2000 Fiscal Year Annual Research Report

活性酸素によるDNA損傷とその防御

Research Project

Project/Area Number	12213137
Research Category	Grant-in-Aid for Scientific Research on Priority Areas (C)
Research Institution	University of Occupational and Environmental Health, Japan
Principal Investigator	葛西宏産業医科大学, 産業生態科学研究所, 教授 (40152615)
Co-Investigator(Kenkyū-buntansha)	平野雄産業医科大学, 産業生態科学研究所, 助手 (40258629) 紙谷浩之北海道大学, 大学院・薬学研究科, 助教授 (10204629)
Keywords	2-OH-dATP / 2-OH-ATP / MTH1 / 8-OH-dG / クロロゲン酸 / 4NQO / 3'-methyl-4-DAB
Research Abstract	活性酸素によるDNA損傷とその防御に関して下記のような、大腸菌を用いた変異誘発に関する基礎的研究、酵素基質特異性に関する生化学的研究、また発がんリスク評価に関連して、動物臓器DNA、ヒト試料の酸化ストレスのマーカーの分析などを行った。 1.酸化損傷ヌクレオシド三リン酸、2-ヒドロキシデオキシアデノシン(2-OH-dATP)は大腸菌DNAポリメラーゼIIIにより高頻度でグアニンに対合して取り込まれ、主にG→T変異を起こした。 2.リボヌクレオチド2-OH-ATPはヒトMTH1蛋白により効率よく分解され様々な酸化損傷ヌクレオチドの中で最も低いKm値(最も高い親和性)を示した。 3.野菜、果物中の主要抗酸化成分クロロゲン酸は4NQOによるラット舌発がんモデルにおいてDNA中の8-ヒドロキシデオキシグアノシン(8-OH-dG)の生成を抑制した。 4.栄養欠乏食およびエタノールの長期投与によりラット食道DNA中の8-OH-dGおよびその修復活性は上昇し酸化ストレスの関与が示された。 5.アゾ色素系肝発がん物質3'-methyl-4-dimethylaminoazobenzenの投与によりマウスおよびラットの肝臓中の8-OH-dGおよびその修復活性が高まったことから肝発がんにおける活性酸素の関与が示された。 6.尿中8-OH-dG測定法を開発し、某企業従業員約300人の尿を分析し、ライフスタイルとの関係を調べたところ、適度の運動(<5時間/週)は8-OH-dG値を低下させること等がわかった。今後は、2-OH-Adeを細胞レベルで検出し、発がんにおけるこのDNA損傷の意義を研究したい。また8-OH-dGについては更に分析を進め癌予防のためのライフスタイルの開発に役立てたい。

Research Products

(11 results)

All Other

All Publications (11 results)

[Publications] Kamiya,H., et al.: "2-Hydroxy-dATP is incorporated opposite G by Escherichia coli DNA polymerase III resulting in high mutagenicity."Nucleic Acids Res.. 28. 1640-1646 (2000)
[Publications] Kasai,H., et al.: "Action of Chlorogenic acid in vegetables and fruits as an inhibitor of 8-hydroxydeoxyguanosine formation in vitro and in a rat carcinogenesis model."Food and Chemical Toxycology. 38. 467-471 (2000)
[Publications] Hirano,T., et al.: "Analysis of oxidative DNA damage and its repair activity in the livers of 3'-methyl-4-dimethylaminoazobenzene-treated rodents."Jpn.J.Cancer Res.. 91. 681-685 (2000)
[Publications] Kamiya,H., et al.: "Two DNA polymerases of Escherichia coli display distinct misinsertion specificities for 2-hydroxy-dATP during DNA syntheses."Biochemistry. 39. 9508-9513 (2000)
[Publications] Murata-Kamiya,N., et al.: "Methylglyoxal induces G : C to C : G and G : C to T : A transversions in the supF gene on a shuttle vector plasmid replicated in mammalian cells."Mutation Res.. 468. 173-182 (2000)
[Publications] Kamiya,H., et al.: "2-Hydroxyadenine formation by reactive oxygen species and mutagenic effects."Recent Res.Devel.Biochem.. 2. 41-50 (2000)
[Publications] Asami,S., et al.: "Increase in 8-hydroxyguanine and its repair activity in the esophagi of rats given long-term ethanol and nutrition-deficient diet."Jpn.J.Cancer Res.. 91. 973-978 (2000)
[Publications] Kamiya,H., et al.: "2-Hydroxyadenine in DNA is a very poor substrate of the Escherichia coli MutY protein."J.Radiat.Res.. 41. 349-354 (2000)
[Publications] Fujikawa,K., et al.: "Human MTH1 protein hydrolyzes the oxidized ribonucleotide, 2-hydroxy-ATP."Nucleic Acids Res.. 29. 449-454 (2001)
[Publications] Kasai,H., et al.: "Life style and urinary 8-hydroxydeoxyguanosine, a marker of oxidative DNA damage : Effects of exercise, working conditions, meat intake, body mass index, and smoking."Jpn.J.Cancer Res.. 92. 9-15 (2001)
[Publications] Kasai,H., et al.: "Free Radicals in Chemistry, Biology and Medicine. (eds.Yoshikawa,T,et al)"Analysis of 8-hydroxydeoxyguanosine in cellular DNA and urine as a marker or oxidative stress.. 8 (2000)

2000 Fiscal Year Annual Research Report

活性酸素によるDNA損傷とその防御

Principal Investigator

葛西 宏 産業医科大学, 産業生態科学研究所, 教授 (40152615)

Research Products

[Publications] Kamiya,H., et al.: "2-Hydroxy-dATP is incorporated opposite G by Escherichia coli DNA polymerase III resulting in high mutagenicity."Nucleic Acids Res.. 28. 1640-1646 (2000)

[Publications] Kasai,H., et al.: "Action of Chlorogenic acid in vegetables and fruits as an inhibitor of 8-hydroxydeoxyguanosine formation in vitro and in a rat carcinogenesis model."Food and Chemical Toxycology. 38. 467-471 (2000)

[Publications] Hirano,T., et al.: "Analysis of oxidative DNA damage and its repair activity in the livers of 3'-methyl-4-dimethylaminoazobenzene-treated rodents."Jpn.J.Cancer Res.. 91. 681-685 (2000)

[Publications] Kamiya,H., et al.: "Two DNA polymerases of Escherichia coli display distinct misinsertion specificities for 2-hydroxy-dATP during DNA syntheses."Biochemistry. 39. 9508-9513 (2000)

[Publications] Murata-Kamiya,N., et al.: "Methylglyoxal induces G : C to C : G and G : C to T : A transversions in the supF gene on a shuttle vector plasmid replicated in mammalian cells."Mutation Res.. 468. 173-182 (2000)

[Publications] Kamiya,H., et al.: "2-Hydroxyadenine formation by reactive oxygen species and mutagenic effects."Recent Res.Devel.Biochem.. 2. 41-50 (2000)

[Publications] Asami,S., et al.: "Increase in 8-hydroxyguanine and its repair activity in the esophagi of rats given long-term ethanol and nutrition-deficient diet."Jpn.J.Cancer Res.. 91. 973-978 (2000)

[Publications] Kamiya,H., et al.: "2-Hydroxyadenine in DNA is a very poor substrate of the Escherichia coli MutY protein."J.Radiat.Res.. 41. 349-354 (2000)

[Publications] Fujikawa,K., et al.: "Human MTH1 protein hydrolyzes the oxidized ribonucleotide, 2-hydroxy-ATP."Nucleic Acids Res.. 29. 449-454 (2001)

[Publications] Kasai,H., et al.: "Life style and urinary 8-hydroxydeoxyguanosine, a marker of oxidative DNA damage : Effects of exercise, working conditions, meat intake, body mass index, and smoking."Jpn.J.Cancer Res.. 92. 9-15 (2001)

[Publications] Kasai,H., et al.: "Free Radicals in Chemistry, Biology and Medicine. (eds.Yoshikawa,T,et al)"Analysis of 8-hydroxydeoxyguanosine in cellular DNA and urine as a marker or oxidative stress.. 8 (2000)

葛西宏産業医科大学, 産業生態科学研究所, 教授 (40152615)