Role of nitric oxide in the genotoxic response to chronic microcystin-LR exposure in human-hamster hybrid cells

Microcystin-LR (MC-LR) is the most abundant and toxic microcystin congener and has been classified as a potential human carcinogen (Group 2B) by the International Agency for Research on Cancer. However, the mechanisms underlying the genotoxic effects of MC-LR during chronic exposure are still poorly understood. In the present study, human-hamster hybrid (A(L)) cells were exposed to MC-LR for varying lengths of time to investigate the role of nitrogen radicals in MC-LR-induced genotoxicity. The mutagenic potential at the CD59 locus was more than 2-fold higher (p < 0.01) in A(L) cells exposed to a cytotoxic concentration (1 mu mol/L) of MC-LR for 30 days than in untreated control cells, which was consistent with the formation of micronucleus. MC-LR caused a dose-dependent increase in nitric oxide (NO) production in treated cells. Moreover, this was blocked by concurrent treatment with the NO synthase inhibitor N-G-methyl-L-arginine (L-NMMA), which suppressed MC-LRinduced mutations as well. The survival of mitochondrial DNA-depleted (rho(0)) A(L) cells was markedly decreased by MC-LR treatment compared to that in A(L) cells, while the CD59 mutant fraction was unaltered. These results provided clear evidence that the genotoxicity associated with chronic MC-LR exposure in mammalian cells was mediated by NO and might be considered as a basis for the development of therapeutics that prevent carcinogenesis. (C) 2015 The Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences.