The impact of NAT2 acetylator genotype on mutagenesis and DNA adducts from 2-amino-9H-pyrido[2,3-b]indole.
2-amino-9H-pyrido[2,3-b]indole (AalphaC) is a carcinogenic heterocyclic aromatic amine (HAA) that is produced in high quantities in tobacco smoke and that also forms in charred meats. The bioactivation of AalphaC occurs by cytochrome P450-mediated (P450 1A2) N-oxidation of the exocyclic amine group, to form 2-hydroxyamino-9H-pyrido[2,3-b]indole (HONH-AalphaC). The HONH-AalphaC metabolite can then undergo further activation by phase II enzymes to form the penultimate ester species, which bind to DNA. Some epidemiological studies suggest a role for NAT2 genetic polymorphisms in human susceptibilities to various cancers from tobacco smoke and from consumption of well-done meats, where the exposures to AalphaC can be substantial. In this investigation, we have measured the genotoxicity of AalphaC in nucleotide excision repair-deficient Chinese hamster ovary (CHO) cells stably transfected with human P450 1A2 and either the NAT2*4 (rapid, wild-type acetylator) or the NAT2*5B (the most common slow acetylator) allele, to determine the role of NAT2 phenotype in the biological effects of AalphaC. Mutations at the hypoxanthine phosphoribosyl transferase (hprt) locus were induced in a dose-dependent manner by AalphaC and were found to be highest in cells transfected with P450 1A2 and NAT2*4, followed by cells transfected with P450 1A2 and NAT2*5B. The level of formation of the deoxyguanosine (dG) adduct N-(deoxyguanosin-8-yl)-2-amino-9H-pyrido[2,3-b]indole (dG-C8-AalphaC) paralleled the mutagenic potency in these cell lines. However, AalphaC did not form DNA adducts or induce mutations in untransfected CHO cells or in cells only expressing P450 1A2. These findings clearly demonstrate that NAT2 genetic polymorphism plays a major role in the genotoxic potency of AalphaC.
Turesky
Robert J
RJ
Division of Environmental Health Sciences, Wadsworth Center, New York State Department of Health, Albany, New York 12201, USA. Rturesky@wadsworth.org
Bendaly
Jean
J
Yasa
Isil
I
Doll
Mark A
MA
Hein
David W
DW
D000818:Animals; D001191:Arylamine N-Acetyltransferase / Q000235:genetics / Q000378:metabolism*; D016466:CHO Cells; D002243:Carbolines / Q000737:chemistry* / Q000494:pharmacology / Q000633:toxicity; D002460:Cell Line; D002853:Chromatography, Liquid; D006224:Cricetinae; D003412:Cricetulus; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D018736:DNA Adducts / Q000737:chemistry* / Q000378:metabolism / Q000633:toxicity; D005838:Genotype; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics / Q000378:metabolism; D016296:Mutagenesis; D010641:Phenotype; D011110:Polymorphism, Genetic; D021241:Spectrometry, Mass, Electrospray Ionization; D014162:Transfection
D002243:Carbolines; D018736:DNA Adducts; D003577:Cytochrome P-450 Enzyme System; D001191:Arylamine N-Acetyltransferase; C478900:NAT2 protein, human; D007041:Hypoxanthine Phosphoribosyltransferase; C017822:2-amino-9H-pyrido(2,3-b)indole