2002 年度実績報告書

体系的遺伝子異常解析に基づく乳癌・甲状腺癌の遺伝子診断法の開発

研究課題

研究課題/領域番号	12217141
研究機関	日本医科大学
研究代表者	江見充日本医科大学, 老人病研究所, 教授 (90221118)
キーワード	乳癌 / 甲状腺癌 / 遺伝子診断 / マイクロアレイ / 技術移転
研究概要	昨年度詳細に実験条件の検討を行ったcDNAマイクロアレイ解析法によって、乳癌・甲状腺癌の新鮮凍結保存癌部および非癌部検体より高純度RNAを抽出し、T7 RNAポリメラーゼによるRNA増幅を2回行い、約10000倍のポリ(A)RNAを得た。癌部および非癌部由来のRNAを、2種類の蛍光色素にて標識してcDNAプローブを合成し、約2万5千種類のcDNAを高密度にスポットしたマイクロアレイ上でハイブリダイゼーションを行った。マイクロアレイ上のシグナルデータのコンピュータ解析を行い、約2万5千種類の遺伝子の癌組織における発現プロファイル解析を行った。2色の蛍光色素量の正規化に用いるため各々のマイクロアレイ上にはそれぞれ384種のハウスキーピング遺伝子をスポットし、これらのハウスキーピング遺伝子に基づいて正規化を行った。数理統計解析の適正化・手法向上の結果、信頼性の高い実験結果を得ることが可能となった。1)エストロゲン受容体陰性乳癌における5年以内再発死亡群と5年以上健存群の比較、2)T2乳癌における同様の比較、3)n1β乳癌における同様の比較、また、4)n0乳癌における5年以内再発群と5年以上健存群間の比較、5)甲状腺乳頭癌における再発群と健存群の比較、6)甲状腺未分化癌の特性の検討、の各々の課題についてcDNAマイクロアレイを用いた体系的遺伝子発現解析を行ってい、各々の群において発現異常を呈するDVL-1,XBP-1などの遺伝子を同定した。さらに、これら遺伝子についてRT-PCR法にて多検体で確認、抗原・抗体作製による臨床診断法の開発を進めている。

研究成果
(23件)

すべてその他

すべて文献書誌 (23件)

[文献書誌] Nagai, H.: "Combined Hypermethylation and Chromosome Loss Associated with Inactivation of SSI-1/SOCS-1/JAB Gene in Human Hepatocellular Carcinomas"Cancer Lett. 186(1). 59-65 (2002)
[文献書誌] Nagahata, T.: "Correlation of Allelic Losses and Clinicopathological Factors in 504 Primary Breast Cancers"Breast Cancer. 9(3). 208-215 (2002)
[文献書誌] Iwasaki, H.: "Accuracy of Genotyping for Single Nucleotide Polymorphism Typing Method involving hybridization of Short Allele-Specific Oligonucleotides"DNA Res.. 9(2). 59-62 (2002)
[文献書誌] Harada, H.: "Genomic structure and eight novel exonic polymorphisms of human N-cadherin gene"J. Hum. Genet.. 47(6). 330-332 (2002)
[文献書誌] Yoshida, S.: "Head-to-head Juxtaposition of Fas-Associated Phosphatase-1(FAP-1) and c-Jun NH_2-terminal Kinase 3(JNK3) genes : genomic structure and five polymorphisms of FAP-1 gene"J. Hum. Genet.. 47(11). 614-619 (2002)
[文献書誌] Shimizu, M.: "Universal Fluorescent Labeling (UFL) Method for Automated Microsatellite Analysis"DNA Res.. 9(5). 173-178 (2002)
[文献書誌] Harada. H.: "Down-regulation of a Novel Gene, DRLM, in human Liver Malignancy from 4q22 That Encodes an NAP-like Protein"Gene. 296(1-2). 171-177 (2002)
[文献書誌] Tsuneizumi, M.: "A highly polymorphic CA repeat marker at the EBAG9/RCAS1 locus on 8q23 that detected frequent multiplication in breast cancer"Annals Hum. Bio.. 29(4). 457-460 (2002)
[文献書誌] Watanabe, T.: "Differentially regulated genes as putative targets of amplifications at 20qin ovarian cancers"Jpn. J. Cancer. Res.. 93(10). 1114-1122 (2002)
[文献書誌] Shinohara, Y.: "Three TNFα single nucleotide polymorphisms in the Japanese population"An. Hum. Bio.. 29(5). 579-583 (2002)
[文献書誌] Nakata, T.: "Differential Expression of Multiple Isoforms of the ELKS mRNAs Involved in a Papillary Thyroid Carcinoma"Genes, Chrom. Cancer. 35(1). 30-37 (2002)
[文献書誌] Harada, H.: "Genomic Structure of the Human NLK(nemo-like kinase) Gene and Analysis of its Promoter Region"Gene. 285(1-2). 175-182 (2002)
[文献書誌] Nakajima, T.: "Nucleotide Diversity and Haplotype Structure of the Human Angiotensinogen Gene in Two Populations"Am. J. Hum. Genet.. 70(1). 108-123 (2002)
[文献書誌] Tsuneizumi, M.: "Association of Allelic loss at 8p22 with Poor prognosis among Breast Cancers Cases treated with high-dose adjuvant chemotherapy"Cancer Lett.. 180(1). 75-82 (2002)
[文献書誌] Fujiwara, H.: "Association of common missense changes in ELAC2 (HPC2) with prostate cancer in a Japanese case-control series"J. Hum. Genet.. 47(12). 641-648 (2002)
[文献書誌] Nagahata, T.: "Molecular Diagnosis for Breast Cancer"Jpn. Med. Assoc. J.. 45. 265-260 (2002)
[文献書誌] Yasui, K.: "TFDP1, CUL4A, and CDC16 Identified as Targets for Amplification at 13q34 in Hepatocellular Carcinomas"Hepatology. 35(6). 1476-1484 (2002)
[文献書誌] Nagai, H.: "Hypermethylation Associated with Inactivation of the SOCS-1 Gene, a JAK/STAT inhibitor, in Human Hepatoblastomas"J. Hum. Genet.. 48(2). 65-69 (2002)
[文献書誌] Nagai, H.: "DNA Alterations during Multi-step Development of Human Hepatocellular Carcinomas Revealed by Laser Capture Microdissection"Hepatology Res.. (in press). (2002)
[文献書誌] Nagai, H.: "Down-regulation in Human Cancers of DRHC, a Novel Helicase-like Gene from 17q25.1 That Inhibit Cell Growth"Cancer Lett.. (in press). (2003)
[文献書誌] 岩崎公典: "ゲノム研究各論〜DNAチップ、マイクロアレイ〜"分子心血管病. 3(3). 109-114 (2002)
[文献書誌] 永井尚生: "肝硬変における遺伝子発現異常の解析"消化器科. 34(1). 64-68 (2002)
[文献書誌] 永井尚生: "癌の遺伝要因の解明と現状"医学のあゆみ. 202(10). 869-876 (2002)

2002 年度 実績報告書

体系的遺伝子異常解析に基づく乳癌・甲状腺癌の遺伝子診断法の開発

研究代表者

江見 充 日本医科大学, 老人病研究所, 教授 (90221118)

研究成果

[文献書誌] Nagai, H.: "Combined Hypermethylation and Chromosome Loss Associated with Inactivation of SSI-1/SOCS-1/JAB Gene in Human Hepatocellular Carcinomas"Cancer Lett. 186(1). 59-65 (2002)

[文献書誌] Nagahata, T.: "Correlation of Allelic Losses and Clinicopathological Factors in 504 Primary Breast Cancers"Breast Cancer. 9(3). 208-215 (2002)

[文献書誌] Iwasaki, H.: "Accuracy of Genotyping for Single Nucleotide Polymorphism Typing Method involving hybridization of Short Allele-Specific Oligonucleotides"DNA Res.. 9(2). 59-62 (2002)

[文献書誌] Harada, H.: "Genomic structure and eight novel exonic polymorphisms of human N-cadherin gene"J. Hum. Genet.. 47(6). 330-332 (2002)

[文献書誌] Yoshida, S.: "Head-to-head Juxtaposition of Fas-Associated Phosphatase-1(FAP-1) and c-Jun NH_2-terminal Kinase 3(JNK3) genes : genomic structure and five polymorphisms of FAP-1 gene"J. Hum. Genet.. 47(11). 614-619 (2002)

[文献書誌] Shimizu, M.: "Universal Fluorescent Labeling (UFL) Method for Automated Microsatellite Analysis"DNA Res.. 9(5). 173-178 (2002)

[文献書誌] Harada. H.: "Down-regulation of a Novel Gene, DRLM, in human Liver Malignancy from 4q22 That Encodes an NAP-like Protein"Gene. 296(1-2). 171-177 (2002)

[文献書誌] Tsuneizumi, M.: "A highly polymorphic CA repeat marker at the EBAG9/RCAS1 locus on 8q23 that detected frequent multiplication in breast cancer"Annals Hum. Bio.. 29(4). 457-460 (2002)

[文献書誌] Watanabe, T.: "Differentially regulated genes as putative targets of amplifications at 20qin ovarian cancers"Jpn. J. Cancer. Res.. 93(10). 1114-1122 (2002)

[文献書誌] Shinohara, Y.: "Three TNFα single nucleotide polymorphisms in the Japanese population"An. Hum. Bio.. 29(5). 579-583 (2002)

[文献書誌] Nakata, T.: "Differential Expression of Multiple Isoforms of the ELKS mRNAs Involved in a Papillary Thyroid Carcinoma"Genes, Chrom. Cancer. 35(1). 30-37 (2002)

[文献書誌] Harada, H.: "Genomic Structure of the Human NLK(nemo-like kinase) Gene and Analysis of its Promoter Region"Gene. 285(1-2). 175-182 (2002)

[文献書誌] Nakajima, T.: "Nucleotide Diversity and Haplotype Structure of the Human Angiotensinogen Gene in Two Populations"Am. J. Hum. Genet.. 70(1). 108-123 (2002)

[文献書誌] Tsuneizumi, M.: "Association of Allelic loss at 8p22 with Poor prognosis among Breast Cancers Cases treated with high-dose adjuvant chemotherapy"Cancer Lett.. 180(1). 75-82 (2002)

[文献書誌] Fujiwara, H.: "Association of common missense changes in ELAC2 (HPC2) with prostate cancer in a Japanese case-control series"J. Hum. Genet.. 47(12). 641-648 (2002)

[文献書誌] Nagahata, T.: "Molecular Diagnosis for Breast Cancer"Jpn. Med. Assoc. J.. 45. 265-260 (2002)

[文献書誌] Yasui, K.: "TFDP1, CUL4A, and CDC16 Identified as Targets for Amplification at 13q34 in Hepatocellular Carcinomas"Hepatology. 35(6). 1476-1484 (2002)

[文献書誌] Nagai, H.: "Hypermethylation Associated with Inactivation of the SOCS-1 Gene, a JAK/STAT inhibitor, in Human Hepatoblastomas"J. Hum. Genet.. 48(2). 65-69 (2002)

[文献書誌] Nagai, H.: "DNA Alterations during Multi-step Development of Human Hepatocellular Carcinomas Revealed by Laser Capture Microdissection"Hepatology Res.. (in press). (2002)

[文献書誌] Nagai, H.: "Down-regulation in Human Cancers of DRHC, a Novel Helicase-like Gene from 17q25.1 That Inhibit Cell Growth"Cancer Lett.. (in press). (2003)

[文献書誌] 岩崎公典: "ゲノム研究各論〜DNAチップ、マイクロアレイ〜"分子心血管病. 3(3). 109-114 (2002)

[文献書誌] 永井尚生: "肝硬変における遺伝子発現異常の解析"消化器科. 34(1). 64-68 (2002)

[文献書誌] 永井尚生: "癌の遺伝要因の解明と現状"医学のあゆみ. 202(10). 869-876 (2002)

2002 年度実績報告書

江見充日本医科大学, 老人病研究所, 教授 (90221118)