3-溴丙酮酸对肝癌Bel7402细胞凋亡及相关蛋白表达的影响

目的:观察糖酵解抑制剂3-溴丙酮酸对肝癌Bel7402细胞凋亡的影响，并探讨其作用机制。方法:采用MTT法检测3-溴丙酮酸对Bel7402细胞的增殖抑制效应，PI单染流式细胞术检测不同浓度3-溴丙酮酸（0、50、100、200 μmol/L）对Bel7402细胞凋亡的影响，ATP检测试剂盒分析细胞内ATP水平，Western blot检测凋亡相关蛋白Bcl-2、Bax的表达。结果:3-溴丙酮酸可浓度和时间依赖性地抑制Bel7402细胞的增殖（P<0.01），作用24、48、72 h的IC₅₀值分别为422.9、143.9、85.0 μmol/L。3-溴丙酮酸可诱导Bel7402细胞发生明显的细胞凋亡，50、100、200 μmol/L 3-溴丙酮酸作用24 h的细胞凋亡率均明显高于对照组（P<0.01）。ATP水平检测结果表明，3-溴丙酮酸可明显降低Bel7402细胞内的ATP水平（P<0.01）。Western blot结果显示，3-溴丙酮酸可下调凋亡抑制蛋白Bcl-2的表达，并上调凋亡诱导蛋白Bax的表达。结论:3-溴丙酮酸具有诱导肝癌Bel7402细胞凋亡的作用，其机制可能是通过引起细胞内ATP降低、调节细胞凋亡相关蛋白Bcl-2和Bax的表达。

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.