Effects of chronic dietary ethanol on in vivo and in vitro metabolism of methylazoxymethanol and on methylazoxymethanol-induced DNA methylation in rat colon and liver.
We examined the metabolism of 14C-labeled methylazoxymethanol (MAM) in male F344 rats pair-fed for 21.0 days either a liquid control diet, an isocaloric liquid diet containing 6.6% ethanol by volume (continuous ethanol diet), or the ethanol diet for 20.5 days followed by the control diet for 0.5 day (interrupted ethanol diet). Compared to rats fed the control liquid diet, metabolism of [1,2-14C]MAM acetate to exhaled 14CO2 was inhibited by 25 to 42% in rats fed the continuous ethanol diet, but was initially stimulated by 90% in rats given the interrupted ethanol diet. MAM-induced DNA methylation, as reflected in 7-methylguanine and O6-methylguanine content 24 h after carcinogen administration, was inhibited in the colon mucosae of rats fed the interrupted ethanol diet by 52 to 54%, and an even greater inhibition (71 to 86%) of DNA methylation occurred in the colon mucosae of rats fed the continuous ethanol diet. Liver DNA methylation was significantly inhibited (by 32 to 42%) only in those rats fed the continuous ethanol diet. Liver microsomes isolated from rats fed the 3 diets metabolized MAM to formic acid and methanol in vitro, but liver microsomes from rats fed the continuous ethanol diet were 12 to 15 times more active than liver microsomes from rats fed the control diet. Liver microsomes isolated from rats fed the interrupted ethanol diet were only 3 to 5 times more active in MAM metabolism than liver microsomes from rats fed the control diet, indicating very rapid turnover of the ethanol-induced enzyme(s) catalyzing the oxidation of MAM. Although chronic ethanol feeding enhanced the activity of liver microsomes for MAM metabolism, ethanol was found to inhibit the reaction competitively. Hepatocytes isolated from rats fed the continuous ethanol diet were considerably more sensitive to MAM-induced unscheduled DNA synthesis than hepatocytes isolated from rats given the control liquid diet, indicating that the stimulation of MAM metabolism by dietary ethanol results in increased DNA damage, observable in an in vitro system. Thus, the increased metabolic activation of MAM, due to enzyme induction by ethanol which is observed in vitro, is not reflected in increased liver DNA methylation in vivo.(ABSTRACT TRUNCATED AT 400 WORDS)
Fiala
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Naylor Dana Institute for Disease Prevention, Valhalla, New York 10595.
Sohn
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Hamilton
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D000437:Alcoholism / Q000378:metabolism*; D000818:Animals; D001391:Azo Compounds / Q000493:pharmacokinetics*; D002250:Carbon Radioisotopes; D002851:Chromatography, High Pressure Liquid; D003106:Colon / Q000187:drug effects / Q000378:metabolism*; D004247:DNA / Q000187:drug effects / Q000378:metabolism*; D004032:Diet; D000431:Ethanol / Q000494:pharmacology*; D007700:Kinetics; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D008745:Methylation; D008746:Methylazoxymethanol Acetate / Q000031:analogs & derivatives / Q000378:metabolism / Q000493:pharmacokinetics*; D051381:Rats; D011916:Rats, Inbred F344
D001391:Azo Compounds; D002250:Carbon Radioisotopes; D000431:Ethanol; D008746:Methylazoxymethanol Acetate; D004247:DNA; C030667:methylazoxymethanol