c-Flip的基因表达在肝癌细胞对TRAIL诱导凋亡耐受中的作用  

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

作　　者：凌洪[1] 吕国悦[1,2] 郝恩源[1,2] 耿亚军[1,2] 陈儇[1,2] 

机构地区：[1]湖南衡阳南华大学附属第一医院急诊外科,衡阳421001 [2]吉林大学白求恩第一医院,长春130021

出　　处：《中国免疫学杂志》2011年第7期597-601,共5页Chinese Journal of Immunology

基　　金：国家自然科学基金(30901436);吉林省科技厅青年基金(20080162)资助

摘　　要：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。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 sensitivity of HepG2 and Hep3B to TRAIL was regained after c-FlipsiRNA transfection.The cell viability of HepG2 and Hep3B were 57.34%±6.13% and 43.98%±4.54% in siRNA groups,there was a significant difference compared with 96.44%±5.43% and 101.85%±6.41% in control groups（P〈0.01）.The SubG1 percent of cell cycle were 74.68%±6.95% and 78.34%±7.64%,also showed have a significant difference compared with 0.78%±0.07% and 3.62%±0.38%（P〈0.01）.The split of conducting protein Caspase-8,Caspase-3 in apoptosis conducting path of HepG2siRNA and Hep3BsiRNA could be induced alone by TRAIL.Conclusion：Tolerance of TRAIL induced apoptosis in hepatocellular carcinoma cell was associated with the expression of TIP protein in its conducting path,the split of apoptosis conducing protein Caspase-8,Caspase-3 can be induced by inhibiting the expression of c-Flip protein,which can accelerate apoptosis and recover the sensitivity of TRAIL induced apoptosis in hepatocellular carcinoma cell.

关 键 词：肿瘤坏死因子相关凋亡诱导配体 肝细胞肝癌 细胞内抑制蛋白 凋亡 

分 类 号：R34[医药卫生—基础医学]