Effect of Dichloroacetonitrile on the Apoptosis of HepG2 Cells

Objectives  To investigate the effect of dichloroacetonitrile (DCAN) on apoptosis in HepG2 cells and its related regulatory mechanism.

Methods  DCAN was dissolved with dimethyl sulfoxide (DMSO), HepG2 cells were exposed to 50 μmol/L, 100 μmol/L, 200 μmol/L and 400 μmol/L DCAN for 24 h, and HepG2 cells treated with DMSO only were served as the control. Apoptosis was tested by Annexin V/PI double staining and determined by flow cytometry. The activity of Caspase 3 was measured by fluorometric assay. The expression of pro-Caspase and P53 protein was detected by western blotting.

Results  Compared with the control group, the proportion of apoptotic cells in the 400 μmol/L DCAN-treated group was increased (P<0.05), the enzyme activity of Caspase 3 was significantly increased in all DCAN-treated groups with the increase of exposure doses, and the expression of pro-Caspase and P53 protein were down-regulated in the 200 μmol/L and 400 μmol/L DCAN-treated groups (P<0.05).

Conclusion  High concentration of DCAN might induce the apoptosis of HepG2 cells by raising the activity of Caspase 3.