A benzo[alpha]pyrene-7,8-dihydrodiol-9,10-epoxide is the major metabolite involved in the binding of benzo[alpha]pyrene to DNA in isolated viable rat hepatocytes.
Benzo[alpha]pyrene is metabolised by isolated viable hepatocytes from both untreated and 3-methylcholanthrene pretreated rats to reactive metabolites which covalently bind to DNA. The DNA from the hepatocytes was isolated, purified and enzymically hydrolysed to deoxyribonucleosides. The hydrocarbon-deoxyribonucleoside products after initial separation, on small columns of Sephadex LH-20, from unhydrolysed DNA, oligonucleotides and free bases, were resolved by high pressure liquid chromatography (HPLC). The qualitative nature of the adducts found in both control and pretreated cells was virtually identical; however pretreatment with 3-methylcholanthrene resulted in a quantitatively higher level of binding. The major hydrocarbon-deoxyribonucleoside adduct, found in hepatocytes co-chromatographed with that obtained following reaction of the diol-epoxide, (+/-) 7 alpha,8 beta-dihydroxy-9 beta,10 beta-epoxy-7,8,9,10-tetrahydrobenzo[alpha]pyrene with DNA. Small amounts of other adducts were also present including a more polar product which co-chromatographed with the major hydrocarbon-deoxyribonucleoside adduct formed following microsomal activation of 9-hydroxybenzo[alpha]-pyrene and subsequent binding to DNA. In contrast to the results with hepatocytes, when microsomes were used to metabolically activate benzo[alpha]-pyrene, the major DNA bound-product co-chromatographed with the more polar adduct formed upon further metabolism of 9-hydroxybenzo[alpha]pyrene. These results illustrate that great caution must be exercised in the extrapolation of results obtained from short-term mutagenesis test systems, utilising microsomes, to in vivo carcinogenicity studies.
D000818:Animals; D001580:Benzopyrenes / Q000378:metabolism* / Q000633:toxicity; D001665:Binding Sites; D001711:Biotransformation; D002273:Carcinogens; D004247:DNA / Q000378:metabolism*; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004852:Epoxy Compounds / Q000378:metabolism; D005260:Female; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D008748:Methylcholanthrene / Q000494:pharmacology; D009153:Mutagens; D051381:Rats
D001580:Benzopyrenes; D002273:Carcinogens; D004852:Epoxy Compounds; D009153:Mutagens; D008748:Methylcholanthrene; D004247:DNA