Mutagenicity of 7H-dibenzo[c,g]carbazole and its tissue specific derivatives in genetically engineered Chinese hamster V79 cell lines stably expressing cytochrome P450

Genetically engineered Chinese hamster V79 cell lines with stable expression of human cytochrome P4501A1 and 1A2 were used to characterize the particular form of P450 enzymes capable of activating 7H-dibenzo[c,g]carbazole (DBC) and its tissue- and organ-specific derivatives, N-methylDBC (N-MeDBC) and 5,9-dimethylDBC (diMeDBC). In addition, a V79 cell line with co-expression of CYP1A2 together with a phase H enzyme, N-acetyltransferase was utilized to study the role of an entire metabolic activation system in biotransformation of these carbazoles. The rise of 6-thioguanine resistant (6-TG') mutations was followed as a marker of biological activity of these agents. None of the carbazoles elevated significantly the frequency of mutations in the parental V79MZ cell line lacking any cytochrome P450 (CYP) activity or in the V79NH cells expressing N-acetyltransferase activity. A variable, however, increase of mutations was found in the cell lines expressing CYP activity. Both DBC, a potent liver and skin carcinogen, and N-MeDBC, a specific sarcomagen, increased significantly (P < 0.001) the frequency of 6-TG(r) mutations in V79MZh1A1 cells, expressing the human CYP2A1; in contrast, a strict hepatocarcinogen diMeDBC was devoid of any activity. All carbazoles elevated significantly the level of mutations in the V79MZh1A2 cell line expressing the human CYP1A2, N-MeDBC was most efficient. Co-expression of CYP1A2 together with NAT activity significantly reduced or totally eliminated the mutagenicity of all carbazoles. These data confirm that CYP1A1 is explicitly involved in the activation of sarcomagenic DBC derivatives, whereas CYP1A2 is included in biotransformation of all DBC derivatives. Reactive intermediates formed due to CYP1A2 activation are substrate for conjugation reactions mediated by N-acetyltransferase. (C) 2002 Elsevier Science B.V All rights reserved.