were analyzed by the Northern blot method. Results are shown. GAPDH was used for Northern blot control. In addition, spots where gene amplification and increased expression level were observed are indicated by *.

  Furthermore, the addition of SKP2 antisense oligonucleotide to the PC-10 cell culture medium inhibited Invasion and Migration of PC-10 cells. This confirms that increased SKP2 gene expression is likely to be involved in the acquisition of malignant glioma traits.

  On the other hand, the ratio of malignant glioma cells with a ratio of 0.75 or less, that is, heterozygous deletion was detected as AFP, EGF5, ABCG2, NFκB, p16, MTAP, BMI1, PCDH15, PGR, FGF9, ZNF198, FLT1, FLT3, BRCA2, RB1, KLF12, PIBF1, HNF3A, MBIP, and FKHL1 genes were found (Table 10). Heterozygous deletions of these genes were found in about half of the glioma cell lines tested.

  Furthermore, the MTAP gene and the CDKN2A (p16) gene were found as the genes whose ratio was 0.25 or less, ie, homozygous deletion was detected (Table 11). These heterozygous and homozygous gene groups may cause canceration due to a marked decrease in the expression level.

  In particular, homozygous deletion genes are tumor suppressor genes, and deletion of these gene groups plays an important role in inducing canceration.

[Example 5] Analysis of amplified genes in neuroblastoma (Neuroblastoma) and rhabdomyosarcoma ( MCG cancer array ) Using the "MCG cancer array", the genes that were amplified were analyzed. . As a result, in comparison with normal cells, the MYCN gene localized on chromosome 2p24 was detected at 9-97-fold in 15 cell lines, and a very high level of amplification was detected (Table 12). Furthermore, the CDK4 gene localized on chromosome 12q14 was amplified 25 times and the PPM1D gene localized on chromosome 17q23 was amplified 9.8 times. It is very likely that significant amplification of these genes is closely related to canceration and progression of cancer malignancy. MYCL1, CDH10, and MYC genes were amplified 2.8-4.2 times.

  Using the "MCG cancer array", the amplified genes were analyzed for 7 Rhabdomyosarcoma (4 Embryonal cell lines and 3 Alveolar cell lines). As a result, compared to normal cells, TGFβR3, PAX3, MLL, CDK4, and FKHR were detected as amplified genes (Table 13). In particular, the CDK4 gene is amplified 15-fold in the Rh30 cell line and is considered to play an important role in this cell line.

[Example 6] Comprehensive analysis of cancer-related genes in gastric cancer using MCG cancer array Using the "MCG cancer array", the gene amplified or deleted was analyzed for 31 gastric cancer cell lines. The breakdown of 31 shares is 9 well-differentiated adenocarcinoma, 19 undifferentiated adenocarcinoma (8 Poorly differentiated adenocarcinoma and 8 signet)
ring cell carcinoma (including 11 strains), Adenosquamous carcinoma (1 strain), and 2 unidentified strains. Table 14 shows the names, histology, and cell origin of 31 strains of gastric cancer cells.

  As a result of investigating the genes amplified in a ratio of 1.32 or more for 31 gastric cancer cell lines, PVT1, MYC, FOLR1, PLUNC (LUNX), E2F1, TGIF2, TNFRSF5, NCOA3, ELMO2, MYBL2, NCOA3 ( AIB1), PTPN1, PRex1, BCAS1, ZNF217, STK6 (BTAK), CUL4B, MCF2, and CTAG genes were found (Table 15). Amplification of these genes was detected in 58-75% of the gastric cancer cell lines tested.

  Furthermore, as shown in Table 16, SDC1, DNMT3A, MLH1, CTNNB1, CCK, ZNF131 are genes in which the ratio is 4 or more in gastric cancer cells, that is, 4 or more times higher than that of normal cells. , CDK6, MET, MYC, PVT1, EGR2, KSAM (FGFR2), PKY (HIPK3), LMO2, CD44, KRAS, KRAG (SSPN), CYP1A1, IQGAP1, FURIN (PACE), PPARBP, ERBB2, CCNE1, MYBL2 genes It was found. Amplification of these gene groups tended to be higher in differentiated cell lines than in highly differentiated cell lines.

Next, BIAAP1, PTPRG, N33, TEK, MTAP, CDKN2A (p16), MLLT3, JAK2, GASC1, D9S913, SMAD4, MADH2 are genes whose ratio is reduced to 0.75 or less in gastric cancer cells, that is, heterozygotes. MADH7 (SMAD7), DCC, MALT1, GRP, BCL2, FVT1, SERPINB (PI5), and CTDP1 genes were found (Table 17).

  Furthermore, MTAP, CDKN2A (p16), TEK, RB1, and SNRPN genes were found as genes whose ratio was 0.25 or less, ie, homozygous deletion was detected (Table 18). These heterozygous and homozygous gene groups may cause canceration due to a marked decrease in the expression level.

  In particular, homozygous deletion genes are tumor suppressor genes, and deletion of these gene groups plays an important role in inducing canceration.

[Example 7] Comprehensive analysis of cancer-related genes in pancreatic cancer using MCG cancer array Using "MCG cancer array", genes amplified or deleted were analyzed for pancreatic cancer cells. As a result of investigating genes amplified with a ratio of 1.32 or more, KRAG, PTPN1, KRAS2, PTHLH, BCLX, DEK, IGFBP1, MYC, Livin-2, PVT1, PRex1, BCAS1, TFAP2C, EGFR, TGIF2, TNFRSF5 , TNFRSF6B, EIF4G, PMS2, HCK, MYBL2, ELMO2, PCTK1, CDC2L1, CDC10, TCRG, GLI3, PPP1A, ZNF217, and SRC genes were found (Table 19). Amplification of these genes was detected in 50-64% of the pancreatic cancer cell lines tested.

Pancreatic cancer cells with a ratio of 4 or more, that is, 4 times more amplified than normal cells, SUPT5H, AKT2, TRRAP, Smurf1, PDAP1, MYC, PVT1, KRAS2, KR
AG and MIA genes were found (Table 20). High level amplification of these genes was found in 4-16% of cell lines.

  Next, the ratio of the pancreatic cancer cells decreased to 0.75 or less, i.e., genes determined as heterozygotes, MTAP, DCC, CDKN2A (p16), N33, AAC1, SMAD4-2, GRP, TEK, D8S504, NAT2, LZTS1, TNFRSF10B, D9S913, GASC1, FVT1, MAP3K7, DLC1, MALT1, stSG42796, BAIAP1, BLK, LPL, NRG1, MLLT3, MADH2, SCCA1, SCCA2, NKX3A, SMAD7, MLL1, PI5, Casp2, SSXTB, CLasp2, SSXTB PTPRG, VIM, stSG27915, RH68621, CTDP1, SHGC-145820, EEF1E1, ESR1, and KLF12 genes were found (Table 21). Heterozygous deletion of these genes was frequently detected in 50-82% of pancreatic cancer cell lines tested.

  Further, CDKN2A (p16), MTAP, N33, MLLT3, TEK, DEC1, CDH23, SMAD4-2 genes were found as the genes whose ratio was 0.25 or less, that is, homozygous deletion was detected (Table 22). . These heterozygous and homozygous gene groups may cause canceration due to a marked decrease in the expression level.

[Example 8] Comprehensive analysis of cancer-related genes in colorectal cancer using MCG cancer array Using "MCG cancer array", genes amplified and deleted were analyzed for colorectal cancer cell lines. As a result of examining genes amplified with a ratio of 1.32 or more, ELN, SERPINE1, VGF, MUC3, MYC, PVT1, HRAS, BCL3, BCLX, LUNX, E2F1, TGIF2, HCK, AIB1, PTPN1, NCOA, TNFRSF6B , SSX4, SSX1, ARAF1, CUL4B, CTAG, and MAGEA2 genes were found (Table 23). Amplification of these genes was detected in 54-68% of the colon cancer cell lines tested.

  MCL1, CCND3, MYC, PVT1, and FLT3 genes were found as genes in which the ratio was 4 or more in colon cancer cells, that is, 4 times or more of amplification was detected compared to normal cells (Table 24). . High level amplification of these genes was found in 9-18% cell lines.

  Next, the ratio of the colon cancer cells decreased to 0.75 or less, that is, the genes determined to be heterozygotes, such as ETK1, MITF, PTPRG, FHIT, RARB, VEGC, MAP3K7, VIP, N33, D8S504, PCDH15, IGHG1, PMP22 , MAFG, SSXT, MADH2, DCC, SMAD4-2, GRP, CTDP1, and SHGC-145820 genes were found (Table 25). Heterozygous deletion of these genes was frequently detected in 54-82% of the colon cancer cell lines tested.

  A gene having a Ratio value of 0.25 or less, ie, a homozygous deletion, was not detected from a colon cancer cell line.

[Example 9] Comprehensive analysis of cancer-related genes in cholangiocarcinoma using MCG cancer array Using the "MCG cancer array", genes amplified and deleted were analyzed for cholangiocarcinoma cell lines. As a result of examining genes amplified with a ratio of 1.32 or more, ZNF131, DOC2, DAB2, PC4, SKP2, CDH10, CDH12, TERT, CDK5, BAI1, PSCA, MLZE, RECQL4, BCL1, FGF4, ITGB4, Survivin , SRC, PTPN1, PCTK1, and CTAG genes were found (Table 26). Amplification of these genes was detected in 50-75% of the 8 cholangiocarcinoma cell lines tested.

  Next, the ratio of cholangiocarcinoma cells decreased to 0.75 or less, i.e., genes determined to be heterozygotes, BIAAP1, PTPRG, TDGF1, EIF4E, NFκB, CCNA, FGF2, NKX3A, N33, LZTS1, LPL, NRG1, DLC1 BLK, AAC1, NAT2, D8S504, MTAP, JAK2, ST5, CALCA, FLT3, FLT1, and FKHR genes were found (Table 27). Heterozygous deletion of these genes was frequently detected in 4-7 strains of 8 cholangiocarcinoma cell lines tested.

  A CXADR gene localized in 21q11.2 was found in one cell line as a gene showing a gene deletion with a Ratio value of 0.25 or less, ie, a homozygote deletion.

[Example 10] Comprehensive analysis of cancer-related genes in Myeloma (myeloma) using the MCG cancer array Using the "MCG cancer array", the amplified and deleted genes were analyzed for the Myeloma cell line . As a result of examining genes amplified with a ratio of 1.32 or more, BCL9, MCL1, AF1Q, MUC1, DAP3, ARHGEF2, PMF1, BRAL1, PRCC, TRK, NTRK1, CRP, ATF6, PBX1, ABL2, LAMC2, TP Genes were found (Table 28). Amplification of these genes was detected as frequently as 63-82% of the Myeloma cell lines tested.

  Genes with a ratio of 4 or more in Myeloma cells, that is, 4 times or more compared to normal cells, were detected as ABL, CAN, KRAS2, KRAG, PTHLH, NCOR1, ZNF287, D17S128, BCL2, FVT1, The PI5 gene was found (Table 29).

  Next, the ratio of Myeloma cells decreased to 0.75 or less, that is, as a gene determined to be a heterozygote, TGFβR3, ABCD3, ZNF198, FGF9, FLT3, FLT1, BRCA2, FKHR, TPT1, LCP1, RB1, DACH, PIBF1, KLF12, GPC5, GPC5 (2), GPC6, ABCC4, RAP2A, FGF14, ERCC5, EFNB2, ING1, TFDP1, GAS6, D13S327, TEP1, MMP14, BCL2L2, FKHL1, MBIP, HNF3A, SSTR1, PNN (DRS), CDKN3 RBBP1, DAAM1, MNAT1, HIF1A, ESR2, MAX, EIF2S1, RAD51L1, IGHG1, HSXIAPAF1, MN1, and TSPY genes were found (Table 30). Heterozygous deletion of these genes was detected as frequently as 40-89% of the Myeloma cell lines tested.

  Furthermore, EPS15, PGR, YAP1, cIAP1, MMP7, MMP1, DYNEIN, ETS1, FLI1, IGHG1, and TSPY genes were found as genes with a ratio of 0.25 or less, that is, homozygote deletion (Table 31). ). These heterozygous and homozygous gene groups may cause canceration due to a marked decrease in the expression level.

  In particular, homozygous deletion genes are tumor suppressor genes, and deletion of these gene groups plays an important role in inducing canceration.

[Example 11] Comprehensive analysis of cancer-related genes in Anaplastic thyroid cell line (thyroid cancer cell line) using MCG cancer array Amplified and deleted using Anatomical thyroid cell line using "MCG cancer array" The genes that have been analyzed. As a result of investigating genes amplified with a Ratio of 1.32 or more, genes p63, TERT, DOC2, PPP1A, E2F1, and AIB1 were found (Table 32). Amplification of these genes was detected in 6-11 of the 14 Anaplastic thyroid cell lines tested and was frequent.

  The MET, NRG1, MYC, PVT1, YAP1, and cIAP1 genes were found as genes in which the ratio was 4 or more in the Anaplastic thyroid cell line, that is, 4 times or more amplification was detected compared to the normal cell gene ( Table 33).

  Next, in the Anaplastic thyroid cell line, the ratio decreased to 0.75 or less, that is, as genes judged to be heterozygotes, CTNNB1, BAIAP1, ABCE1, ESR1, N33, LZTS1, DMBT1, EGF9, PMP22, SMAD4-2, ADRBK2 genes Were found (Table 34). Heterozygous deletion of these genes was detected in 4-9 of the 14 Analastic thyroid cell lines tested and was frequent.

Furthermore, MLLT3 and CDKN2A (p16) genes were found as genes with a ratio of 0.25 or less, that is, homozygote deletion (Table 34). These heterozygous and homozygous gene groups may cause canceration due to a marked decrease in the expression level.

  In particular, homozygous deletion genes are tumor suppressor genes, and deletion of these gene groups plays an important role in inducing canceration.

[Example 12] Inhibition of growth of squamous cell sarcoma by treatment with Sendai virus incorporating CDKN2A (p16) gene and treatment of tumor-bearing nude mice
The CDKN2A (p16) gene is ligated downstream of the Survivin promoter and incorporated into the Sendai virus vector. Packaging of the obtained viral DNA is performed to produce a recombinant virus. Recombinant virus is purified using Discontinuous iodixanol gradient centrifugation and a heparin agarose column. Inoculate squamous cell carcinoma in a 96-well tissue culture plate at a concentration of 5 × 10 ³ cells / well and culture in a CO ₂ incubator at 37 ° C. for 1 day. Each well was infected with 100 moi of purified recombinant Sendai virus, and after 72 hours of culture, the level of cell proliferation was determined by 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide assay. It was measured. The measurement was performed according to the instructions using a commercially available kit (Promega, Tokyo). As a result, significant growth inhibition was found in cells infected with the virus incorporating CDKN2A (p16) gene. When Western blot analysis was performed on the cell extract, CDKN2A (p16) protein was not detected in control Sendai virus-infected cells, whereas a clear CDKN2A (p16) protein band was detected from the cell extract sample. was detected. This result indicates that the infection of Sendai virus with CDKN2A (p16) gene is suppressed, and the growth of cancer cells is suppressed. It can be used to reduce cancer or to suppress micrometastasis. Means.

Next, inoculate squamous cell carcinoma using nude mice, simultaneously infect with 3 × 10 ¹¹ moi of purified recombinant Sendai virus, monitor cancer growth inhibition, and increase the survival time of the mice, ie, this gene An increase in response rate due to treatment can be expected.

  Based on the results, clinical trials of gene therapy for human cancer patients can be planned.

[Example 13] Growth inhibition of small cell lung cancer cells by SKP2 gene antisense oligonucleotide The chromosome 5p13 region is amplified in small cell lung cancer cells. As a result of examining ACC-LC-5, ACC-LC-172, Lu-130, and Lu-134 among small cell lung cancer cell lines, the CDH6, PC4, and SKP2 genes present in the 5p13 region were Southern blotted. The method confirmed significant amplification at the chromosomal level and found a significant increase compared to normal cells where expression of those genes was analyzed using Northern blotting (FIG. 3). As a result of culturing these cells in the presence of SKP2 antisense oligonucleotide, the amount of SKP2 mRNA is significantly reduced, and the cell growth is suppressed to a level of 25% compared with the control sense-added cells. It was done. The added SKP2 antisense oligonucleotide reached the maximum inhibition at a concentration of 200 nM-1 μM. The time course of cell proliferation decreased to a level of 25% after 4 days (FIG. 4).

  FIG. 4 (A) shows RNA from cells treated with Untreated, treated with transfection reagent only (Oligofectamine), transfected with SKP2 antisense oligonucleotide (AS), and transfected with SKP2 sense oligonucleotide (SC). The result of preparing and analyzing the expression level of SKP2 mRNA by Northern blotting is shown. It can be seen that SKP2 mRNA expression is significantly reduced by AS treatment.

  FIG. 4 (B) shows the results of examining the inhibition of growth of small cell lung cancer cells by SKP2 antisense oligonucleotide. The vertical axis represents cell growth, and the growth of untreated cells was taken as 100% and expressed as a relative%. Proliferation was examined by adding SKP2 sense oligonucleotide and SKP2 antisense oligonucleotide in the range of 0-1000 nM.

  FIG. 4 (C) shows the time course of small cell lung cancer cell proliferation by SKP2 antisense oligonucleotide. The vertical axis is the same as (B). The horizontal axis represents the number of culture days after the addition of SKP2 sense oligonucleotide and SKP2 antisense oligonucleotide.

  FIG. 5 is a drawing in which apoptosis induction of ACC-LC172 cells by addition of SKP2 antisense oligonucleotide was examined. In FIG. 5, it was found that in ACC-LC172 cells, addition of SKP2 antisense oligonucleotide increased the number of cells killed by apoptosis from 3% to 25% untreated (FIG. 5B). Furthermore, apoptosis was confirmed by morphological observation and flow cytometry analysis.

  FIG. 5A shows micrographs of control cells (Of), cells transfected with SKP2 antisense oligonucleotide (AS), and cells transfected with SKP2 sense oligonucleotide (SC). AS showed typical results for cells that induced apoptosis. FIG. 5B shows the percentage of cells in which apoptosis occurred (vertical axis). Of, AS, and SC have the same meaning as in FIG. 5A. It can be seen that apoptosis was remarkably induced by treatment with SKP2 antisense oligonucleotide. FIG. 5C shows the results of flow cytometry of ACC-LC172 cells treated with AS, again showing typical results of apoptosis.

  These results prove that SKP2 antisense oligonucleotides specifically inhibit the growth of cancer cells, and SKP2 antisense oligonucleotides can be used as cancer therapeutic agents. .

[Example 14] Decrease in expression due to methylation of DBCCR1 gene in non-small cell lung cancer cells When eight strains of non-small cell lung cancer cells were examined for deletion of DBCCR1 gene by genomic PCR, homozygous deletion was shown. There was only one cell and the remaining 7 cells had 2 copies. Next, when the expression of the DBCCR1 gene was analyzed using the RT-PCR method, the expression of DBCCR1 gene was not detected at all in 8 cells. When the degree of methylation was examined by the method, it was found that all were methylated. Therefore, it was found that the expression of the DBCCR1 gene was suppressed in many non-small cell lung cancer cells by methylation.

It is explanatory drawing of a normal cell (normal diploid cell) genome analysis using a MCG cancer array. It is the figure which showed the result of the cancer cell genome analysis using the MCG cancer array. It is drawing which shows the result of having examined about the amplification level of SKP2 gene, PC4 gene, and CDH6 gene which exist in 5p13 in a small cell lung cancer cell, and the expression level of SKP2 gene and PC4 gene. It is the figure which showed the result of having examined the growth inhibition of the small cell lung cancer cell by SKP2 antisense oligonucleotide addition. It is a figure which shows the result of having examined the apoptosis induction of the ACC-LC172 cell by SKP2 antisense oligonucleotide addition.

Claims (49)

Group consisting of the following genes in the specimen:
MUC1 gene, PRCC gene, EIF4G gene, THPO gene, TERT gene, DAB2 gene, EGFR gene, ELN gene, MUC3 gene, MYC gene, PVT1 gene, FCA gene, PTPN1 gene, Livin gene, MYCN gene, ELK3 gene, BCL2L2 gene , HNF3A gene, ERBB2 gene, CGI-147 gene, ESP15 gene, ARHGEF2 gene, ABCC5 gene, EIF4G1 gene, CDH12 gene, PC4 gene, SKP2 gene, ZNF131 gene, RAD52 gene, WNT3 gene, CDC27 gene, COL1A1 gene, ABCC3 gene , PPM1D gene, ITGB4 gene, BIRC5 gene, SHGC-103396 gene, BCL2L1 gene, RBL1 gene, SRC gene, TGIF2 gene, MYBL2 gene, BIRC7 gene, ARAF1 gene, CUL4B gene, CTAG1 gene, TRIM33 gene, MYCL1 gene, RLF gene , CCNE1 gene, PCTK1 gene, ETV5 gene, CDC10 gene, IGFBP1 gene, TCRG gene, MYCLK1 gene, TAX1BP1 gene, IL6 gene, PMS2 gene, MET residue Gene, SMOH gene, BRAF gene, CDK5 gene, AR gene, MCF2 gene, MAGEA2 gene, CTAG gene, ALX gene, PMF1 gene, NTRK1 gene, ERV5 gene, MUC4 gene, PDGFRA gene, IGFBP7 gene, CDH10 gene, E2F3 gene , TPMT gene, TFAP2A gene, EEF1E1 gene, RREB1 gene, CDK6 gene, PRIM1 gene, GLI gene, CDK4 gene, FUS gene, CYLD gene, GRB2 gene, TGFβR3 gene, PAX3 gene, MLL gene, FKHR gene, FOLR1 gene, PLUNC (LUNX) gene, E2F1 gene, TNFRSF5 gene, NCOA3 gene, ELMO2 gene, NCOA3 (AIB1) gene, PRex1 gene, BCAS1 gene, ZNF217 gene, STK6 (BTAK) gene, SDC1 gene, DNMT3A gene, MLH1 gene, CTNNB1 gene, CCK gene, EGR2 gene, KSAM (FGFR2) gene, PKY (HIPK3) gene, LMO2 gene, CD44 gene, KRAS gene, KRAG (SSPN) gene, CYP1A1 gene, IQGAP1 gene, FURIN ( PACE) gene, PPARBP gene, KRAG gene, KRAS2 gene, PTHLH gene, BCLX gene, DEK gene, Livin-2 gene, TFAP2C gene, TNFRSF6B gene, HCK gene, CDC2L1 gene, GLI3 gene, PPP1A gene, SUPT5H gene, AKT2 gene , TRRAP gene, Smurf1 gene, PDAP1 gene, MIA gene SERPINE1 gene, VGF gene, HRAS gene, BCL3 gene, LUNX gene, AIB1 gene, NCOA gene, SSX4 gene, SSX1 gene, MCL1 gene, CCND3 gene, FLT3 gene, DOC2 gene , BAI1 gene, PSCA gene, MLZE gene, RECQL4 gene, BCL1 gene, FGF4 gene, Survivin gene, BCL9 gene, AF1Q gene, DAP3 gene, BRAL1 gene, TRK gene, CRP gene, ATF6 gene, PBX1 gene, ABL2 gene, LAMC2 Gene, TP gene, ABL gene, CAN gene, NCOR1 gene, ZNF287 gene, D17S128 gene, BCL2 gene, FVT1 gene, PI5 gene, p63 gene, NRG1 Genes, YAP1 genes and cIAP1 genes;
A method for detecting cancer, comprising detecting canceration of the sample using amplification of one or more genomic genes selected from The cancer is lung squamous cell carcinoma, lung adenocarcinoma, or large cell carcinoma, and consists of the following genes:
MUC1 gene, PRCC gene, EIF4G gene, THPO gene, TERT gene, DAB2 gene, EGFR gene, ELN gene, MUC3 gene, MYC gene, PVT1 gene, FCA gene, PTPN1 gene, Livin gene, MYCN gene, ELK3 gene, BCL2L2 gene , HNF3A gene, ERBB2 gene and CGI-147 gene;
The cancer detection method according to claim 1, wherein canceration of a specimen is detected using one or more kinds of genomic genes selected from the above as an index of amplification of 1.5 times or more compared to normal cells. The cancer is lung squamous cell carcinoma, lung adenocarcinoma, or large cell carcinoma, and consists of the following genes:
MYCN gene, MYC gene, PVT1 gene, ELK3 gene, BCL2L2 gene, HNF3A gene, ERBB2 gene and CGI-147 gene;
The method for detecting cancer according to claim 1, wherein canceration of a specimen is detected using one or more kinds of genomic genes selected from the above as an index compared to normal cells. The cancer is non-small cell lung cancer and is composed of the following genes:
ESP15 gene, MUC1 gene, ARHGEF2 gene, PRCC gene, ABCC5 gene, EIF4G1 gene, CDH12 gene, PC4 gene, SKP2 gene, DAB2 gene, ZNF131 gene, RAD52 gene, WNT3 gene, CDC27 gene, COL1A1 gene, ABCC3 gene, PPM1D gene , ITGB4 gene, BIRC5 gene, SHGC-103396 gene, BCL2L1 gene, RBL1 gene, SRC gene, TGIF2 gene, MYBL2 gene, BIRC7 gene, ARAF1 gene, CUL4B gene, CTAG1 gene, TRIM33 gene, MYCL1 gene, RLF gene, MYC gene CCNE1 gene and PCTK1 gene;
The method for detecting cancer according to claim 1, wherein canceration of a specimen is detected by using amplification of 1.5 times or more as an index compared to normal cells for one or more genomic genes selected from The cancer is non-small cell lung cancer and is composed of the following genes: TRIM33 gene, MYCL1 gene, RLF gene, MYC gene, CCNE1 gene and PCTK1 gene;
The method for detecting cancer according to claim 1, wherein canceration of a specimen is detected using one or more kinds of genomic genes selected from the above as an index compared to normal cells. The cancer is a malignant glioma and consists of the following genes:
EIF4G gene, ETV5 gene, EGFR gene, CDC10 gene, IGFBP1 gene, TCRG gene, MYCLK1 gene, TAX1BP1 gene, IL6 gene, PMS2 gene, MUC3 gene, MET gene, SMOH gene, BRAF gene, CDK5 gene, AR gene, CUL4B gene , MCF2 gene, MAGEA2 gene, CTAG gene, ALX gene, MUC1 gene, ARHGEF2 gene, PMF1 gene, NTRK1 gene, ERV5 gene, MUC4 gene, PDGFRA gene, IGFBP7 gene, PC4 gene, SKP2 gene, DAB2 gene, CDH10 gene, CDH12 Gene, TERT gene, E2F3 gene, TPMT gene, TFAP2A gene, EEF1E1 gene, RREB1 gene, CDK6 gene, PRIM1 gene, GLI gene, CDK4 gene, FUS gene, CYLD gene and GRB2 gene;
The cancer detection method according to claim 1, wherein canceration of a specimen is detected using one or more kinds of genomic genes selected from the above as an index of amplification of 1.5 times or more compared to normal cells. The cancer is a malignant glioma and consists of the following genes:
ALX gene, MUC1 gene, ARHGEF2 gene, PMF1 gene, NTRK1 gene, ERV5 gene, MUC4 gene, PDGFRA gene, IGFBP7 gene, PC4 gene, SKP2 gene, DAB2 gene, CDH10 gene, CDH12 gene, TERT gene, E2F3 gene, TPMT gene , TFAP2A gene, EEF1E1 gene, RREB1 gene, EGFR gene, PMS2 gene, CDK6 gene, PRIM1 gene, GLI gene, CDK4 gene, FUS gene, CYLD gene and GRB2 gene;
The method for detecting cancer according to claim 1, wherein canceration of a specimen is detected using one or more kinds of genomic genes selected from the above as an index compared to normal cells. When the cancer is a neuroblastoma and one or more genomic genes selected from the group consisting of MYCL1, CDH10, and MYC genes, the amplification of the specimen is 1.5 times or more compared to normal cells. The method for detecting cancer according to claim 1, wherein canceration is detected. The cancer is a neuroblastoma (Neuroblastoma) and canceration of a specimen is detected using amplification of one or more genomic genes selected from the group consisting of MYCN gene, CDK4 gene and PPM1D gene as an index. The method for detecting cancer as described. The cancer is rhabdomyosarcoma and consists of the following genes:
TGFβR3 gene, PAX3 gene, MLL gene, and FKHR gene;
The cancer detection method according to claim 1, wherein canceration of a specimen is detected using one or more kinds of genomic genes selected from the above as an index of amplification of 1.5 times or more compared to normal cells. The method for detecting cancer according to claim 1, wherein the cancer is rhabdomyosarcoma, and the canceration of the specimen is detected using CDK4 genomic gene as an indicator with an amplification of 4 times or more compared to normal cells. . The cancer is gastric cancer and comprises the following genes:
PVT1 gene, MYC gene, FOLR1 gene, PLUNC (LUNX) gene, E2F1 gene, TGIF2 gene, TNFRSF5 gene, NCOA3 gene, ELMO2 gene, MYBL2 gene, NCOA3 (AIB1) gene, PTPN1 gene, PRex1 gene, BCAS1 gene, ZNF217 gene , STK6 (BTAK) gene, CUL4B gene, MCF2 gene, CTAG gene, SDC1 gene, DNMT3A gene, MLH1 gene, CTNNB1 gene, CCK gene, ZNF131 gene, CDK6 gene, MET gene, PVT1 gene, EGR2 gene, KSAM (FGFR2) Gene, PKY (HIPK3) gene, LMO2 gene, CD44 gene, KRAS gene, KRAG (SSPN) gene, CYP1A1 gene, IQGAP1 gene, FURIN (PACE) gene, PPARBP gene, ERBB2 gene, CCNE1 gene and MYBL2 gene;
The method for detecting cancer according to claim 1, wherein canceration of a specimen is detected using amplification of 1.5 times or more as an index compared to normal cells for one or more genomic genes selected from The cancer is gastric cancer and comprises the following genes:
SDC1 gene, DNMT3A gene, MLH1 gene, CTNNB1 gene, CCK gene, ZNF131 gene, CDK6 gene, MET gene, MYC gene, PVT1 gene, EGR2 gene, KSAM (FGFR2) gene, PKY (HIPK3) gene, LMO2 gene, CD44 gene , KRAS gene, KRAG (SSPN) gene, CYP1A1 gene, IQGAP1 gene, FURIN (PACE) gene, PPARBP gene, ERBB2 gene, CCNE1 gene and MYBL2 gene;
The method for detecting cancer according to claim 1, wherein canceration of a specimen is detected using one or more kinds of genomic genes selected from the above as an index compared to normal cells. The cancer is pancreatic cancer and comprises the following genes:
KRAG gene, PTPN1 gene, KRAS2 gene, PTHLH gene, BCLX gene, DEK gene, IGFBP1 gene, MYC gene, Livin-2 gene, PVT1 gene, PRex1 gene, BCAS1 gene, TFAP2C gene, EGFR gene, TGIF2 gene, TNFRSF5 gene, TNFRSF6B gene, EIF4G gene, PMS2 gene, HCK gene, MYBL2 gene, ELMO2 gene, PCTK1 gene, CDC2L1 gene, CDC10 gene, TCRG gene, GLI3 gene, PPP1A gene, ZNF217 gene, SRC gene, SUPT5H gene, AKT2 gene, TRRAP gene , Smurf1 gene, PDAP1 gene, PVT1 gene and MIA gene;
The cancer detection method according to claim 1, wherein canceration of a specimen is detected using one or more kinds of genomic genes selected from the above as an index of amplification of 1.5 times or more compared to normal cells. The cancer is pancreatic cancer and comprises the following genes:
SUPT5H gene, AKT2 gene, TRRAP gene, Smurf1 gene, PDAP1 gene, MYC gene, PVT1 gene, KRAS2 gene, KRAG gene and MIA gene;
The method for detecting cancer according to claim 1, wherein canceration of a specimen is detected using one or more kinds of genomic genes selected from the above as an index compared to normal cells. The cancer is colorectal cancer and consists of the following genes:
ELN gene, SERPINE1 gene, VGF gene, MUC3 gene, MYC gene, PVT1 gene, HRAS gene, BCL3 gene, BCLX gene, LUNX gene, E2F1 gene, TGIF2 gene, HCK gene, AIB1 gene, PTPN1 gene, NCOA gene, TNFRSF6B gene , SSX4 gene, SSX1 gene, ARAF1 gene, CUL4B gene, CTAG gene, MAGEA2 gene, MCL1 gene, CCND3 gene and FLT3 gene;
The cancer detection method according to claim 1, wherein canceration of a specimen is detected using one or more kinds of genomic genes selected from the above as an index of amplification of 1.5 times or more compared to normal cells. The cancer is colorectal cancer and consists of the following genes:
MCL1 gene, CCND3 gene, MYC gene, PVT1 gene and FLT3 gene;
The cancer detection method according to claim 1, wherein canceration of a specimen is detected using an amplification of 4 or more times as an index compared to normal cells for one or more genomic genes selected from The cancer is cholangiocarcinoma and consists of the following genes:
ZNF131 gene, DOC2 gene, DAB2 gene, PC4 gene, SKP2 gene, CDH10 gene, CDH12 gene, TERT gene, CDK5 gene, BAI1 gene, PSCA gene, MLZE gene, RECQL4 gene, BCL1 gene, FGF4 gene, ITGB4 gene, Survivin gene , SRC gene, PTPN1 gene, PCTK1 gene, CTAG gene;
The cancer detection method according to claim 1, wherein canceration of a specimen is detected using one or more kinds of genomic genes selected from the above as an index of amplification of 1.5 times or more compared to normal cells. The cancer is a myeloid species and consists of the following genes:
BCL9 gene, MCL1 gene, AF1Q gene, MUC1 gene, DAP3 gene, ARHGEF2 gene, PMF1 gene, BRAL1 gene, PRCC gene, TRK gene, NTRK1 gene, CRP gene, ATF6 gene, PBX1 gene, ABL2 gene, LAMC2 gene, TP gene , ABL gene, CAN gene, KRAS2 gene, KRAG gene, PTHLH gene, NCOR1 gene, ZNF287 gene, D17S128 gene, BCL2 gene, FVT1 gene and PI5 gene;
The method for detecting cancer according to claim 1, wherein canceration of a specimen is detected using amplification of 1.5 times or more as an index compared to normal cells for one or more genomic genes selected from The cancer is a myeloid species and consists of the following genes:
ABL gene, CAN gene, KRAS2 gene, KRAG gene, PTHLH gene, NCOR1 gene, ZNF287 gene, D17S128 gene, BCL2 gene, FVT1 gene and PI5 gene;
The method for detecting cancer according to claim 1, wherein canceration of a specimen is detected using one or more kinds of genomic genes selected from the above as an index compared to normal cells. The cancer is thyroid cancer and comprises the following genes:
p63 gene, TERT gene, DOC2 gene, PPP1A gene, E2F1 gene and AIB1 gene;
The cancer detection method according to claim 1, wherein canceration of a specimen is detected using one or more kinds of genomic genes selected from the above as an index of amplification of 1.5 times or more compared to normal cells. The group consisting of the following genes:
MET gene, NRG1 gene, MYC gene, PVT1 gene, YAP1 gene and cIAP1 gene;
The cancer detection method according to claim 1, wherein canceration of a specimen is detected using one or more kinds of genomic genes selected from the above as an index of amplification of 1.5 times or more compared to normal cells. Group consisting of the following genes in the specimen:
BAIAP1 gene, LZTS1 gene, AAC1 gene, BLK gene, MTAP gene, CDKN2A (p16) gene, GPC5 gene, SNRPN gene, SAMD4 gene, DCC gene, ADRBK2 gene, DBCCR1 gene, RIZ gene, CCK gene, UBE2E1 gene, THRB gene , RARB gene, TGFβR2 gene, MLH1 gene, CTNNB1 gene, VIPR1 gene, ZNF35 gene, TGM4 gene, TDGF1 gene, PTPRG gene, FHIT gene, MITF gene, LOC151987 gene, EIF4E gene, NFκB gene, CCNA gene, PGRMC2 gene, VEGC Gene, MSH3 gene, RASA1 gene, TPT1 gene, RB1 gene, AATF gene, NR2F1 gene, AFP gene, EGF5 gene, ABCG2 gene, BMI1 gene, PCDH15 gene, PGR gene, FGF9 gene, ZNF198 gene, FLT1 gene, FLT3 gene, BRCA2 gene, KLF12 gene, PIBF1 gene, HNF3A gene, MBIP gene, FKHL1 gene, CDKN2A (p16) gene, N33 gene, TEK gene, MLLT3 gene, JAK 2 genes, GASC1 gene, D9S913 gene, SMAD4 gene, MADH2 gene, MADH7 (SMAD7) gene, MALT1 gene, GRP gene, BCL2 gene, FVT1 gene, SERPINB (PI5) gene, CTDP1 gene, SMAD4-2 gene, D8S504 gene, NAT2 gene, TNFRSF10B gene, MAP3K7 gene, DLC1 gene, stSG42796 gene, LPL gene, NRG1 gene, SCCA1 gene, SCCA2 gene, NKX3A gene, SMAD7 gene, MLL1 gene, PI5 gene, Casp3 gene, SSXT gene, VIM gene, stSG27915 gene , RH68621 gene, SHGC-145820 gene, EEF1E1 gene, ESR1 gene, MLLT3 gene, DEC1 gene, CDH23 gene, ETK1 gene, VIP gene, IGHG1 gene, PMP22 gene, MAFG gene, SHGC-145820 gene, FGF2 gene, ST5 gene, CALCA gene, FKHR gene, CXADR gene, TGFβR3 gene, ABCD3 gene, LCP1 gene, DACH gene, GPC5 (2) gene, GPC6 gene, ABCC4 gene, RAP2A gene , FGF14 gene, ERCC5 gene, EFNB2 gene, ING1 gene, TFDP1 gene, GAS6 gene, D13S327 gene, TEP1 gene, MMP14 gene, BCL2L2 gene, SSTR1 gene, PNN (DRS) gene, CDKN3 gene, RBBP1 gene, DAAM1 gene, MNAT1 Gene, HIF1A gene, ESR2 gene, MAX gene, EIF2S1 gene, RAD51L1 gene, HSXIAPAF1 gene, MN1 gene, TSPY gene, EPS15 gene, YAP1 gene, cIAP1 gene, MMP7 gene, MMP1 gene, DYNEIN gene, ETS1 gene, FLI1 gene, IGHG1 gene, TSPY gene, ABCE1 gene, DMBT1 gene and EGF9 gene;
Detecting canceration of the specimen using as an index the deletion of one or more genomic genes selected from
Cancer detection method. The cancer is lung squamous cell carcinoma, lung adenocarcinoma, or large cell carcinoma, and consists of the following genes:
BAIAP1 gene, LZTS1 gene, AAC1 gene, BLK gene, MTAP gene, CDKN2A (p16) gene, GPC5 gene, SNRPN gene, SAMD4 gene, DCC gene, ADRBK2 gene, DBCCR1 gene and RIZ gene;
24. The method for detecting cancer according to claim 23, wherein canceration of the specimen is detected using a heterozygote deletion of one or more genomic genes selected from The cancer is lung squamous cell carcinoma, lung adenocarcinoma, or large cell carcinoma, and consists of the following genes:
CDKN2A (p16) gene, MTAP gene, DBCCR1 gene and RIZ gene;
The cancer detection method according to claim 23, wherein canceration of a specimen is detected using a homozygous deletion of one or more genomic genes selected from The cancer is non-small cell lung cancer and is composed of the following genes:
CCK gene, UBE2E1 gene, THRB gene, RARB gene, TGFβR2 gene, MLH1 gene, CTNNB1 gene, VIPR1 gene, ZNF35 gene, TGM4 gene, TDGF1 gene, PTPRG gene, FHIT gene, BAIAP1 gene, MITF gene, LOC151987 gene, EIF4E gene , NFκB gene, CCNA gene, PGRMC2 gene, VEGC gene, MSH3 gene, RASA1 gene, TPT1 gene, RB1 gene, AATF gene, ADRBK2 gene and NR2F1 gene;
24. The method for detecting cancer according to claim 23, wherein canceration of the specimen is detected using a heterozygote deletion of one or more genomic genes selected from The cancer detection method according to claim 23, wherein the cancer is non-small cell lung cancer, and canceration of the specimen is detected using a homozygous deletion of the NR2F1 genomic gene as an index. The cancer is a malignant glioma and consists of the following genes:
AFP gene, EGF5 gene, ABCG2 gene, NFκB gene, MTAP gene, BMI1 gene, PCDH15 gene, PGR gene, FGF9 gene, ZNF198 gene, FLT1 gene, FLT3 gene, BRCA2 gene, RB1 gene, KLF12 gene, PIBF1 gene, HNF3A gene , MBIP gene, FKHL1 gene, MTAP gene and CDKN2A (p16) gene;
24. The method for detecting cancer according to claim 23, wherein canceration of the specimen is detected using a heterozygote deletion of one or more genomic genes selected from The method for detecting cancer according to claim 23, wherein the cancer is malignant glioma, and canceration of the specimen is detected using a homozygous deletion of the MTAP gene and / or CDKN2A (p16) genomic gene as an index. The cancer is gastric cancer and comprises the following genes:
MTAP gene, DCC gene, N33 gene, AAC1 gene, GRP gene, TEK gene, D8S504 gene, NAT2 gene, LZTS1 gene, TNFRSF10B gene, D9S913 gene, GASC1 gene, FVT1 gene, MAP3K7 gene, DLC1 gene, MALT1 gene, stSG42796 gene , BAIAP1 gene, BLK gene, LPL gene, NRG1 gene, MLLT3 gene, MADH2 gene, SCCA1 gene, SCCA2 gene, NKX3A gene, SMAD7 gene, MLL1 gene, PI5 gene, Casp3 gene, SSXT gene, BCL2 gene, JAK2 gene, PTPRG Gene, VIM gene, stSG27915 gene, RH68621 gene, CTDP1 gene, SHGC-145820 gene, EEF1E1 gene, ESR1 gene, KLF12 gene, CDKN2A (p16) gene, N33 gene, DEC1 gene, CDH23 gene and SMAD4-2 gene;
24. The method for detecting cancer according to claim 23, wherein canceration of the specimen is detected using a heterozygote deletion of one or more genomic genes selected from The cancer is gastric cancer and comprises the following genes:
MTAP gene, CDKN2A (p16) gene, TEK gene, RB1 gene and SNRPN gene;
The cancer detection method according to claim 23, wherein canceration of a specimen is detected using a homozygous deletion of one or more genomic genes selected from The cancer is pancreatic cancer and comprises the following genes:
MTAP gene, DCC gene, N33 gene, AAC1 gene, GRP gene, TEK gene, D8S504 gene, NAT2 gene, LZTS1 gene, TNFRSF10B gene, D9S913 gene, GASC1 gene, FVT1 gene, MAP3K7 gene, DLC1 gene, MALT1 gene, stSG42796 gene , BAIAP1 gene, BLK gene, LPL gene, NRG1 gene, MLLT3 gene, MADH2 gene, SCCA1 gene, SCCA2 gene, NKX3A gene, SMAD7 gene, MLL1 gene, PI5 gene, Casp3 gene, SSXT gene, BCL2 gene, JAK2 gene, PTPRG Gene, VIM gene, stSG27915 gene, RH68621 gene, CTDP1 gene, SHGC-145820 gene, EEF1E1 gene, ESR1 gene, KLF12 gene gene, CDKN2A (p16) gene, DEC1 gene, CDH23 gene and SMAD4-2 gene;
24. The method for detecting cancer according to claim 23, wherein canceration of the specimen is detected using a heterozygote deletion of one or more genomic genes selected from The cancer is pancreatic cancer and comprises the following genes:
CDKN2A (p16) gene, MTAP gene, N33 gene, MLLT3 gene, TEK gene, DEC1 gene, CDH23 gene and SMAD4-2 gene;
The cancer detection method according to claim 23, wherein canceration of a specimen is detected using a homozygous deletion of one or more genomic genes selected from The cancer is colorectal cancer and consists of the following genes:
ETK1 gene, MITF gene, PTPRG gene, FHIT gene, RARB gene, VEGFC gene, MAP3K7 gene, VIP gene, N33 gene, D8S504 gene, PCDH15 gene, IGHG1 gene, PMP22 gene, MAFG gene, SSXT gene, MADH2 gene, DCC gene , SMAD4-2 gene, GRP gene, CTDP1 gene and SHGC-145820 gene;
24. The method for detecting cancer according to claim 23, wherein canceration of the specimen is detected using a heterozygote deletion of one or more genomic genes selected from The cancer is bile duct cancer and consists of the following genes:
BAIAP1 gene, PTPRG gene, TDGF1 gene, EIF4E gene, NFκB gene, CCNA gene, FGF2 gene, NKX3A gene, N33 gene, LZTS1 gene, LPL gene, NRG1 gene, DLC1 gene, BLK gene, AAC1 gene, NAT2 gene, D8S504 gene , MTAP gene, JAK2 gene, ST5 gene, CALCA gene, FLT3 gene, FLT1 gene, FKHR gene and CXADR gene;
24. The method for detecting cancer according to claim 23, wherein canceration of the specimen is detected using a heterozygote deletion of one or more genomic genes selected from 24. The method for detecting cancer according to claim 23, wherein the cancer is cholangiocarcinoma and canceration of the specimen is detected using a homozygous deletion of the CXADR genomic gene as an index. The cancer is myeloma and consists of the following genes:
TGFβR3 gene, ABCD3 gene, ZNF198 gene, FGF9 gene, FLT3 gene, FLT1 gene, BRCA2 gene, FKHR gene, TPT1 gene, LCP1 gene, RB1 gene, DACH gene, PIBF1 gene, KLF12 gene, GPC5 gene, GPC5 (2) gene , GPC6 gene, ABCC4 gene, RAP2A gene, FGF14 gene, ERCC5 gene, EFNB2 gene, ING1 gene, TFDP1 gene, GAS6 gene, D13S327 gene, TEP1 gene, MMP14 gene, BCL2L2 gene, FKHL1 gene, MBIP gene, HNF3A gene, SSTR1 Gene, PNN (DRS) gene, CDKN3 gene, RBBP1 gene, DAAM1 gene, MNAT1 gene, HIF1A gene, ESR2 gene, MAX gene, EIF2S1 gene, RAD51L1 gene, IGHG1 gene, HSXIAPAF1 gene, MN1 gene, TSPY gene, EPS15 gene, PGR gene, YAP1 gene, cIAP1 gene, MMP7 gene, MMP1 gene, DYNEIN gene, ETS1 gene and FLI1 gene;
24. The method for detecting cancer according to claim 23, wherein canceration of the specimen is detected using a heterozygote deletion of one or more genomic genes selected from The cancer is myeloma and consists of the following genes:
EPS15 gene, PGR gene, YAP1 gene, cIAP1 gene, MMP7 gene, MMP1 gene, DYNEIN gene, ETS1 gene, FLI1 gene, IGHG1 gene and TSPY gene;
The cancer detection method according to claim 23, wherein canceration of a specimen is detected using a homozygous deletion of one or more genomic genes selected from The cancer is thyroid cancer and comprises the following genes:
CTNNB1, BAIAP1, ABCE1, ESR1, N33, LZTS1, DMBT1, EGF9, PMP22, SMAD4-2, ADRBK2 gene, MLLT3 gene and CDKN2A (p16) gene;
24. The method for detecting cancer according to claim 23, wherein canceration of the specimen is detected using a heterozygote deletion of one or more genomic genes selected from The method for detecting cancer according to claim 23, wherein the cancer is thyroid cancer, and canceration of the specimen is detected using a homozygous deletion of the MLLT3 genomic gene and / or CDKN2A (p16) genomic gene as an index. A method for detecting cancer, wherein non-small cell lung cancer is detected by detecting suppression of DBCCR1 gene expression in a specimen. The detection method according to any one of claims 1 to 41, wherein the detection method is performed by a CGH method, a DNA chip method, a quantitative PCR method, or a real-time RT-PCR method. 43. The DNA fixing substrate according to claim 42, wherein the detection method is carried out by a CGH method or a DNA chip method, and the plurality of types of DNA to be fixed on the detection substrate are genomic DNA, cDNA, or synthetic oligonucleotide. The detection method is CGH, and the plurality of types of DNA to be fixed on the detection substrate are genomic DNA, and the genomic DNA is a gene amplification product of BAC DNA, YAC DNA, or PAC DNA. 43. A DNA fixing substrate according to 43. A method for suppressing a cancer cell by introducing into the cancer cell a gene whose gene deletion is associated with canceration of the cell. 46. The method for suppressing cancer cells according to claim 45, wherein the cancer to be suppressed and the genes whose gene deletion is associated with canceration of the cells are the following groups 1 to 9.
1. Esophageal cancer: LRP1B gene;
2. Non-small cell lung cancer: one or more genes selected from the group consisting of MTAP gene, CDKN2A (p16) gene, DBCCR1 gene and 1RIZ gene;
3. Small cell lung cancer: NR2F1 gene;
4). Malignant glioma: MTAP gene and / or CDKN2A (p16) gene;
5). Gastric cancer: one or more genes selected from the group consisting of MTAP gene, CDKN2A (p16) gene, TEK gene, RB1 gene and SNRPN gene;
6). 6. Pancreatic cancer: one or more genes selected from the group consisting of N13 gene, MTAP gene, CDKN2A (p16) gene, TEK gene, MLLT3 gene, DEC-1 gene, CDH23 gene and SMAD4-2 gene; Cholangiocarcinoma: CXADR gene;
8). Myeloma: one or more genes selected from the group consisting of EPS15 gene, PGR gene, cIAP1 gene, MMP1 gene, DYNEIN gene, YAP1 gene, MMP7 gene, ETS1 gene, FLI1 gene, IGHG1 gene and TSPY gene;
9. Thyroid cancer: CDKN2A (p16) gene and / or MLLT3 gene; A method of inhibiting cancer cells by causing nucleic acid that antagonizes a transcription product of a gene associated with canceration of the cells to act on the cancer cells. 48. The method for suppressing cancer cells according to claim 47, wherein the nucleic acid that antagonizes the transcript of the gene is a small interference RNA for the mRNA that is the transcript or an antisense oligonucleotide thereof. 49. The method of suppressing cancer cells according to claim 47 or 48, wherein the cancer to be suppressed and the genes whose gene amplification is related to canceration of the cells are the following groups 1 to 8.
1. Small cell lung cancer: one or more genes selected from the group consisting of MYCN gene, MYC gene, PVT1 gene, ELK3 gene, BCL2L2 gene, HNF3A gene, ERBB2 gene and CGI-147 gene;
2. Non-small cell lung cancer: one or more genes selected from the group consisting of TRIM33 gene, MYCL1 gene, RLF gene, MYC gene, CCNE1 gene and PCTK1 gene;
3. Malignant glioma: ALX gene, ARHGEF2 gene, PC4 gene, SKP2 gene, DAB2 gene, PMF1 gene, NTRK1 gene, ERV5 gene, CDH10 gene, CDH12 gene, TERT gene, MUC4 gene, PDGFRA gene, IGFBP7 gene, E2F3 gene, TPMT gene One or more genes selected from the group consisting of TFAP2A gene, EEF1E1 gene, RREB1 gene, EGFR gene, GLI gene, CDK4 gene, FUS gene, PMS2 gene, CDK6 gene, PRIM1 gene, CYLD gene and GRB2 gene;
4). Gastric cancer: SDC1 gene, DNMT3A gene, MLH1 gene, PKY gene, LMO2 gene, CD44 gene, CTNNB1 gene, CCK gene, KRAS gene, KRAG gene, CYP1A1 gene, CDK6 gene, MET gene, MYC gene, IQGAP1 gene, FURIN gene, One or more genes selected from the group consisting of PPARBP gene, PVT1 gene, EGR2 gene, EGFR2 gene, ERBB2 gene, CCNE1 gene and MYBL2 gene;
5). Pancreatic cancer: one or more genes selected from the group consisting of SUPT5H gene, TRRAP gene, PVT1 gene, KRAS2 gene, KRAG gene, Smurf1 gene, PDAP1 gene, MYC gene and MIA gene;
6). Colorectal cancer: one or more genes selected from the group consisting of MCL1 gene, CCND3 gene, MYC gene, PVT1 gene and FLT3 gene;
7). Myeloma: one or more genes selected from the group consisting of ABL gene, CAN gene, KRAS2 gene, ZNF285 gene, D17S128 gene, BCL2 gene, KRAG gene, PTHLH gene, NCOR1 gene, FVT1 gene and PI5 gene;
8). Thyroid cancer: one or more genes selected from the group consisting of MET gene, NRG1 gene, MYC gene, PVT1 gene, YAP1 gene and cIAP1 gene;