Regulation of the formation of the major diethylstilbestrol-DNA adduct and some evidence of its structure.
Diethylstilbestrol (DES) induces kidney tumors in hamsters. In previous studies, DES has been shown by 32P-post-labeling analysis to bind covalently to DNA in vivo and in vitro and DES-DNA adduct formation has been suggested to play a key role in DES-induced carcinogenicity. In this study, we have examined the influence of the dose of DES, age of animals and organ specificity on adduct formation in hamsters. In addition, we examined the characteristics of DES-DNA adduct formation in vitro and the structure of the major adduct. DES-DNA adducts were detected in liver and kidney of hamsters treated with at least 20 mg/kg DES. Adduct concentrations were higher at higher doses or in older compared to younger animals. The covalent binding of DES to DNA catalyzed by hamster liver microsomes required cumene hydroperoxide as cofactor, whereas with NADPH, adducts were barely detectable, presumably because the reactive metabolic intermediate DES quinone was reduced to DES. The major DES-DNA adduct formed in vitro was purified by semipreparative and analytical high pressure liquid chromatography. It is concluded that DES-DNA adducts are formed from DES quinone at very low rates in vitro and occur at low levels in vivo, even when hamsters receive very large doses of DES. The dependence of DES-DNA adduct concentrations in vitro on organic hydroperoxide cofactors required for cytochrome P450-mediated DES quinone formation indicates that stilbene-DNA adduction may occur only under conditions of oxidative stress.
Bhat
H K
HK
Department of Pharmacology and Toxicology, University of Texas Medical Branch, Galveston 77555-1031.
Han
X
X
Gladek
A
A
Liehr
J G
JG
D000818:Animals; D001555:Benzene Derivatives / Q000378:metabolism; D006224:Cricetinae; D004247:DNA / Q000187:drug effects* / Q000378:metabolism*; D018736:DNA Adducts / Q000096:biosynthesis* / Q000302:isolation & purification / Q000493:pharmacokinetics; D004054:Diethylstilbestrol / Q000031:analogs & derivatives* / Q000378:metabolism* / Q000494:pharmacology; D004355:Drug Stability; D005260:Female; D006207:Half-Life; D008297:Male; D008647:Mesocricetus; D008862:Microsomes, Liver / Q000378:metabolism; D009249:NADP / Q000378:metabolism; D010084:Oxidation-Reduction
D001555:Benzene Derivatives; D018736:DNA Adducts; D009249:NADP; C039998:diethylstilbestrol quinone; D004054:Diethylstilbestrol; D004247:DNA; C007164:cumene hydroperoxide