Akiyama N, Murata S, Alexander D B, Yaoita H, Aoki Y, Noda M
Department of Molecular Oncology, Kyoto University Graduate School of Medicine, Japan.
J Epidemiol. 1999 Dec;9(6 Suppl):S72-7. doi: 10.2188/jea.9.6sup_72.
A retroviral vector carrying both positive (neo) and negative (herpes simplex virus thymidine kinase or HSV-tk) selection markers was constructed as a substrate for mutational assay in mammalian cells. Using a population of rat fibroblast cells carrying a single copy per cell of retroviral DNA randomly integrated in their chromosomes, we examined the cytotoxic and mutagenic activities of ultraviolet light (UV) at four wavelengths (254, 290, 300, and 320 nm). The action spectra for these activities are similar to some of the previously reported spectra for photochemical DNA modifications, erythema, cell killing, and mouse skin carcinogenesis, except at 290 and 320 nm. At 290 nm, no significant mutagenicity was observed. At 320 nm, both cytotoxic and mutagenic activities were 10 times higher than the values expected from the absorption spectrum for DNA and the action spectrum for bacterial inactivation and mutagenesis. Structural comparison of some of the HSV-tk mutants obtained after irradiation with 300 and 320 nm UV revealed partially different patterns of mutation specificity, suggesting the involvement of multiple molecular mechanisms in the genotoxicity associated with this range of UV.
构建了一种携带正向(新霉素抗性基因,neo)和负向(单纯疱疹病毒胸苷激酶,HSV - tk)选择标记的逆转录病毒载体,作为哺乳动物细胞突变检测的底物。利用每个细胞携带一个随机整合在其染色体中的逆转录病毒DNA单拷贝的大鼠成纤维细胞群体,我们检测了四种波长(254、290、300和320纳米)的紫外线(UV)的细胞毒性和诱变活性。这些活性的作用光谱与之前报道的一些光化学DNA修饰、红斑、细胞杀伤和小鼠皮肤致癌作用的光谱相似,但在290和320纳米处除外。在290纳米处,未观察到明显的诱变活性。在320纳米处,细胞毒性和诱变活性均比根据DNA吸收光谱以及细菌失活和诱变作用光谱所预期的值高10倍。对用300和320纳米紫外线照射后获得的一些HSV - tk突变体进行结构比较,发现突变特异性模式部分不同,这表明在与该范围紫外线相关的遗传毒性中涉及多种分子机制。