Environmental pollutant and potent mutagen 3-nitrobenzanthrone forms DNA adducts after reduction by NAD(P)H:quinone oxidoreductase and conjugation by acetyltransferases and sulfotransferases in human hepatic cytosols.
3-Nitrobenzanthrone (3-nitro-7H-benz[de]anthracen-7-one, 3-NBA) is a potent mutagen and suspected human carcinogen identified in diesel exhaust and air pollution. We compared the ability of human hepatic cytosolic samples to catalyze DNA adduct formation by 3-NBA. Using the (32)P-postlabeling method, we found that 12/12 hepatic cytosols activated 3-NBA to form multiple DNA adducts similar to those formed in vivo in rodents. By comparing 3-NBA-DNA adduct formation in the presence of cofactors of NAD(P)H:quinone oxidoreductase (NQO1) and xanthine oxidase, most of the reductive activation of 3-NBA in human hepatic cytosols was attributed to NQO1. Inhibition of adduct formation by dicoumarol, an NQO1 inhibitor, supported this finding and was confirmed with human recombinant NQO1. When cofactors of N,O-acetyltransferases (NAT) and sulfotransferases (SULT) were added to cytosolic samples, 3-NBA-DNA adduct formation increased 10- to 35-fold. Using human recombinant NQO1 and NATs or SULTs, we found that mainly NAT2, followed by SULT1A2, NAT1, and, to a lesser extent, SULT1A1 activate 3-NBA. We also evaluated the role of hepatic NADPH:cytochrome P450 oxidoreductase (POR) in the activation of 3-NBA in vivo by treating hepatic POR-null mice and wild-type littermates i.p. with 0.2 or 2 mg/kg body weight of 3-NBA. No difference in DNA binding was found in any tissue examined (liver, lung, kidney, bladder, and colon) between null and wild-type mice, indicating that 3-NBA is predominantly activated by cytosolic nitroreductases rather than microsomal POR. Collectively, these results show the role of human hepatic NQO1 to reduce 3-NBA to species being further activated by NATs and SULTs.
Arlt
Volker M
VM
Section of Molecular Carcinogenesis, Institute of Cancer Research, Sutton, Surrey, United Kingdom. volker.arlt@icr.ac.uk
Stiborova
Marie
M
Henderson
Colin J
CJ
Osborne
Martin R
MR
Bieler
Christian A
CA
Frei
Eva
E
Martinek
Vaclav
V
Sopko
Bruno
B
Wolf
C Roland
CR
Schmeiser
Heinz H
HH
Phillips
David H
DH
D000123:Acetyltransferases / Q000378:metabolism*; D000818:Animals; D001551:Benz(a)Anthracenes / Q000378:metabolism* / Q000493:pharmacokinetics; D001711:Biotransformation; D003600:Cytosol / Q000201:enzymology / Q000378:metabolism; D018736:DNA Adducts / Q000096:biosynthesis*; D004785:Environmental Pollutants / Q000378:metabolism / Q000493:pharmacokinetics; D006801:Humans; D007527:Isoenzymes; D008099:Liver / Q000201:enzymology* / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008862:Microsomes, Liver / Q000201:enzymology / Q000378:metabolism; D008958:Models, Molecular; D009153:Mutagens / Q000378:metabolism* / Q000493:pharmacokinetics; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000378:metabolism*; D010084:Oxidation-Reduction; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D015238:Sulfotransferases / Q000378:metabolism*; D014969:Xanthine Oxidase / Q000378:metabolism
D001551:Benz(a)Anthracenes; D018736:DNA Adducts; D004785:Environmental Pollutants; D007527:Isoenzymes; D009153:Mutagens; D011994:Recombinant Proteins; D014969:Xanthine Oxidase; D016660:NAD(P)H Dehydrogenase (Quinone); C447479:NQO1 protein, human; D000123:Acetyltransferases; D015238:Sulfotransferases; C117220:3-nitrobenzanthrone