title: Competitive inhibition of carcinogen-activating CYP1A1 and CYP1B1 enzymes by a standardized complex mixture of PAH extracted from coal tar.
abstract: A complex mixture of polycyclic aromatic hydrocarbons (PAH) extracted from coal tar, the Standard Reference Material (SRM) 1597, was recently shown to decrease the levels of DNA binding of the 2 strong carcinogens benzo[a]pyrene (BP) and dibenzo[a,l]pyrene (DBP) in the human mammary carcinoma-derived cell line MCF-7 (Mahadevan et al., Chem Res Toxicol 2005;18:224-231). The present study was designed to further elucidate the biochemical mechanisms involved in this inhibition process. We examined the effects of SRM 1597 on the metabolic activation of BP and DBP toward DNA-binding derivatives in Chinese hamster cells expressing either human cytochrome P450 (CYP) 1A1 or CYP1B1. SRM 1597 inhibited BP-DNA adduct formation through the entire exposure time in cells expressing human CYP1A1, while it significantly inhibited adduct formation only up to 48 hr when co-treated with DBP. Conversely, human CYP1B1-expressing cells were unable to catalyze PAH-DNA adduct formation on treatment with SRM 1597 alone, and on co-treatment with BP or DBP. The data obtained from biochemical experiments revealed that SRM 1597 competitively inhibited the activity of both human enzymes as analyzed by 7-ethoxyresorufin O-deethylation assays. While the Michaelis-Menten constant (K(M)) was <0.4 microM in the absence of SRM 1597, this value increased up to 1.12 (CYP1A1) or 4.45 microM (CYP1B1) in the presence of 0.1 microg/ml SRM 1597. Hence the inhibitory effects of the complex mixture on human CYP1B1 were much stronger when compared to human CYP1A1. Taken together, the decreases in PAH-DNA adduct formation on co-treatment with SRM 1597 revealed inhibitory effects on the CYP enzymes that convert carcinogenic PAH into DNA-binding metabolites. The implications for the tumorigenicity of complex environmental PAH mixtures are discussed.
authors.affiliation: Department of Environmental and Molecular Toxicology, Oregon State University, Corvallis, OR, USA.
authors.forename: Brinda
authors.initials: B
authors.lastname: Mahadevan
authors.forename: Charis P
authors.initials: CP
authors.lastname: Marston
authors.forename: Andreas
authors.initials: A
authors.lastname: Luch
authors.forename: Wan-Mohaiza
authors.initials: WM
authors.lastname: Dashwood
authors.forename: Eric
authors.initials: E
authors.lastname: Brooks
authors.forename: Cliff
authors.initials: C
authors.lastname: Pereira
authors.forename: Johannes
authors.initials: J
authors.lastname: Doehmer
authors.forename: William M
authors.initials: WM
authors.lastname: Baird
mesh_terms: D001189:Aryl Hydrocarbon Hydroxylases / Q000037:antagonists & inhibitors* / Q000187:drug effects; D001564:Benzo(a)pyrene / Q000037:antagonists & inhibitors / Q000633:toxicity; D001580:Benzopyrenes / Q000633:toxicity; D002273:Carcinogens / Q000037:antagonists & inhibitors* / Q000633:toxicity; D003033:Coal Tar / Q000737:chemistry*; D045424:Complex Mixtures / Q000302:isolation & purification / Q000494:pharmacology / Q000592:standards; D019363:Cytochrome P-450 CYP1A1 / Q000037:antagonists & inhibitors* / Q000187:drug effects; D065633:Cytochrome P-450 CYP1B1; D004247:DNA / Q000187:drug effects / Q000378:metabolism; D018736:DNA Adducts / Q000032:analysis; D006801:Humans; D011084:Polycyclic Aromatic Hydrocarbons / Q000302:isolation & purification / Q000494:pharmacology* / Q000592:standards; D014407:Tumor Cells, Cultured
chemical_list: D001580:Benzopyrenes; D002273:Carcinogens; D045424:Complex Mixtures; D018736:DNA Adducts; D011084:Polycyclic Aromatic Hydrocarbons; C518872:SRM 1597; D001564:Benzo(a)pyrene; D003033:Coal Tar; D004247:DNA; D001189:Aryl Hydrocarbon Hydroxylases; C585496:CYP1B1 protein, human; D019363:Cytochrome P-450 CYP1A1; D065633:Cytochrome P-450 CYP1B1; C041517:dibenzo(a,l)pyrene