Stereoselectivity of activation of 7,12-dimethylbenz[a]anthracene- 3,4-dihydrodiol to the anti-diol epoxide metabolite in a human mammary carcinoma MCF-7 cell-mediated V79 cell mutation assay.
7,12-Dimethylbenz[a]anthracene (DMBA), one of the most carcinogenic polycyclic aromatic hydrocarbons in rodent bioassays, is metabolically activated in many tissues to "bay-region" DMBA-3,4-diol-1,2-epoxides (DMBADE). Unlike benzo[a]pyrene, for which the high biological activity of the (7R,8S)-diol-(9S,10R)-epoxide has been established, the low chemical stability of anti-DMBADE has made it impossible to evaluate the role of specific stereoisomers in the biological activity of DMBA. In order to characterize the role of formation of DMBADE diastereomers in the induction of mutations, postlabeling assays using [35S]phosphorothioate with adduct separation by HPLC and immobilized boronate chromatography analyses were developed to allow separation and quantitation of DNA adducts formed from each stereoisomer of DMBADE. In DMBA-treated hamster embryo cell cultures, large quantities of three major adducts (anti-DMBADE-deoxyguanosine, anti-DMBADE-deoxyadenosine, and syn-DMBADE-deoxyadenosine) along with five minor adducts were completely resolved and quantitated. The DNA isolated from a human mammary carcinoma MCF-7 cell-mediated V79 cell mutation assay treated with increasing doses of racemic DMBA-3,4-dihydrodiol contained large amounts of two anti-DMBADE-DNA adducts. The anti-DMBADE adducts accounted for more than 90% of the total adducts at all doses. The number of 6-thioguanine-resistant mutants was proportional to the amount of anti-DMBADE-DNA adducts.(ABSTRACT TRUNCATED AT 250 WORDS)
Lau
H H
HH
Department of Medicinal Chemistry and Pharmacognosy, Purdue University, West Lafayette, Indiana 47907, USA.
Coffing
S L
SL
Lee
H
H
Harvey
R G
RG
Baird
W M
WM
D015127:9,10-Dimethyl-1,2-benzanthracene / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity; D000818:Animals; D001551:Benz(a)Anthracenes / Q000378:metabolism* / Q000633:toxicity; D001711:Biotransformation; D001943:Breast Neoplasms / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D006224:Cricetinae; D018736:DNA Adducts / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism; D004273:DNA, Neoplasm / Q000378:metabolism*; D005260:Female; D005347:Fibroblasts / Q000187:drug effects; D006801:Humans; D008168:Lung / Q000166:cytology; D016296:Mutagenesis / Q000187:drug effects*; D009152:Mutagenicity Tests; D009153:Mutagens / Q000737:chemistry / Q000378:metabolism* / Q000633:toxicity; D013237:Stereoisomerism; D014407:Tumor Cells, Cultured
C059444:7,12-dimethylbenz(a)anthracene-DNA adduct; D001551:Benz(a)Anthracenes; D018736:DNA Adducts; D004273:DNA, Neoplasm; D009153:Mutagens; C057927:dibenz(a,j)anthracene-3,4-diol-1,2-epoxide; D015127:9,10-Dimethyl-1,2-benzanthracene; C024024:7,12-dimethylbenz(a)anthracene-3,4-dihydrodiol