BaP Metabolism and DNA-adduct Formation in Cultured Human Lymphocytes TreatedIn Vitrowith BaP and (-)-BaP-7,8-dihydrodiol

Abstract

Nine samples of human lymphocytes from six healthy subjects were treated in vitro for 24 hours with [3H]-benzo[a]pyrene (BaP) and unlabelled (-)-BaP-7,8-dihydrodiol; 1μg/ml and 0.5μg/ml, respectively. The separation by HPLC of BaP-DNA adducts showed a significant formation of syn-BaPDE-dGuo adducts in five out of the nine human lymphocyte samples treated in vitro with [3H]-BaP. BaP-(7,10/8,9) tetrol and BaP-(7/8,9,10) tetrol derived from (+)-anti-BaPDE were the predominant isomers extracted from the culture medium of all the lymphocyte samples treated with (-)-BaP-7,8-dihydrodiol (mean ratio of anti-BaPDE/syn-BaPDE tetrols 5.6 + 1.2), suggesting that the metabolic activation of (-)-BaP-7,8-dihydrodiol occurred by its epoxidation to (+)-anti-BaPDE much more frequently (4–8 times) than by epoxidation to the (-)-syn-BaPDE form. In contrast to the predominant formation of anti-BaPDE tetrols, the analysis by HPLC of DNA adducts in the five subjects examined also revealed the significant formation of syn-BaPDE-DNA adducts (mean adduct ratio of (+)-anti-/(-)-syn-BaPDE-DNA = 2.6 ± 0.7). In six human lymphocyte samples treated in vitro with anti-BaPDE and syn-BaPDE, the anti-BaPDE bound to DNA 2–5 times more efficiently than did the syn-BaPDE. However, after 24 hours, the level of anti-BaPDE-DNA adducts significantly decreased almost to the level of the syn-BaPDE-DNA adducts, suggesting that a stereoselective repair mechanism could have preferentially removed the anti-BaPDE-DNA adducts.