Effects of 3-bromopyruvate on apoptosis and the expression of apoptosis-related proteins in liver carcinoma Bel7402 cells

Objective: To investigate the effect of 3-bromopyruvate on the apoptosis of liver carcinoma cells,and its underlying mechanisms.Methods: The proliferative inhibition of Bel7402 cells treated with 3-bromopyruvate was measured by MTT assay.The cells were treated with different concentrations of 3-bromopyruvate(0,50,100,200 μmol/L),and apoptosis was analyzed by flow cytometry with PI staining.The intracellular ATP level was detected by commercial kit.The expression of Bcl-2 and Bax protein was analyzed by Western blot.Results: 3-bromopyruvate inhibited the proliferation of Bel7402 cells in a time and concentration dependent manner,with an IC₅₀ value of 422.9,143.9,85.0 μmol/L at 24,48 and 72 h,respectively.3-Bromopyruvate also induced obvious apoptosis in Bel7402 cells.The apoptotic rate of 50,100,200 μmol/L of 3-bromopyruvate treatment for 24 h was significantly different from that of the control group(P<0.01).At the same time,3-bromopyruvate significantly decreased the intracellular ATP level(P<0.01).The result of Western blot showed that 3-bromopyruvate down-regulated the expression of antiapoptotic protein Bcl-2,and up-regulated the expression of proapoptotic protein Bax in Bel7402 cells.Conclusions: 3-bromopyruvate can induce apoptosis in Bel7402 cells,which may be correlated to the inadequate ATP supply,down-regulation of Bcl-2,and up-regulation of Bax.