Repair of DNA adducts in asynchronous CHO cells and the role of repair in cell killing and mutation induction in synchronous cells treated with 7-bromomethylbenz[a]anthracene.
CHO cells of normal or UV-sensitive phenotypes were analyzed for their ability to remove DNA adducts produced by the carcinogen 7-BrMeBA. At a dose of 0.1 microM, which reduced the survival of the normal AA8 cells to approximately 90% and the mutant UV5 cells to approximately 20%, the frequency of adducts was 5-6 per 10(6) nucleotides for both cell types, and AA8 cells removed approximately 30% of the adducts in 8 h and approximately 55% in 24 h. In contrast, UV5 and mutants from four other genetic complementation groups had no significant removal. Binding of 7-BrMeBA did not vary through the cell cycle in synchronous cultures. At a dose of mutagen (0.07 microM) resulting in approximately 25% survival of asynchronous UV5, the survival of synchronous cultures rose about threefold from early G1 to early S phase and then decreased somewhat in late S/G2. At a dose (0.28 microM) producing similar survival of asynchronous cultures, AA8 cells differed qualitatively in that survival decreased progressively by 5- to 10-fold between early G1 and the early part of S, and rose steeply through late S/G2 to give a 10- to 20-fold increase. We conclude that DNA repair is the major determinant of variations in survival through the cycle in normal cells. The patterns observed are consistent with a mechanism of killing in AA8 cells in which adducts disrupt DNA replication, while in UV5 cells transcriptional blocks or other effects may govern lethality. Induced mutations at the aprt and hprt loci showed changes through the cycle in both AA8 and UV5 cells, and the patterns were not readily explainable by the action of repair.
D000818:Animals; D001551:Benz(a)Anthracenes / Q000633:toxicity*; D002453:Cell Cycle / Q000187:drug effects; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA / Q000378:metabolism*; D004260:DNA Repair; D005260:Female; D007700:Kinetics; D009154:Mutation; D010053:Ovary
D001551:Benz(a)Anthracenes; C001903:7-bromomethylbenzanthracene; D004247:DNA