Puck T T, Johnson R, Webb P, Yohrling G
Eleanor Roosevelt Institute, Denver, Colorado 80206, USA.
Somat Cell Mol Genet. 1998 Jan;24(1):1-11. doi: 10.1007/BF02677491.
The interaction of chemical mutagens with mammalian cells is much more complex than that of gamma-irradiation because of the different ways in which chemical agents react with cell and medium components. Nevertheless, the system previously described for analysis of mutagenesis by gamma-radiation appears applicable to chemical mutagenesis. The approach involves measurement of cell survival, use of caffeine to inhibit repair, analysis of mitotic index changes, and quantitation of microscopically visible structural changes in mitotic chromosomes. The behavior of a variety of chemical mutagens and nonmutagens in this system is described and compared with that of gamma-irradiation. The procedure is simple and the results reasonably quantitative though less so than those of gamma-irradiation. The procedure can be used for environmental monitoring, analysis of mutational events, and individual and epidemiological testing. Mutational events should be classified as primary or secondary depending on whether they represent initial genomic insult, or genomic changes resulting from primary mutation followed by structural changes due to metabolic actions. While caffeine has multiple effects on the mammalian genome, when used under the conditions specified here it appears to act principally as an inhibitor of mutation repair, and so affords a measure of the role of repair in the action of different mutagens on cells in the G2 phase of the life cycle.
由于化学诱变剂与细胞及培养基成分的反应方式不同，其与哺乳动物细胞的相互作用比γ射线照射更为复杂。然而，先前描述的用于分析γ射线诱变作用的系统似乎也适用于化学诱变作用。该方法包括测量细胞存活率、使用咖啡因抑制修复、分析有丝分裂指数变化以及对有丝分裂染色体微观可见的结构变化进行定量。描述了多种化学诱变剂和非诱变剂在该系统中的行为，并与γ射线照射的行为进行了比较。该方法简单，结果具有一定的定量性，尽管不如γ射线照射的结果精确。该方法可用于环境监测、突变事件分析以及个体和流行病学检测。突变事件应根据其是否代表初始基因组损伤，或是否为由初级突变导致的基因组变化以及随后因代谢作用引起的结构变化，分为原发性或继发性。虽然咖啡因对哺乳动物基因组有多种影响，但在此处规定的条件下使用时，它似乎主要作为突变修复的抑制剂起作用，因此可衡量修复在不同诱变剂对生命周期G2期细胞作用中的作用。
