Mutagenic activation of carcinogenic N-nitrosopropylamines by rat liver: evidence for a cytochrome P-450 dependent reaction.
Mutagenic potential of carcinogenic N-nitrosopropylamines was examined by the Ames's liquid incubation assay, using rat liver 9000 g supernatant (S9) fraction for metabolic activation. N-Nitrosobis(2-hydroxypropyl)amine, N-nitroso(2-hydroxypropyl)-(2-oxopropyl)amine (HPOP), N-nitrosobis(2-oxopropyl)amine (BOP), N-nitrosobis(2-acetoxypropyl)amine, N-nitroso-2,6-dimethylmorpholine, N-nitrosomethyl-(2-hydroxypropyl)amine and N-nitrosomethyl(2-oxopropyl)amine all showed positive mutagenicity in strain TA100 in the presence of liver S9 while being negative in strain TA98. With the exception of HPOP and BOP, which were also mutagenic in TA100 without S9 metabolic activation, these N-nitrosopropylamines required the presence of microsomes as a source of enzymes as well as NADP+ as a cofactor for mutagenic activation. Treatment of rats with polychlorinated biphenyls or phenobarbital (PB) resulted in a marked increase in the ability of S9 to activate the seven N-nitrosamines tested whereas 3-methylcholanthrene (3-MC) induction was not effective. All the mutagenic activities were considerably decreased by preincubation in an atmosphere of either carbon monoxide or nitrogen gas or by adding cytochrome c to the S9 mixture. Metyrapone, a specific inhibitor of PB-inducible major cytochrome P-450, considerably inhibited mutagenicity, whereas 7,8-benzoflavone, a specific inhibitor of 3-MC-inducible major cytochrome P-448, was totally lacking this effect. These results demonstrate a correlation between rat liver S9 dependent mutagenicity of six N-nitrosopropylamines and their known carcinogenicity in rat in vivo experiments, and that the PB-inducible major cytochrome P-450 is involved in the mutagenic activation. BOP was also shown to be activated by extrahepatic (lung, kidney, pancreas) tissue S9, blood S9 and bovine serum albumin (BSA) to the extent of 50% of that activity obtained with liver S9. A possible mechanism of BSA-mediated activation of BOP is discussed.
D000818:Animals; D001711:Biotransformation; D002273:Carcinogens / Q000378:metabolism*; D003577:Cytochrome P-450 Enzyme System / Q000502:physiology*; D005954:Glucosephosphate Dehydrogenase / Q000494:pharmacology; D066298:In Vitro Techniques; D008297:Male; D008862:Microsomes, Liver / Q000378:metabolism*; D009153:Mutagens / Q000378:metabolism*; D009602:Nitrosamines / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D012486:Salmonella typhimurium / Q000187:drug effects
D002273:Carcinogens; D009153:Mutagens; D009602:Nitrosamines; C012457:diisopropanolnitrosamine; C013622:nitrosobis(2-oxopropyl)amine; D003577:Cytochrome P-450 Enzyme System; D005954:Glucosephosphate Dehydrogenase