抗增殖蛋白过表达对HepG_2细胞增殖和凋亡的影响

Effect of prohibitin over-expression on proliferation and apoptosis of human hepatocellular carcinoma cell line HepG2 cells

目的探讨抗增殖蛋白(PHB)过表达对人肝癌细胞系HepG_2细胞增殖和凋亡的影响。方法在已构建的PHB真核表达质粒pEGFP-N1-PHB瞬时转染HepG_2细胞,采用四甲基偶氮唑盐比色法(MTT法)检测HepG_2细胞增殖;使用流式细胞仪检测HepG_2细胞周期和凋亡。结果转染p EGFP-N1-PHB细胞增殖明显低于pEGFP-N1空载转染细胞或未转染细胞(P<0.05);在转染后48 h,转染组G2/M期细胞比例为(27.84±0.47)%,显著高于空载转染组的(17.21±0.64)%或未转染组的(22.67±0.33)%(P<0.001);转染组细胞凋亡率为(31.72±0.35)%,显著高于空载转染组的(18.66±0.56)%或未转染组的(13.47±0.94)%(P<0.001)。结论 PHB过表达可抑制人肝癌HepG_2细胞增殖,诱导细胞进入G2/M期后被阻滞,并诱导细胞凋亡。

Objective To investigate the effect of prohibitin （PHB） over-expression on proliferation and apoptosis of human hepatocellular carcinoma cell line HepG2 cells. Methods The eukaryotic expression plasmid （pEGFP-N1-PHB） had been constructed and the HepG2 cells were transiently transfeeted with recombinant plasmid. Measurement of cell proliferation was performed by using MTT assay. Flow cytometry was carried out to detect cell cycle alterations and apoptosis. Results The cell growth curves showed that the proliferation of HepG2 cells were significantly decreased after transfection with pEGFP-N1-PHB as compared with pEGFP-N1 or controls （P〈0.05）;At 48 hours after transfection,the proportion of G2/M phase of HepG2 cells in pEGFP-N1- PHB-transfeeted group was （27.84±0.47）% ,significantly higher than in pEGFP-Nl-transfected group [（17.21±0.64）%] or in the non-transfected group [（22.67±0.33）%,P〈0.001];The percentage of apoptotie cells in pEGFP- N1-PHB-transfeeted group was （31.72±0.35）%,much higher than （18.66±0.56） % in pEGFP-N1-transfeeted or non-transfected cells [（13.47±0.94）%,P〈0.001]. Conclusion PHB overexpression might inhibit the proliferation, induce G2/M phase arrest and apoptosis of HepG2 cells.