RNA干扰抑制胰腺癌细胞XIAP表达对化疗敏感性的影响

Suppression of pancreatic cancer cell eypression of XIAP by RNAi and its effect on sensitivily to chemotherapeutics

目的探讨运用载体介导的RNA干扰技术对X-连锁的凋亡抑制蛋白的抑制效率,以及观察抑制XIAP后胰腺癌细胞对化疗药物敏感性的变化。方法应用PBSHHI质粒构建针对XIAP的RNA干扰载体,转染胰腺癌细胞系SW1990;用RT-PCR和Westernblot检测XIAP的抑制效率,以gem-citabine处理细胞,流式细胞仪和荧光显微镜观察细胞凋亡的变化;分析XIAP的表达与细胞凋亡之间的关系。结果成功构建4个XIAP的RNA干扰载体,其中2个载体对XIAP的瞬时抑制效率达50%以上。XIAP被抑制后,细胞对化疗药物诱导的凋亡敏感性增强(P<0.05),XIAP的表达水平与细胞的凋亡指数之间呈负相关性(r=-0.809,P<0.05)。结论运用针对XIAP的RNA干扰载体可以有效地抑制XIAP的表达,提高胰腺癌细胞对化疗药物的敏感性。

Objective To investigate the possibility of XIAP inhibition by RNA interference （ RNAi ） vectors, and the chemotherapeutic sensitivity change of pancreatic carcinoma cells of SW1990 after XIAP inhibition. Methods RNA interference vectors against XIAP was constructed and transfected into the pancreatic carcinoma cell line SW 1990, the expression of XIAP was determined by RT-PCR and Western blot, while flow cytometry and Hoechst 33258 stain were employed to examine the apoptosis index of SW 1990 induced by gemcitabine. The relationship between XIAP and apoptosis index was analyzed by linear regression. Results Four RNAi vectors against XIAP were constructed, two of the four RNAi vectors could instantaneous inhibit XIAP expression more than 50%, The sensitivity of SW1990 to gemcitabine increased after MAP was inhibited （ P 〈 0. 05 ） , and the expression level of XIAP was negatively related to the apoptosis index （ P 〈 0. 05 ）. Conclusions RNAi vectors could effectively suppress the expression of XIAP, and enhance the sensitivety of pancreatic cancer cells to chemtherapentic drygs.