3-溴丙酮酸对肝癌Bel7402细胞凋亡及相关蛋白表达的影响

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

    • 摘要: 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。

       

      Abstract: 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.

       

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