Metabolic activation of the environmental contaminant 3-nitrobenzanthrone by human acetyltransferases and sulfotransferase.
3-Nitrobenzanthrone (3-NBA) an extremely potent mutagen and suspected human carcinogen identified in diesel exhaust and in airborne particulate matter was shown to form multiple DNA adducts in vitro and in vivo in rats. In order to investigate whether human N,O-acetyltransferases (NATs) and sulfotransferases (SULTs) contribute to the metabolic activation of 3-NBA we used a panel of newly constructed Chinese hamster lung fibroblast V79MZ derived cell lines expressing human NAT1, human NAT2 or human SULT1A1, as well as TA1538-derived Salmonella typhimurium strains expressing human NAT1 (DJ400) or human NAT2 (DJ460) and determined DNA binding and mutagenicity. The formation of 3-NBA-derived DNA adducts was analysed by (32)P-postlabelling after exposing V79 cells to 0.01 micro M 3-NBA or 0.1 micro M N-acetyl-N-hydroxy-3-aminobenzanthrone (N-Ac-N-OH-ABA), a potential metabolite of 3-NBA. Similarly up to four major and two minor adducts were detectable for both compounds, the major ones being identical to those detected previously in DNA from rats treated with 3-NBA. Comparison of DNA binding between different V79MZ derived cells revealed that human NAT2 and, to a lesser extent, human NAT1 and human SULT1A1, contribute to the genotoxic potential of 3-NBA and N-Ac-N-OH-ABA to form DNA adducts. However, the extent of DNA binding by 3-NBA was higher in almost all V79 cells at a 10-fold lower concentration than by N-Ac-N-OH-ABA, suggesting that N-Ac-N-OH-ABA is not a major intermediate in the formation of 3-NBA-derived adducts. 3-NBA showed a 3.8-fold and 16.8-fold higher mutagenic activity in Salmonella strains expressing human NAT1 and human NAT2, respectively, than in the acetyltransferase-deficient strain, whereas N-Ac-N-OH-ABA was only clearly (but weakly) mutagenic in Salmonella DJ460 expressing human NAT2. This finding suggests that N-Ac-N-OH-ABA is not a major reactive metabolite responsible for the high mutagenic potency of 3-NBA in Salmonella. Collectively our results indicate that O-acetylation and O-sulfonation by human NATs and SULTs may contribute significantly to the high mutagenic and genotoxic potential of 3-NBA. Moreover, the yet-unidentified four major 3-NBA-derived adducts may be DNA adducts without an N-acetyl group.
Arlt
Volker M
VM
Institute of Cancer Research, Section of Molecular Carcinogenesis, Cotswold Road, Sutton, Surrey SM2 5NG, UK. v.arlt@icr.ac.uk
Glatt
Hansruedi
H
Muckel
Eva
E
Pabel
Ulrike
U
Sorg
Bernd L
BL
Schmeiser
Heinz H
HH
Phillips
David H
DH
D000393:Air Pollutants / Q000493:pharmacokinetics*; D000818:Animals; D001191:Arylamine N-Acetyltransferase / Q000378:metabolism*; D015239:Arylsulfotransferase; D001551:Benz(a)Anthracenes / Q000493:pharmacokinetics* / Q000494:pharmacology; D001711:Biotransformation; D002274:Carcinogens, Environmental / Q000493:pharmacokinetics*; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA / Q000187:drug effects; D018736:DNA Adducts; D004269:DNA, Bacterial / Q000187:drug effects; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism*; D015394:Molecular Structure; D009152:Mutagenicity Tests; D011355:Prodrugs / Q000493:pharmacokinetics*; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012486:Salmonella typhimurium / Q000187:drug effects; D013329:Structure-Activity Relationship; D015238:Sulfotransferases / Q000378:metabolism*; D014162:Transfection
D000393:Air Pollutants; D001551:Benz(a)Anthracenes; D002274:Carcinogens, Environmental; D018736:DNA Adducts; D004269:DNA, Bacterial; D007527:Isoenzymes; C469558:N-acetyl-N-hydroxy-3-aminobenzanthrone; D011355:Prodrugs; D011993:Recombinant Fusion Proteins; D004247:DNA; D001191:Arylamine N-Acetyltransferase; C089384:N-acetyltransferase 1; C478900:NAT2 protein, human; D015238:Sulfotransferases; D015239:Arylsulfotransferase; C117544:SULT1A1 protein, human; C117220:3-nitrobenzanthrone