In vitro study of the effect of metabolism enzymes on benzo(a)pyrene-induced DNA damage in the scallop Chlamys farreri.

Acute toxicity effect of benzo(a)pyrene (BaP) on isolated scallop (Chlamys farreri) digestive gland cells was studied and a dose-dependent increase in toxicity was observed. The 8 μg/L of BaP had a significant toxic effect on isolated cells (p<0.05). In order to study the mechanism of CYP450, GST, SOD and MXR transporters involved in the production of DNA strand breakage such as DNA adduct formation and oxidative DNA damage by BaP were investigated in isolated digestive gland cells. Isolated cells were exposed in vitro to 0.8 μg/L of BaP for 24h in the dark at 25 °C in the absence or presence of cytochrome P450 inhibitor, GST inhibitor, Pgp inhibitor and antioxidant enzyme inhibitor. DNA adduct and 8-OHdG content were measured using the Enzyme-linked Immunosorbent Assay. The result indicated that DNA strand breakage was increased to 2 times compared with the control in the 0.8 μg/L of BaP treatment groups. The BaP-induced DNA adduct and 8-OHdG content increased significantly by inhibiting GST, while only 8-OHdG increased significantly when SOD was inhibited. The content of DNA adduct and 8-OHdG had no significant change when CYP450 was inhibited, while it decreased significantly when MXR transporters were inhibited. The result proved that GST play a key role in eliminating the BaP-induced DNA adduct and 8-OHdG, and SOD also had an important function in reducing the production of BaP-induced 8-OHdG.