FISH法による癌悪性度判定法の開発

• Project/Area Number: 09256204
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas (A)
• Allocation Type: Single-year Grants
• Research Institution: Tokyo Medical and Dental University (1998-1999); The University of Tokyo (1997)
• Principal Investigator: 稲澤 譲治 東京医科歯科大学, 難治疾患研究所, 教授 (30193551)
• Project Period (FY): 1999
• Project Status: Completed (Fiscal Year 1999)
• Budget Amount: ¥40,000,000 (Direct Cost: ¥40,000,000); Fiscal Year 1999: ¥20,000,000 (Direct Cost: ¥20,000,000); Fiscal Year 1998: ¥10,000,000 (Direct Cost: ¥10,000,000); Fiscal Year 1997: ¥10,000,000 (Direct Cost: ¥10,000,000)
• Keywords: 癌 / 遺伝子増幅 / 癌遺伝子 / 癌抑制遺伝子 / FISH / CGH / 診断 / ヒトゲノム / 腫瘍 / 染色体異常 / SKY / ゲノム / 遺伝子診断

Research Abstract

各種病型の腫瘍を対象にゲノム変化をCGH法で全染色体領域にわたり網羅的にスクリーニングし、悪性度に関与する増幅領域を探索した。見つけだされた新規増幅領域を標的に増幅内遺伝子の同定・単離と機能解析を進めた。胃癌(GC),食道扁平上皮癌(ESC),腎細胞(RCC),卵巣癌(OCC),膵臓癌(PC),肝細胞癌(HCC)、悪性線維性組織球腫(MFH)等の固形腫瘍と成人T細胞性白血病(ATL)においてCGHによる染色体コピー数異常の解析を終了した、(計130株以上)。その結果、新規の増幅領域として複数の細胞株で、1q32,3q36-37,4q12,5P15,7q31,8q22-23,9p23,11q12-14,14q13-21,15q26,17q23-24,18p13等を見出したGC株で検出した15q26増幅領域内に、遺伝子増幅により発現亢進する既知遺伝子IQGAP1と新規遺伝子を同定した。未分化型GC株に比較的高頻度に検出した新規11p13増幅領域内に既知遺伝子ならびに新規遺伝子の3種類をそれぞれ同定した。ESC株において1q32増幅領域内でATF3とCENPFが増幅/発現亢進していることを明らかにした。9p23-24増幅の共通増幅領域から新規の癌関連遺伝子GASC1を単離した。本遺伝子は全長4239bp(1056aaをコード)でPHD,PXドメインを有す。本増幅領域は乳癌の罹病性遺伝子の存在領域とも一致する。RCC株の非乳頭型腎細胞癌において5q31-33領域(D5S624-D5S673)の優先的コピー数増加を明らかにした。本領域内にはPDGFRB,CSF1R,FGF1,IL9,CDC25,CDC25Cなどの増殖関連遺伝子が密に存在する。OCC、HCC細胞株で高頻度検出する14q13-21増幅、またESCの3q36-27増幅において、遺伝子増幅により活性化を受けている既知遺伝子を同定した。悪性線維性組織球腫(Malignant Fibrous Histiocytoma;MFH)で見出した8p23.1増幅領域より新規の増幅遺伝子MASL1を単離した。

Research Products

• [Publications] Yang, Z-Q., Inazawa J. et al.: "Molecular cytogenetic analysis of 17 renal-cancer cell lines : increased copy number at 5q31-33 in cell lines from nonpapillary carcinomas"Jpn J Cancer Res. 91. 156-163 (2000)
• [Publications] Sugimoto, N., Inazawa J. et al.: "The human caspase-activated DNase (CAD) gene : genomic structure, exonic single-nucleotide polymorphisms, and a highly polymorphic dinucleotide repeat at the CAD"J Hum Genet. 44. 408-411 (1999)
• [Publications] Sakabe, T., Inazawa J. et al.: "Identification of a Novel Gene, MASL1, Within an Amplicon at 8p23.1 Detected in Malignant Fibrous Histiocytomas (MFH) by Comparative Genomic Hybridization"Cancer Res. 51. 511-515 (1999)
• [Publications] Sakakura, C., Inazawa J. et al.: "Gains, losses, and amplifications of genomic materials in primary gastric cancers analyzed by comparative genomic hybridization"Genes Chromosomes Cancer. 24. 299-305 (1999)
• [Publications] Shinoyama, T., Inazawa J. et al.: "Comparative genomic hybridization of squamous cell carcinoma of the esophagus : The possible involvement of the DP1 gene in the 13q34 amplicon"Genes Chromosomes Cancer. 24. 337-344 (1999)
• [Publications] Sakakura, C., Inazawa J. et al.: "Chromosomal aberrations in human hepatocellular carcinomas associated with hepatitis C virus detected by comparative genomic hybridization"Br J Cancer. 80. 2034-2039 (1999)
• [Publications] Ishino S,Inazawa J et al.: "Loss of material from chromosome arm 1p during malignant progression of meningioma revealed by fluorescent in situ hybridization." Cancer. 83. 360-366 (1998)
• [Publications] Mukae N,Inazawa J,Nagata S et al.: "Molecular cloning and characterization of human caspase-activated DNase" Proc Natl Acad Sci USA. 95. 9123-9128 (1998)
• [Publications] Mori T,Inazawa J,et al.: "Cloning and characterization of a novel Rad-family gene,Rab36,within the region at 22q11.2 that Is homozygously deleted in malignant rhabdoid tumors" Biochem Biophys Res Commun. 254. 594-600 (1999)
• [Publications] Shinomiya T,Inazawa J et al.: "Comparative genomic hybridization of squamous cell carcinoma of the esophagus : The possible involvement of the DP1 gene in the 13q34 amplicon" Genes Chromosomes Cancer. (in press).
• [Publications] Sakakura C,Inazawa J,et al.: "Gains,losses,and amplifications of genomic materals in primary gastric cancers analyzed by comparative genomic hybridization." Genes Chromosomes Cancer. (in press).
• [Publications] Sakabe T,Inazawa J et al.: "Identification of a Novel Gene,MASL1,Within an Amplicon at 8p23.1 Detected in Malignant Fibrous Histiocytomas(MFH)by Comparative Genomic Hybridization" Cancer Res. (in press).
• [Publications] 稲澤譲治 他: "ゲノム医学の新しい展開" 講談社ライフサイエンティフィク, 126頁 (1998)
• [Publications] Houri,T.et al: "Chromosomal translocation,t(1;11)(q12;p15),in an extragonadal immature teratoma." Cancer Genet.Cytogenet.97. 79-80 (1997)
• [Publications] Shimotake,T.et al.: "Germline mutation of the RET proto-oncogene in children with total intestinal aganglionosis." J Pediatr Surg. 32. 498-500 (1997)
• [Publications] Saito,H.et al.: "Frequent association of alternative splicing of NER,a nuclear hormone receptor gene in cancer tissues." Oncogene. 14. 617-621 (1997)
• [Publications] Tsuda,H., et al.: "Detection of numerical and structural alterations and fusion of chromosomes 16 and 1 in low-grade papillary breast carcinoma by fluorescence in situ hybridization." Am J Pathol. 151. 1027-1034 (1997)
• [Publications] Murata,Het al.: "Ploidy analysis in paraffin-embedded malignant fibrous histiocytoma by DNA cytofluorometry and fluorescence in situ hybridization." Cancer Lett.118. 123-128 (1997)
• [Publications] Ishino,S.et al.: "Loss pf material from chromosome arm of 1p during malignant progression of meningioma revealed by fluorescent in situ hybridization(FISH)." Cancer. (in press.).