1997 年度実績報告書

酸化的DNA損傷の生成と修復

研究課題

研究課題/領域番号	08264237
研究機関	産業医科大学
研究代表者	葛西宏産業医科大学, 産業生態科学研究所, 教授 (40152615)
研究分担者	紙谷浩之産業医科大学, 産業生態科学研究所, 助手 (10204629) 平野雄産業医科大学, 産業生態科学研究所, 助手 (40258629)
キーワード	8-ヒドロキシグアニン / 2-ヒドロキシアデニン / 活性酸素 / グリオキサール
研究概要	酸化的DNA環傷の生成、変異誘発、修復について以下の点を明らかにした。 1.喫煙と肺の酸化ストレスの関係を明らかにするためにヒト肺中心部の組織からDNAを抽出しHPLC-ECD法により8-ヒドロキシグアニン(8-OH-Gua)を側滑走した。喫煙者は非喫煙者に比ベ8-OH-Gua量が有意に高かった(p=0.026)。また8-OH-Gua量とBrinkman indexとの間に正の相関が見られた(r=0.53,p=0.013)。 2.運動と活性酸素の関係を調べるためにラットに強制運動をさせた群、自発運動をさせた群、および運動をさせた群、および運動させなかった群について、肝臓、腎臓、心臓DNA中の8-OH-Gua量を測定した。その結果、強制運動ラット群では自発運動群に比ベ3臓器とも高い8-OH-Gua値を示した(p<0.05)。 3.内因性変異原グリオキサールの変異スペクトルを大腸菌および哺乳動物細胞を用いて調べた。その結果、グリオキサールは大腸菌、哺乳動物細胞中のいずれの場合にもG:C塩基対における塩基置換変異を誘発した。大腸菌では、G:C→A:T、G:C→T:A、G:C→C:Gの順に変異が誘発された。哺乳動物細胞ではG:C→T:A、G:C→C:G、G:C→A:Tの順に変異が誘発された。 4.シャトルベクターを用いて、2-ヒドロキシアデニン(2-OH-Ade)が哺乳動物細胞(COS-7)で誘発する変異の解析を行った。その結果、大腸菌と同様に、2-OH-Adeは、8-OH-Guaと同程度の変異誘発能(導入した2-OH-Adeの約1%が変異を誘発)を示した。誘発した変異はA→G及びAの欠失であった。 5.ヒト8-OH-Gua修復酵素遺伝子hOGGlのクローニングがなされたが、本実験ではこの遺伝子産物が実際に8-OH-Guaグリコシラーゼ活性を持つかどうかを調べた。GST-hOGGlタンパクの作用によりDNAから8-OH-Guaが遊離するかどうかをHPLC-ECD法により調べたところ、8-OH-Gua・Cおよび8-OH-Gua塩基対を含むDNAに対し活性が見られた。

研究成果
(19件)

すべてその他

すべて文献書誌 (19件)

[文献書誌] Kamiya,H., et al.: "Substitution and deletion mutations induced by 2-hydroxyadenine in Escherichia coli:Effects of sequence contexts in leading and lagging strands." Nucleic Acids Res.25. 304-310 (1997)
[文献書誌] Murata-Kamiya,N., et al.: "Types of mutations induced by glyoxal,a major oxidative DNA-damage product,in Salmonella typhimurium." Mutation Res.377. 13-16 (1997)
[文献書誌] Murata-Kamiya,N., et al.: "Mutational specificity of glyoxal,a product of DNA oxidation,in the lacI gene of wild-type Escherichia coli W3110." Mutation Res.377. 255-262 (1997)
[文献書誌] Murata-Kamiya,N., et al.: "Glyoxal,a major product of DNA oxidation,induces mutations at G:C sites on a shuttle vector plasmid replicated in mammalian cells." Nucleic Acid Res.25. 1897-1902 (1997)
[文献書誌] Nagashima,M., et al.: "Presence of human cellular protein(s) that specifically binds and cleaves 8-hydroxyguanine containing DNA." Mutation Res.383. 49-59 (1997)
[文献書誌] Takahashi,K., et al.: "Relationship between asbestos exposures and 8-hydroxydeoxyguanosine levels in leukocytic DNA of workers at a Chinese asbestos-material plant." Int.J.Occup.Environ.Health. 3. 111-119 (1997)
[文献書誌] Zhang,D., et al.: "Vitamin E inhibits apoptosis,DNA modfication,and cancer incidence induced by iron-mediated peroxidation in Wister rat kidney." Cancer Res.57. 2410-2414 (1997)
[文献書誌] Asami,S., et al.: "Cigarette smoking induces an increase in oxidative DNA damage,8-hydroxydeoxyguanosine,in a central site of the human lung." Carcinogenesis. 18. 1763-1766 (1997)
[文献書誌] Kamiya,H., et al.: "Mutations induced by 2-hydroxyadenine on a shuttle vector during leading and lagging strand syntheses in mammalian cells." Biochemistry. 36. 11125-11130 (1997)
[文献書誌] Nagashima,M., et al.: "Presence of human cellular protein(s) that specifically binds and cleaves 8-hydroxydeoxyguanine containing DNA." Mutation Res.383. 49-59 (1997)
[文献書誌] Hirano,T., et al.: "Inhibition of 8-hydroxyguanine repair in testis after administration of cadmium chloride to GSH-depleted rats." Toxicology and Applied Pharmacology. 147. 9-14 (1997)
[文献書誌] Shinmura,K., et al.: "8-hydroxyguanine (7,8-dihydro-8-oxoguanine)DNA glycosylase and AP lyase activities of hOGG1 protein and their substrate specificity." Mutation Res.385. 75-82 (1997)
[文献書誌] Suzuki,M., et al.: "Construction of mutants of Salmonella typhymurium deficient in 8-hydroxyguanine DNA glycosylase and their sensitivities to oxidative mutagens and nitro compounds." Mutation Res.393. 233-246 (1997)
[文献書誌] Kasai,H.: "Analysis of a form of oxidative DNA damage,8-hydroxy-2'-deoxyguanosine,as a marker of cellular oxidative stress during carcinogenesis." Mutation Res.387. 147-163 (1997)
[文献書誌] Tani,M., et al.: "Genomic structure and chromosomal localozation of the mouse Ogg1 gene that is involved in the repair of 8-hydroxyguanine in DNA damage." Mamm Genome. 9. 32-37 (1998)
[文献書誌] Kamath-Loeb,A.S., et al.: "Incorporation of the guanosine triphosphate analogs 8-oxo-dGTP and 8-NH_2-dGTP by reverse transcriptases and mammalian DNA polymerases." J.Biol.Chem.272. 5892-5898 (1997)
[文献書誌] Murata-Kamiya,N., et al.: "Comparision of oxidation products from DNA components by γ-irradiation and Fenton-type reactions." J.Radiat.Res.38. 121-131 (1997)
[文献書誌] Asami,S., et al.: "Efects of forced and spontaneous exercise on 8-hydroxydeoxyguanine levels in rat organs." Biochem.Biophys.Res.Commun.(in press). (1998)
[文献書誌] Kasai,H., et al.: "Food Factors for Cancer Prevention (In creased 8-hydroxyguanine repair activity in animal livers and human leukocytes induced by oxidative stress;........)" Springer-Verlag Tokyo, 5 (1997)

1997 年度 実績報告書

酸化的DNA損傷の生成と修復

研究代表者

葛西 宏 産業医科大学, 産業生態科学研究所, 教授 (40152615)

研究成果

[文献書誌] Kamiya,H., et al.: "Substitution and deletion mutations induced by 2-hydroxyadenine in Escherichia coli:Effects of sequence contexts in leading and lagging strands." Nucleic Acids Res.25. 304-310 (1997)

[文献書誌] Murata-Kamiya,N., et al.: "Types of mutations induced by glyoxal,a major oxidative DNA-damage product,in Salmonella typhimurium." Mutation Res.377. 13-16 (1997)

[文献書誌] Murata-Kamiya,N., et al.: "Mutational specificity of glyoxal,a product of DNA oxidation,in the lacI gene of wild-type Escherichia coli W3110." Mutation Res.377. 255-262 (1997)

[文献書誌] Murata-Kamiya,N., et al.: "Glyoxal,a major product of DNA oxidation,induces mutations at G:C sites on a shuttle vector plasmid replicated in mammalian cells." Nucleic Acid Res.25. 1897-1902 (1997)

[文献書誌] Nagashima,M., et al.: "Presence of human cellular protein(s) that specifically binds and cleaves 8-hydroxyguanine containing DNA." Mutation Res.383. 49-59 (1997)

[文献書誌] Takahashi,K., et al.: "Relationship between asbestos exposures and 8-hydroxydeoxyguanosine levels in leukocytic DNA of workers at a Chinese asbestos-material plant." Int.J.Occup.Environ.Health. 3. 111-119 (1997)

[文献書誌] Zhang,D., et al.: "Vitamin E inhibits apoptosis,DNA modfication,and cancer incidence induced by iron-mediated peroxidation in Wister rat kidney." Cancer Res.57. 2410-2414 (1997)

[文献書誌] Asami,S., et al.: "Cigarette smoking induces an increase in oxidative DNA damage,8-hydroxydeoxyguanosine,in a central site of the human lung." Carcinogenesis. 18. 1763-1766 (1997)

[文献書誌] Kamiya,H., et al.: "Mutations induced by 2-hydroxyadenine on a shuttle vector during leading and lagging strand syntheses in mammalian cells." Biochemistry. 36. 11125-11130 (1997)

[文献書誌] Nagashima,M., et al.: "Presence of human cellular protein(s) that specifically binds and cleaves 8-hydroxydeoxyguanine containing DNA." Mutation Res.383. 49-59 (1997)

[文献書誌] Hirano,T., et al.: "Inhibition of 8-hydroxyguanine repair in testis after administration of cadmium chloride to GSH-depleted rats." Toxicology and Applied Pharmacology. 147. 9-14 (1997)

[文献書誌] Shinmura,K., et al.: "8-hydroxyguanine (7,8-dihydro-8-oxoguanine)DNA glycosylase and AP lyase activities of hOGG1 protein and their substrate specificity." Mutation Res.385. 75-82 (1997)

[文献書誌] Suzuki,M., et al.: "Construction of mutants of Salmonella typhymurium deficient in 8-hydroxyguanine DNA glycosylase and their sensitivities to oxidative mutagens and nitro compounds." Mutation Res.393. 233-246 (1997)

[文献書誌] Kasai,H.: "Analysis of a form of oxidative DNA damage,8-hydroxy-2'-deoxyguanosine,as a marker of cellular oxidative stress during carcinogenesis." Mutation Res.387. 147-163 (1997)

[文献書誌] Tani,M., et al.: "Genomic structure and chromosomal localozation of the mouse Ogg1 gene that is involved in the repair of 8-hydroxyguanine in DNA damage." Mamm Genome. 9. 32-37 (1998)

[文献書誌] Kamath-Loeb,A.S., et al.: "Incorporation of the guanosine triphosphate analogs 8-oxo-dGTP and 8-NH_2-dGTP by reverse transcriptases and mammalian DNA polymerases." J.Biol.Chem.272. 5892-5898 (1997)

[文献書誌] Murata-Kamiya,N., et al.: "Comparision of oxidation products from DNA components by γ-irradiation and Fenton-type reactions." J.Radiat.Res.38. 121-131 (1997)

[文献書誌] Asami,S., et al.: "Efects of forced and spontaneous exercise on 8-hydroxydeoxyguanine levels in rat organs." Biochem.Biophys.Res.Commun.(in press). (1998)

[文献書誌] Kasai,H., et al.: "Food Factors for Cancer Prevention (In creased 8-hydroxyguanine repair activity in animal livers and human leukocytes induced by oxidative stress;........)" Springer-Verlag Tokyo, 5 (1997)

1997 年度実績報告書

葛西宏産業医科大学, 産業生態科学研究所, 教授 (40152615)