Detection of benzo(a)pyrene diol epoxide-DNA adducts in mononuclear white blood cells by CE immunoassay and its application to studying the effect of glutathione depletion.
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| Abstract |
High levels of benzo(a)pyrene diol epoxide (BPDE)-DNA adducts in white blood cells have been indicated as a risk factor for lung cancer. Sensitive, specific, fast and cost-efficient techniques for the detection of BPDE-DNA adducts in white blood cells are required for routine human biomonitoring. In the present study, an immunoassay based on CE/LIF was developed for the detection of BPDE-DNA adducts in mononuclear white blood cells (MNCs). Although glutathione (GSH) conjugation catalyzed by glutathione-S-transferase (GST) is considered to be the major pathway for inactivating BPDE, the effect of GSH depletion on BPDE-DNA adduct formation in MNCs has not been assessed. Therefore, we applied the newly developed method to study the effect of GSH depletion by D,L-buthionine-[S,R]-sulfoximine (BSO) on the level of DNA adducts. We found that pretreatment of MNCs with 150 microM BSO for 2 h prior to BPDE exposure increased the level of BPDE-DNA adducts appreciably (by approximately 70%). Further investigations revealed that the 2-h BSO treatment neither decreased the GSH level instantly nor affected GST activity; rather, it prevented the induction of GSH in response to subsequent BPDE incubation. The blocked synthesis of GSH might be responsible for the elevated level of BPDE-DNA adducts in MNCs after BSO and BPDE treatment.
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| Authors | Adrienne LeBlanc, Shengwen Shen, Kristi Lew, Michael Weinfeld, X Chris Le |
| Journal | Electrophoresis (Electrophoresis) Vol. 30 Issue 9 Pg. 1558-63 (May 2009) ISSN: 1522-2683 [Electronic] Germany |
| PMID | 19343725 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't) |
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