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、200μmol/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、85.0μmol/L。3-溴丙酮酸可诱导Bel7402细胞发生明显的细胞凋亡,50、100、200μmol/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。
Objective： To investigate the effect of 3-bromopyruvate on the apoptosis of liver carcinoma cells, and its underlying mechanisms. Methods ：The proliferative inhibition of Be17402 cells treated with 3-bromopyrnvate was measured by MqT assay. The cells were treated with different concentrations of 3-bromopyrnvate（ 0,50,100,200 μmol/L ）, and apoptosis was analyzed by flow eytometry 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 Be17402 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 Be17402 cells. The apoptotic rate of 50,100,200 μmol/L of 3-bromopyruvate treatment for 24 h was siguificantly different from that of the control group（ P 〈 0.01 ）. At the same time, 3-bromopyruvate significantly decreased the intracellular ATP level （ P 〈 O. 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 Be17402 cells. Conclusions：3-bromopyruvate can induce apoptosis in Be17402 cells, which may be correlated to the inadequate ATP supply, down-regulation of Bel-2, and up-regulation of Bax.
作者
潘琼
任凯
崇殿龙
张智瑞
周灿
刘浩
赵素容
PAN Qiong REN Kai CHONG Dian-long ZHANG Zhi-rui ZHOU Can LIU Hao ZHAO Su-rong(Faculty of Pharmacy,Bengbu Medical College ,Anhui Engineering Technology Research Center of Biochemical Pharmaceuticals, Bengbu Anhui 233030, Chin)
出处
《蚌埠医学院学报》
CAS
2016年第9期1129-1131,1135,共4页
Journal of Bengbu Medical College
基金
国家自然科学基金项目(81372899)
安徽省高校自然科学研究重大项目(KJ2016A486)
安徽省大学生创新创业训练项目(201510367035)
蚌埠医学院科研基金项目(BYKY1408ZD)
