Mitochondrial DNA adducts in the lung and liver of F344 rats chronically treated with 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone and (S)-4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol

摘要

Two recent studies conducted in our laboratory have demonstrated formation and accumulation of pyridyloxobutyl (POB) and pyridylhydroxybutyl (PHB) adducts in lung and liver total DNA of F344 rats chronically treated with the tobacco-specific carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and (R)- and (S)-enantiomers of its metabolite, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL). In this study, we measured POB and PHB adducts in lung and liver mitochondrial DNA (mtDNA), as previous studies suggest a potentially important role of mtDNA in carcinogenesis. Rats were sacrificed after 1, 2, 5, 10, 16, and 20 weeks of treatment with 10 ppm of NNK or (S)-NNAL in drinking water, and mtDNA and nuclear DNA (nDNA) adduct levels in lung and liver were determined by LC-ESI-MS/MS-SRM. The mean levels of individual POB adducts in mtDNA at all time points were slightly higher than those in nDNA for both NNK and (S)-NNAL-treated rats in the lung (P < 0.001 for both treatments), but not in the liver (P > 0.05). Lung mtDNA of both NNK- and (S)-NNAL-treated rats contained higher concentrations of the sum of three POB adducts (P < 0.001 for both treatments) than nDNA, while the levels of mtDNA and nDNA total POB adducts in the liver were not significantly different in either NNK- or (S)-NNAL-treated rats. Analysis of PHB adducts in mtDNA and nDNA produced results similar to those obtained for POB adducts. The steady accumulation of the lung and liver mtDNA adducts over the course of the study indicates inefficient repair of these adducts in mtDNA. This is the first study to examine the formation of NNK- and (S)-NNAL-derived adducts in rat mtDNA. The results support the hypothesis that preferential binding of tobacco carcinogens to mtDNA of the lung might be functionally important in the development of smoking-induced lung cancer.