Role of NAD(P)H:quinone oxidoreductase 1 (DT diaphorase) in protection against quinone toxicity

Abstract

NQO1−/− mice, along with Chinese hamster ovary (CHO) cells, were used to determine the in vivo role of NAD(P)H:quinone oxidoreductase 1 (NQO1) in cellular protection against quinone cytotoxicity, membrane damage, DNA damage, and carcinogenicity. CHO cells permanently expressing various levels of cDNA-derived P450 reductase and NQO1 were produced. Treatment of CHO cells overexpressing P450 reductase with menadione, benzo[ a]pyrene-3,6-quinone (BPQ), and benzoquinone led to increased cytotoxicity as compared with CHO cells expressing endogenous P450 reductase. In a similar experiment, overexpression of NQO1 significantly protected CHO cells against the cytotoxicity of these quinones. Knockout ( NQO1−/−) mice deficient in NQO1 protein and activity had been generated previously in our laboratory and were used in the present studies. Wild-type ( NQO1+/+) and knockout ( NQO1−/−) mice were given i.p. injections of menadione and BPQ, followed by analysis of membrane damage and DNA damage. Both menadione and BPQ induced lipid peroxidation in hepatic and non-hepatic tissues, indicating increased membrane damage. Exposure to BPQ also resulted in increased hepatic DNA adducts in NQO1−/− mice as compared with NQO1+/+ mice. The skin application of BPQ alone and BPQ + 12- O-tetradecanoylphorbol-13-acetate (TPA) failed to induce papillomas, or other lesions, for up to 50 weeks in either NQO1+/+ or NQO1−/− mice. The various results from CHO cells and NQO1−/− mice indicated that NQO1 protects against quinone-induced cytotoxicity, as well as DNA and membrane damage. The absence of BPQ-induced skin carcinogenicity in NQO1−/− mice may be related to the strain (C57BL/6) of mice used in the present study and/or due to poor BPQ absorption into the skin and/or due to detoxification of BPQ by cytosolic NRH:quinone oxidoreductase 2 (NQO2).