Roles of DT diaphorase in the genotoxicity of nitroaromatic compounds in human and fish cell lines.
The genotoxicity of nitroaromatic compounds was examined in two cultured cell lines, namely, a human hepatoma cell line, HepG2, and a brown bullhead fibroblast cell line, BB. Furthermore, the role of the quinone-reducing enzyme DT diaphorase [NAD(P)H:(quinone acceptor) oxidoreductase] was examined with respect to its influence on the genotoxic effects of model nitroaromatic pollutants. The nitroreductive characteristics of these two cell lines were examined using an acetylated cytochrome c reduction assay for enzymatic nitroreductase activity. Subsequently, the influence of DT diaphorase on the genotoxicity of two model nitroaromatics, 4-nitroquinoline 1-oxide (4NQ) and nitrofurantoin (NF), revealed that DT diaphorase was the predominant 4NQ reductase in cytosols of both cell lines, but played a lesser role in NF reduction in both species. Despite these interspecific similarities, results revealed marked qualitative differences between the two species in terms of the influence of DT diaphorase on quinone-mediated genotoxicity. When pretreated with the DT diaphorase inhibitor dicoumarol, HepG2 cells exhibited an exacerbation of genotoxicity in the presence of 4NQ, indicating a protective influence of the enzyme. In contrast, 4NQ genotoxicity in BB cells was reduced in the presence of dicoumarol, indicating a deleterious effect of DT diaphorase activity. Conversely, dicoumarol pretreatment was moderately protective against NF-mediated genotoxicity in HepG2 cells but exacerbated NF toxicity in BB cells. This study illustrates the manner in which functionally analogous enzymes may have markedly distinct influences on xenobiotic toxicity in different cellular systems.
Hasspieler
B M
BM
Great Lakes Institute for Environmental Research, University of Windsor, Ontario, Canada.
Haffner
G D
GD
Adeli
K
K
D015112:4-Nitroquinoline-1-oxide / Q000633:toxicity*; D000818:Animals; D000892:Anti-Infective Agents, Urinary / Q000633:toxicity*; D002460:Cell Line; D004249:DNA Damage / Q000187:drug effects; D004277:DNA, Single-Stranded / Q000187:drug effects / Q000235:genetics; D001728:Dicumarol / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005399:Fishes; D006801:Humans; D009153:Mutagens / Q000633:toxicity*; D009154:Mutation / Q000187:drug effects / Q000235:genetics; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009582:Nitrofurantoin / Q000633:toxicity*; D011808:Quinone Reductases / Q000378:metabolism; D014407:Tumor Cells, Cultured
D000892:Anti-Infective Agents, Urinary; D004277:DNA, Single-Stranded; D004791:Enzyme Inhibitors; D009153:Mutagens; D015112:4-Nitroquinoline-1-oxide; D001728:Dicumarol; D009582:Nitrofurantoin; D016660:NAD(P)H Dehydrogenase (Quinone); D011808:Quinone Reductases