CAJ | 학술논문

目的观察受体作用蛋白(RIP)基因的表达变化在肝癌细胞对肿瘤坏死因子相关凋亡诱导配体(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RIP siRNA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RIP基因沉默前后凋亡信号传导蛋白Caspase-8、Caspase-3、DFF-45的表达变化.结果转染RIP siRNA后,细胞恢复了对TRAIL诱导凋亡的敏感性,当TRAIL浓度为100 μg/L时,HepG2及Hep3B转染组的细胞生存率为(60.02±5.23)％和(50.78±3.76)％,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(76.89±6.68)％和(72.45±7.31)％,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HepG2 siRNA及Hep3B siRNA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TRAI诱导凋亡的耐受可能与其传导通路上RIP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.
목적 관찰수체작용단백(RIP)기인적표체변화재간암세포대종류배사인자상관조망유도배체(TRAIL)유도조망내수중적작용.방법 이간암세포HepG2、Hep3B위연구대상,제비RIP siRNA적세포모형,응용류식세포의분석량충세포모형대TRAIL작용후세포주기적변화,Western blot검측기재RIP기인침묵전후조망신호전도단백Caspase-8、Caspase-3、DFF-45적표체변화.결과 전염RIP siRNA후,세포회복료대TRAIL유도조망적민감성,당TRAIL농도위100 μg/L시,HepG2급Hep3B전염조적세포생존솔위(60.02±5.23)％화(50.78±3.76)％,현저저우대조조적(96.44±5.43)％화(101.85±6.41)％(P＜0.01)；세포생존주기SubG1기위(76.89±6.68)％화(72.45±7.31)％,현저고우대조조적(0.78±0.07)％화(3.62±0.38)％(P＜0.01).전염조TRAIL단독작용,가이인기HepG2 siRNA급Hep3B siRNA조망전도통로상전도단백Caspase-8、Caspase-3적렬해.결론 간암세포대TRAI유도조망적내수가능여기전도통로상RIP단백적표체상관,억제기표체가이도치조망전도단백Caspase-8、Caspase-3적렬해,촉사조망적발생,회복간암세포대TRAIL유도조망적민감성.
Objective To discuss the role of receptor-interacting protein (RIP) gene expression in tumor necrosis factor-related apoptosis inducing ligand (TRAIL)-induced apoptosis tolerance in hepatocellular carcinoma cells.Methods By using hepatocellular carcinoma cell lines HepG2 and Hep3B as targets,cell models of RIP siRNA were prepared.The cell cycle variations of the two types of cell models treated with TRAIL were analyzed by using flow cytometry.The expression of signal transduction protein Caspase-8,Caspase-3 and DFF-45 was detected by using Western blotting before and after TIP gene ailencing.Results The sensitivity of HepG2 and Hep3B cells to TRAIL was regained after RIP siRNA transfection.The viability of HepG2 and Hep3B cells was (60.02 ±5.23)％ and (50.78 ±3.76)％ in siRNA groups,which was significantly lower than in control group [ (96.44 ± 5.43) ％ and ( 101.85 ± 6.41 ) ％ ]( P ＜ 0.01 ).The percentage of HepG2 and Hep3B cells in SubG1 of siRNA groups was (76.89 ± 6.68 ) ％and (72.45 ±7.31 )％ respectively,which was also significantly higher than in control group [ (0.78 ±0.07)％ and (3.62 ±0.38)％ ] (P ＜0.01 ).The split of transduction protein Caspase-8 and Caspase-3 in apoptosis transduction pathway of HepG2 siRNA and Hep3BsiRNA could be induced by TRAIL alone.Conclusion Tolerance of TRAIL-induced apoptosis in hepatocellular carcinoma cells was associated with the expression of TIP protein in its transduction pathway.The split of apoptosis transduction protein Caspase-8 and Caspase-3 can be induced by inhibiting the expression of RIP protein,which can accelerate apoptosis and recover the sensitivity of TRAIL-induced apoptosis in hepatocellular carcinoma cells.