title: Oltipraz-mediated changes in aflatoxin B(1) biotransformation in rat liver: implications for human chemointervention.
abstract: Oltipraz (OPZ) is currently being considered for human use to protect against aflatoxin B1 (AFB)-induced hepatocarcinogenesis based on its proven protective effect in rats. The effectiveness of this treatment presumes that orthologous cytochrome P450 and glutathione S-transferase (GST) isozymes metabolize AFB in humans as they do in rats. In this study, alterations in the expression of multiple forms of cytochrome P450 and GST were evaluated after treatment with OPZ, as well as other known P450 inducers, including 3-methylcholanthrene, pregnenolone-16alpha-carbonitrile, and ciprofibrate. Evidence is presented that the male-specific rat CYP 3A2, an orthologue of human CYP 3A4, may be primarily responsible for AFB activation in rat liver at both high and low AFB substrate concentrations. The CYP 1A2 enzyme does not appear to play a role in AFB activation in rat liver at any substrate concentration, whereas the major human P450 enzyme capable of activating AFB at a low substrate concentration has been identified as CYP 1A2. Surprisingly, we found that the CYP 1A2 steady-state mRNA level and the CYP 1A2-associated methoxyresorufin-O-demethylase activity were induced approximately 3- and 2-fold, respectively, by OPZ in rat liver. However, because CYP 1A2 does not appear to participate in AFB activation, induction of CYP 1A2 may be insignificant for AFB-induced hepatocarcinogenesis in rat models. In the rat, a heterodimeric alpha class GST enzyme containing the Yc2 subunit is the only polypeptide characterized to date in this species with high catalytic activity for the conjugation of activated AFB with glutathione. The GST Yc2 steady-state mRNA level was induced 5-fold by OPZ treatment. This induction was mirrored by significant increases in both the corresponding protein level and AFB-8,9-epoxide-conjugating enzyme activity, which may contribute significantly to protection against AFB-induced carcinogenesis in the rat. Investigations from this and other laboratories have not revealed any evidence for a Yc2-like GST isozyme with high AFB-8,9-epoxide-conjugating activity in human liver. We have also been unable to demonstrate that the two major human alpha class GST isozymes, A1-1 and A2-2, purified from bacteria expressing the corresponding cDNAs, exhibit any significant AFB-8,9-epoxide-conjugating activity. Our results suggest that humans may not be protected to the same extent as rats against AFB-induced hepatocarcinogenesis by treatment with OPZ and that further investigations are needed to establish the usefulness of OPZ for protection against human exposure to AFB.
authors.affiliation: University of Washington, Department of Environmental Health, Seattle, 98195, USA.
authors.forename: T M
authors.initials: TM
authors.lastname: Buetler
authors.forename: T K
authors.initials: TK
authors.lastname: Bammler
authors.forename: J D
authors.initials: JD
authors.lastname: Hayes
authors.forename: D L
authors.initials: DL
authors.lastname: Eaton
mesh_terms: D016604:Aflatoxin B1 / Q000031:analogs & derivatives / Q000378:metabolism / Q000493:pharmacokinetics* / Q000633:toxicity; D000348:Aflatoxins / Q000378:metabolism; D000818:Animals; D016588:Anticarcinogenic Agents / Q000494:pharmacology*; D001483:Base Sequence; D001711:Biotransformation / Q000187:drug effects; D002273:Carcinogens / Q000493:pharmacokinetics*; D002995:Clofibric Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D019388:Cytochrome P-450 CYP1A2; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D004790:Enzyme Induction / Q000187:drug effects; D058607:Fibric Acids; D005982:Glutathione Transferase / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism*; D008099:Liver / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D008748:Methylcholanthrene / Q000494:pharmacology; D051379:Mice; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000378:metabolism*; D011285:Pregnenolone Carbonitrile / Q000494:pharmacology; D011719:Pyrazines / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D013045:Species Specificity; D013250:Steroid Hydroxylases / Q000378:metabolism*; D013871:Thiones; D013876:Thiophenes
chemical_list: D000348:Aflatoxins; D016588:Anticarcinogenic Agents; D002273:Carcinogens; D058607:Fibric Acids; D007527:Isoenzymes; D011719:Pyrazines; D011993:Recombinant Fusion Proteins; D013871:Thiones; D013876:Thiophenes; D011285:Pregnenolone Carbonitrile; C006426:aflatoxin B1-2,3-oxide; C033759:aflatoxin Q1; D002995:Clofibric Acid; D008748:Methylcholanthrene; C026209:oltipraz; D003577:Cytochrome P-450 Enzyme System; D016604:Aflatoxin B1; D010088:Oxidoreductases; D013250:Steroid Hydroxylases; D019388:Cytochrome P-450 CYP1A2; C022657:steroid hormone 6-beta-hydroxylase; D005982:Glutathione Transferase; C019304:ciprofibrate