Studies on DNA damages in vivo and in vitro by highly sensitive fluorometric analysis

通过高灵敏荧光分析研究体内和体外 DNA 损伤

• 批准号: 01044096
• 负责人: HAYATSU Hikoya
• 金额: $ 5.57万
• 依托单位: Okayama University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for international Scientific Research
• 财政年份: 1989
• 资助国家: 日本
• 起止时间: 1989 至 1991
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01044096/
• 关键词: fluorometry; DNA damage; N-nitrosamines; 8-methoxypsoralen; light; 近紫外光; 突然変異; 蛍光光度分析; DNAー本鎖切断; 光; 活性酸素; 遺伝子損傷阻害物質; フォスゲン; 螢光分析法; 細胞内DNA; ショウジョウバエ

DNA damages can be detected with high sensitivity by use of a fluorometric measurement. This method, originally developed by Morgan (the coinvestigator of this project), is based on the change in the fluorescence-intensity of ethidium bromide upon its intercalation into double-stranded DNA. Through our collaboration with Dr. Morgan, the following achievements were made.1. The fluorometric technique has been acquired by the Japanese workers, and the fluorometric measurement system is now established in Okayama University.2. Several new DNA-damaging agents were discovered. They include tetranitromethane, phosgene and N-nitrosamines with light.3. Cross-linking was detected in DNA that had been treated in vitro with 8-methoxypsoralen and UVA, but no cross-linking was found in the DNA of Drosophila lavae that had been treated with these agents in vivo. These observations indicate that DNA damages other than cross-linking must be responsible for the mutagenesis observed in the wing spots of the adult flies obtained from the lavae.

通过使用荧光测量，可以高灵敏度地检测 DNA 损伤。该方法最初由 Morgan（该项目的共同研究者）开发，基于溴化乙锭插入双链 DNA 时荧光强度的变化。通过与摩根博士的合作，我们取得了以下成果： 1．荧光技术已被日本工人掌握，目前冈山大学建立了荧光测量系统。 2．发现了几种新的 DNA 损伤剂。它们包括四硝基甲烷、光气和带光的N-亚硝胺。3．在体外用 8-甲氧基补骨脂素和 UVA 处理的 DNA 中检测到交联，但在用这些试剂体内处理的果蝇 DNA 中未发现交联。这些观察结果表明，在从熔岩中获得的成蝇翅膀斑点中观察到的诱变一定是交联以外的 DNA 损伤造成的。

项目成果

H. Hayatsu: "Dietary inhibitors of mutagenesis and carcinogenesis" Chemoprevention of Cancer, eds. by S. V. Bhide and G. B. Maru, Omega Scientific Publications, New Delhi. 40-50 (1992)

H. Hayatsu：“诱变和致癌的饮食抑制剂”，癌症化学预防，编辑。

T.Aoe,H.Hayatsu et al.: "DNA single-strand breaks induced in vitro by N-nitrosamines with irradiation of near ultraviolet light." Mutation Research,.

T.Aoe,H.Hayatsu 等人：“N-亚硝胺在近紫外光照射下在体外诱导 DNA 单链断裂。”

H. Hayatsu (ed.): CRC Press, FL. Mutagens in Food : Detection and Prevention, 1-286 (1991)

H. Hayatsu（编辑）：CRC Press，佛罗里达州。

H.Tsuchiyama,H.Hayatsu et al.: "Analysis of 2-amino-N^6-hydroxyadenine-induced mutagenesis in phage M13mp2" Mutation Research. 253. 47-54 (1991)

H.Tsuchiyama，H.Hayatsu 等人：“噬菌体 M13mp2 中 2-氨基-N^6-羟基腺嘌呤诱导的诱变分析”突变研究。

T.Negishi,H.Hayatsu et al.: "The genotoxicities in vivo of N-nitrosamines as assayed in the wing spot test and the DNA-repair test of Drosophila melanogaster" Mutation Research. 252. 119-128 (1991)

T.Negishi、H.Hayatsu 等人：“黑腹果蝇翅斑试验和 DNA 修复试验中测定的 N-亚硝胺体内基因毒性”突变研究。