Autoxidation of extracellular hydroquinones is a causative event for the cytotoxicity of menadione and DMNQ in A549-S cells.
Cytotoxicity of 1,4-naphthoquinones has been attributed to intracellular reactive oxygen species (ROS) generation through one-electron-reductase-mediated redox cycling and to arylation of cellular nucleophiles. Here, however, we report that in a subclone of lung epithelial A549 cells (A549-S previously called A549-G4S (Watanabe, et al., Am. J. Physiol. 283 (2002) L726-736), the mechanism of ROS generation by menadione and by 2,3-dimethoxy-1,4-naphthoquinone (DMNQ), and therefore that of cytotoxicity, differs from the paradigm. Ninety percent of H(2)O(2) generation by both the quinones can be prevented by dicumarol, an inhibitor of NAD(P)H quinone oxidoreductase (NQO1), at the submicromolar level, regardless of the quinone concentrations. Exogenous SOD also inhibits H(2)O(2) production at low but not high concentrations of the quinones, especially DMNQ. Thus, at low quinone concentrations, superoxide-driven hydroquinone autoxidation accounts for more than half of H(2)O(2) generation by both quinones, whereas at high quinone concentrations, especially for DMNQ, comproportionation-driven hydroquinone autoxidation becomes the predominant mechanism. Hydroquinone autoxidation appears to occur predominantly in the extracellular environment than in the cytosol as extracellular catalase can dramatically attenuate quinone-induced cytotoxicity throughout the range of quinone concentrations, whereas complete inactivation of endogenous catalase or complete depletion of intracellular glutathione has only a marginal effect on their cytotoxicity. Finally, we show evidence that ROS production is a consequence of the compensatory defensive role of NQO1 against quinone arylation.
Watanabe
Nobuo
N
Department of Environmental Health Sciences, School of Public Health and Center for Free Radical Biology, University of Alabama at Birmingham, RPHB-534, 1530 3RD Avenue South, Birmingham, AL 35294-0022, USA.
Forman
Henry Jay
HJ
D019328:Buthionine Sulfoximine / Q000494:pharmacology; D002470:Cell Survival / Q000187:drug effects*; D002999:Clone Cells; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D006873:Hydroquinones / Q000378:metabolism*; D008168:Lung; D009285:Naphthoquinones / Q000633:toxicity*; D010084:Oxidation-Reduction; D017382:Reactive Oxygen Species / Q000378:metabolism; D020545:Respiratory Mucosa / Q000166:cytology* / Q000187:drug effects; D024483:Vitamin K 3 / Q000633:toxicity*
D006873:Hydroquinones; D009285:Naphthoquinones; D017382:Reactive Oxygen Species; D019328:Buthionine Sulfoximine; C063002:2,3-dimethoxy-1,4-naphthoquinone; D024483:Vitamin K 3; D006861:Hydrogen Peroxide