c-Flip的基因表达在肝癌细胞对TRAIL诱导凋亡耐受中的作用

目的:观察c-Flip基因的表达变化在肝癌细胞对TRAIL诱导凋亡耐受中的作用。方法:以肝癌细胞HepG2、Hep3B为研究对象,制备c-FlipsiRNA的细胞模型,应用流式细胞仪分析两种细胞模型对TRAIL作用后细胞周期的变化,Western blot检测其在c-Flip基因沉默前后凋亡信号传导蛋白Caspase-8、Caspase-3等的表达变化。结果:转染c-FlipsiRNA后,细胞恢复了对TRAIL诱导凋亡的敏感性,当TRAIL浓度为100 ng/ml时,HepG2及Hep3B转染组的细胞生存率为57.34%±6.13%和43.98%±4.54%,显著低于对照组的96.44%±5.43%和101.85%±6.41%(P<0.01);细胞生存周期SubG1期为74.68%±6.95%和78.34%±7.64%,显著高于对照组的0.78%±0.07%和3.62%±0.38%(P<0.01),转染组TRAIL单独作用,可以引起HepG2siRNA及Hep3BsiRNA凋亡传导通路上传导蛋白Caspase-8、Caspase-3的裂解。结论:肝癌细胞对TRAIL诱导凋亡的耐受可能与其传导通路上c-Flip蛋白的表达相关,抑制其表达可以导致凋亡传导蛋白Caspase-8、Caspase-3的裂解,促使凋亡的发生,恢复肝癌细胞对TRAIL诱导凋亡的敏感性。

The experimental study of c-Flip gene expression in TRAIL-induced apoptosis tolerance in hepatocellular carcinoma cell

Objective:To discuss the effect of c-Flip gene expression in TRAIL-induced apoptosis tolerance in hepatocellular carcinoma cell.Methods:We set hepatocellular carcinoma cell HepG2 and Hep3B as targets and prepared cells model of c-FlipsiRNA,cell cycle variation of the two types of cell models were analysed by Flowcytometry after being treated by TRAIL,the expression of signal transduction protein Caspase-8,Caspase-3,DFF-45 were detected by Western blot before and after TIP gene silencing.Results:The sensitiv...