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目的应用RNAi技术将c-FLIP(L)对应的SiRNA片段对大肠癌细胞株SW480凋亡的影响,明确RNAi靶向沉默c-FLIP基因在TRAIL介导的凋亡途径中的作用。方法与c-FLIP(L)对应的SiRNA片段转染入细胞,半定量RT-PCR法判断干扰前后FLIPmRNA水平的变化,Westernblot分析FLIP蛋白水平变化,比较分析干扰前后细胞对介导的凋亡敏感性的改变及转染前后TRAIL诱导的细胞凋亡的情况。结果 c-FLIP(L)可以减低SW480中FLIPmRNA和蛋白水平(P<0.01),转染FLIP-siRNA后,TRAIL对大肠癌胞株SW480的遏制作用显著加强(P<0.05),TRAIL介导凋亡明显增高(P<0.05)。结论 RNAi靶向沉默c-FLIP(L)并能提高SW480对TRAIL灵敏度并诱使SW480凋亡。
OBJECTIVE: To study the effect of SiRNA fragment of c-FLIP (L) on the apoptosis of colorectal cancer cell line SW480 by RNAi technique and to clarify the role of RNAi targeting silencing c-FLIP gene in TRAIL-mediated apoptosis pathway. Methods The siRNA fragment corresponding to c-FLIP (L) was transfected into cells. The changes of FLIP mRNA level before and after interference were determined by semi-quantitative RT-PCR. The levels of FLIP protein were analyzed by Western blot analysis. Sexual changes and TRAIL-induced apoptosis before and after transfection. Results The FLIP mRNA and protein levels in SW480 cells were down-regulated by c-FLIP (L) (P <0.01). After transfected with FLIP-siRNA, the suppressive effect of TRAIL on SW480 cells was significantly enhanced The death was significantly higher (P <0.05). Conclusion RNAi targeting silencing c-FLIP (L) can increase SW480 sensitivity to TRAIL and induce SW480 apoptosis.