2000 Fiscal Year Annual Research Report

がん原物質の活性化および解毒化酵素の機能と発がんリスク

Research Project

Project/Area Number	12213002
Research Category	Grant-in-Aid for Scientific Research on Priority Areas (C)
Research Institution	Hokkaido University
Principal Investigator	鎌滝哲也北海道大学, 大学院・薬学研究科, 教授 (00009177)
Co-Investigator(Kenkyū-buntansha)	十川和博東北大学, 大学院・理学研究科, 助教授 (80175421)
Keywords	チトクロームP450 / 代謝的活性化 / サルモネラ菌 / CYP1A2 / 誘導的発現 / Ah受容体 / CYP2A6 / 発がんリスクの個体差
Research Abstract	化学発がんのイニシエーション段階では,生体内の異物代謝酵素による化学物質の代謝的活性化と解毒化反応が関わり,その両者のバランスが個体の発がんリスクを左右すると予想される.本研究では,1)環境中の変異原物質がヒトの異物代謝酵素で実際に活性化されるか否か2)ヒトの異物代謝酵素活性の個体差が発がんリスクの個体差と直接関連するか否か3)これら酵素の発現量を制御する発現制御機構の未解明の問題の解決を目的としている.本年度は,1)について,がん原物質を代謝的に活性化するヒトの異物代謝酵素チトクロームP450分子種の1つであるCYP1B1を発現する二種類のサルモネラ菌(TA1538およびYG7108)株の構築に成功し,それぞれ,Trp-P-1,煙草特異的ニトロソアミンであるNNKなどの変異原・がん原物質の高感度な検出系として有用であることを明らかにした.また,発がん物質の解毒化にかかわるヒトUDP-グルクロン酸転移酵素(UGT)の1分子種(UGT1A1)について,活性を有する形でサルモネラ菌に発現させることにはじめて成功した.さらに,2)については,日本人を用いたケース・コントロール研究で3種類のがんを検討したところ,CYP2A6の遺伝的多型が肺がんの感受性と関連することを明らかにし,CYP2A6遺伝子の欠損者では,肺がんリスクが有意に低い(オッズ比0.25,95%信頼区間,0.08-0.83)ことを明らかにした.最後に,3)については,焼け焦げに含まれるがん原性ヘテロサイクリックアミン類の代謝的活性化に重要な役割を果たしているCYP1A2について,申請者らはその発現を制御する領域を見出した.今年度は,この領域に直接結合するタンパウ質を常用されるクロマトグラフィーおよび,エンハンサー配列を持つオリゴヌクレオチドによるDNAアフィニティークロマトグラフィーを用いることにより精製することに成功した.得られた蛋白質の分子量は約60kDaであることが明らかとなった.

Research Products

(20 results)

All Other

All Publications (20 results)

[Publications] Noritaka Ariyoshi et al.: "A Single Nucleotide Polymorphism of CYP2B6 Found in Japanese Enhances Catalytic Activity by Autoactivation."Biochemical Biophysical Research Communications. (In press). (2001)
[Publications] Noritaka Ariyoshi et al.: "A Novel Single Nucleotide Polymorphism Altering Stability and Activity of CYP2A6."Biochemical Biophysical Research Communications. (In press). (2001)
[Publications] Takeshi Ozeki et al.: "Cooperative regulation of the transcription of human dihydrodiol dehydrogenase (DD)4/AKR1C4 gene by hepatocyte neclear factor (HNF)-4α/γ and HNF-1α."Biochemical Journal. (In press). (2001)
[Publications] Noritaka Ariyoshi et al.: "Pharmacogenetics on cancer chemotherapy : from basic studies."Japanese Journal of Clinical Pharmacology and Therapeutics. (In press). (2001)
[Publications] Taisei Mushiroda et al.: "Accumulation of the 1-methyl-4-phenylpyridinium ion (MPP+) in suncus (Suncus murinus) brain : Implication for flavin-containing monooxygenase activity in brain microvessels."Chemical Research in Toxicology. (In press). (2001)
[Publications] Loriot,M-A et al.: "Genetic polymorphism of cytochrome P450 2A6 in a case control study on lung cancer in a French population."Pharmacogenetics. 11. 39-44 (2001)
[Publications] Noritaka Ariyoshi et al.: "Studies on the genetic polymorphism of CYP2A6."Drug Metabolism and Disposition. 15. 57-61 (2000)
[Publications] Hirotaka Kushida et al.: "Development of a Salmonella tester strain sensitive to promutagenic N-nitrosamines : expression of recombinant CYP2A6 and human NADPH-cytochrome P450 reductase in S.typhimurium YG7108."Mutation Research. 471. 135-143 (2000)
[Publications] Takashi Satoh et al.: "Studies on the interactions between drugs and estrogen. Analytical method for prediction system of gynecomastia induced by drugs on the inhibitory metabolism of estradiol using Escherichia coli co-expressing human CYP3A4 with human NADPH-cytochrome P450 reductase."Analytical Biochemistry. 286. 179-186 (2000)
[Publications] Noritaka Ariyoshi et al.: "Structural characterization of a new variant of the CYP2A6 gene (CYP2A6*1B) appparently diagnosed as heterozygotes of CYP2A6*1A and CYP2A6*4C."Pharmacogenetics. 10. 678-693 (2000)
[Publications] Taisei Mushiroda et al.: "The house musk shrew (Suncus murinus): a unique animal with extremely low level of expression of mRNA for CYP3A and flavin-containing monooxygenase."Comparative Biochemistry & Physiology. 126. 225-234 (2000)
[Publications] Hirotaka Kushida et al.: "Metabolic activation of N-alkylnitrosamines in genetically engineered Salmonella typhimurium expressing CYP2E1 or CYP2A6 together with human NADPH-cytochrome P450 reductase."Carcinogenesis. 21. 1227-1232 (2000)
[Publications] Eri Inoue et al.: "Development of bacterial expression system with high yield of CYP3A7, a human fetus-specific form of Cytochrome P450."Biochemical Biophysical Research Communications. 269. 623-627 (2000)
[Publications] Taisei Mushiroda et al.: "A high level expression of CYP2A in the lung of the Suncus (Suncus murinus) and its role in the Activation of 4-(methylntirosamino)-1-(3-pyridyl)-1-butanone (NNK)."Biochemical Biophysical Research Communications. 260. 393-396 (2000)
[Publications] Ken-ichi Fujita et al.: "Establishment of Salmonella strain expressing catalytically active human UDP-glucuronosyltransferase 1A1 (UGT1A1)"Life Science. 66. 1955-1967 (2000)
[Publications] Noritaka Ariyoshi et al.: "Handbook of Experimental Pharmacology"Springer-VerlagBerlin (Not determined). (2001)
[Publications] 有吉範高: "月刊薬事2001年臨時増刊号"じほう(Not determined). (2001)
[Publications] 鎌滝哲也: "薬物療法学"南江堂(Not determined). (2001)
[Publications] 有吉範高: "月刊薬事3月臨時増刊号"じほう. 834 (2000)
[Publications] 鎌滝哲也: "薬物代謝学"東京化学同人. 282 (2000)

2000 Fiscal Year Annual Research Report

がん原物質の活性化および解毒化酵素の機能と発がんリスク

Principal Investigator

鎌滝 哲也 北海道大学, 大学院・薬学研究科, 教授 (00009177)

Research Products

[Publications] Noritaka Ariyoshi et al.: "A Single Nucleotide Polymorphism of CYP2B6 Found in Japanese Enhances Catalytic Activity by Autoactivation."Biochemical Biophysical Research Communications. (In press). (2001)

[Publications] Noritaka Ariyoshi et al.: "A Novel Single Nucleotide Polymorphism Altering Stability and Activity of CYP2A6."Biochemical Biophysical Research Communications. (In press). (2001)

[Publications] Takeshi Ozeki et al.: "Cooperative regulation of the transcription of human dihydrodiol dehydrogenase (DD)4/AKR1C4 gene by hepatocyte neclear factor (HNF)-4α/γ and HNF-1α."Biochemical Journal. (In press). (2001)

[Publications] Noritaka Ariyoshi et al.: "Pharmacogenetics on cancer chemotherapy : from basic studies."Japanese Journal of Clinical Pharmacology and Therapeutics. (In press). (2001)

[Publications] Taisei Mushiroda et al.: "Accumulation of the 1-methyl-4-phenylpyridinium ion (MPP+) in suncus (Suncus murinus) brain : Implication for flavin-containing monooxygenase activity in brain microvessels."Chemical Research in Toxicology. (In press). (2001)

[Publications] Loriot,M-A et al.: "Genetic polymorphism of cytochrome P450 2A6 in a case control study on lung cancer in a French population."Pharmacogenetics. 11. 39-44 (2001)

[Publications] Noritaka Ariyoshi et al.: "Studies on the genetic polymorphism of CYP2A6."Drug Metabolism and Disposition. 15. 57-61 (2000)

[Publications] Hirotaka Kushida et al.: "Development of a Salmonella tester strain sensitive to promutagenic N-nitrosamines : expression of recombinant CYP2A6 and human NADPH-cytochrome P450 reductase in S.typhimurium YG7108."Mutation Research. 471. 135-143 (2000)

[Publications] Noritaka Ariyoshi et al.: "Structural characterization of a new variant of the CYP2A6 gene (CYP2A6*1B) appparently diagnosed as heterozygotes of CYP2A6*1A and CYP2A6*4C."Pharmacogenetics. 10. 678-693 (2000)

[Publications] Taisei Mushiroda et al.: "The house musk shrew (Suncus murinus): a unique animal with extremely low level of expression of mRNA for CYP3A and flavin-containing monooxygenase."Comparative Biochemistry & Physiology. 126. 225-234 (2000)

[Publications] Hirotaka Kushida et al.: "Metabolic activation of N-alkylnitrosamines in genetically engineered Salmonella typhimurium expressing CYP2E1 or CYP2A6 together with human NADPH-cytochrome P450 reductase."Carcinogenesis. 21. 1227-1232 (2000)

[Publications] Eri Inoue et al.: "Development of bacterial expression system with high yield of CYP3A7, a human fetus-specific form of Cytochrome P450."Biochemical Biophysical Research Communications. 269. 623-627 (2000)

[Publications] Taisei Mushiroda et al.: "A high level expression of CYP2A in the lung of the Suncus (Suncus murinus) and its role in the Activation of 4-(methylntirosamino)-1-(3-pyridyl)-1-butanone (NNK)."Biochemical Biophysical Research Communications. 260. 393-396 (2000)

[Publications] Ken-ichi Fujita et al.: "Establishment of Salmonella strain expressing catalytically active human UDP-glucuronosyltransferase 1A1 (UGT1A1)"Life Science. 66. 1955-1967 (2000)

[Publications] Noritaka Ariyoshi et al.: "Handbook of Experimental Pharmacology"Springer-VerlagBerlin (Not determined). (2001)

[Publications] 有吉範高: "月刊薬事2001年臨時増刊号"じほう(Not determined). (2001)

[Publications] 鎌滝哲也: "薬物療法学"南江堂(Not determined). (2001)

[Publications] 有吉範高: "月刊薬事3月臨時増刊号"じほう. 834 (2000)

[Publications] 鎌滝哲也: "薬物代謝学"東京化学同人. 282 (2000)

鎌滝哲也北海道大学, 大学院・薬学研究科, 教授 (00009177)

[Publications] Noritaka Ariyoshi et al.: "Structural characterization of a new variant of the CYP2A6 gene (CYP2A61B) appparently diagnosed as heterozygotes of CYP2A61A and CYP2A6*4C."Pharmacogenetics. 10. 678-693 (2000)