论文部分内容阅读
目的探讨靶向survivin的siRNA联合肿瘤坏死因子诱导凋亡相关配体(tumor necrosis factor related apoptosis-inducing ligand,TRAIL)对肝癌细胞增殖与凋亡的影响。方法构建靶向survivin基因的siRNA真核表达载体,观察可溶性TRAIL对肝癌细胞增殖与凋亡的影响。结果采用RT-PCR检测显示,siRNA在mRNA水平抑制survivin基因表达为73％。MTT法检测到可溶性TRAIL对HepG2、HepG2／Silence(-)细胞增殖无明显的抑制作用(P>0．05),通过抑制survivin表达,可使HepG2／Silence(+)细胞的12 h存活率明显降低,48 h降至最低[HepG2／Silence(+)0．518±0．017,对照组0．741±0．005,P<0．01]。荧光显微镜检测HepG2／Silence(+)细胞12 h、24 h、48 h的细胞凋亡率分别为:11．85％±0．72％、28．97％±0．43％、41．80％±0．90％,与对照组相比差异有统计学意义(F=22．37,P<0．05)。结论靶向survivin的siRNA在抑制肝癌细胞中survivin表达的同时可增强TRAIL作用的敏感性。
Objective To investigate the effect of siRNA targeting survivin combined with tumor necrosis factor related apoptosis-inducing ligand (TRAIL) on the proliferation and apoptosis of hepatocellular carcinoma cells. Methods The eukaryotic expression vector targeting survivin gene was constructed and the effect of soluble TRAIL on the proliferation and apoptosis of hepatoma cells was observed. Results RT-PCR showed that siRNA inhibited the expression of survivin gene at mRNA level by 73%. MTT assay showed that soluble TRAIL had no significant inhibition on the proliferation of HepG2 and HepG2 / Silence (-) cells (P> 0.05). The survival rate of HepG2 / Silence (+) cells at 12 h was significantly decreased by inhibiting the expression of survivin Decreased 48 h to the lowest [HepG2 / Silence (+) 0.518 ± 0.017, control group 0.741 ± 0.005, P <0.01]. The apoptotic rates of HepG2 / Silence (+) cells at 12 h, 24 h and 48 h were 11.85% ± 0.72%, 28.97% ± 0.43% and 41.80% respectively by fluorescence microscopy. ± 0.90%, compared with the control group, the difference was statistically significant (F = 22.37, P <0.05). Conclusion siRNA targeting survivin can inhibit the expression of survivin in hepatocellular carcinoma cells and enhance the sensitivity of TRAIL.