REGULAR ARTICLES
Genotoxic Activation of the Environmental Pollutant 3-Nitrobenzanthrone by Human Cytochrome P450 Enzymes Expressed in Salmonella typhimurium umu Tester Strains
2007 年 29 巻 4 号 p. 146-152
詳細
2007 年 29 巻 4 号 p. 146-152
3-Nitrobenzanthrone (3-NBA) is a mutagenic and carcinogenic compound identified in diesel exhaust, airborne particulate matter, soil, and water. To address whether 3-NBA shows genotoxic effects through any metabolic activation pathway, we determined the genotoxicity of 3-NBA using the parental strain Salmonella enterica serovar Typhimurium (S. typhimurium) TA1535/pSK1002, nitroreductase (NR)-deficient strain NM1000, the O-acetyltransferase (O-AT)-deficient strain NM2000, bacterial O-AT-overexpressing strain NM2009, and bacterial NR- and O-AT-overexpressing strain NM3009 established in our laboratory. In order to further clarify the role of human cytochrome P450 (P450 or CYP) enzyme in the bioactivation of 3-NBA to genotoxic metabolites, umu tester strains S. typhimurium OY1022/1A1, OY1022/1A2, OY1022/1B1, and OY1022/3A4, which express four respective human cytochrome P450 enzymes, NADPH-cytochrome P450 reductases (NPR) and O-AT were established by introducing two plasmids into S. typhimurium TA1535NR/1,8-DNP (nitroreductase-deficient and resistant to 1,8-dinitropyrene, probably due to functional loss of O-AT), one carrying both P450 and reductase cDNA in a biocistronic construct under control of an IPTG-inducible double tac promoter and the other, pOA102, carrying O-AT and umuC'-'lacZ fusion genes. Induction of umuC gene expression could be monitored by measuring the cellular β-galactosidase activity produced by the umuC'-'lacZ fusion gene. Although the induction of umuC by 3-NBA was weak in TA1535/pSK1002, NM1000, and NM2000 strains, the induction was considerably potent in NM2009 and NM3009 strains. 3-NBA was also found to induce umuC gene expression in OY1022/1A1 and OY1022/3A4 strains and, to a lesser extent, OY1022/1A2 and OY1022/1B1 strains, at a much higher rate than the parental OY1022/pCW strain. Collectively, these results suggest nitroreduction and O-acetylation by bacterial NR and O-AT, respectively, and nitroreduction by human CYP1A1, 3A4, 1A2, and 1B1 and NPR could contribute to the genotoxic activation of 3-NBA to produce reactive metabolites.
