RNAi抑制胰腺癌细胞survivin表达并诱导细胞凋亡的实验研究

目的构建RNAi表达载体，并分析其对胰腺癌细胞PANC-1 survivin表达的抑制作用。方法用免疫荧光技术和RT-PCR检测胰腺癌细胞PANC-1中凋亡抑制蛋白survivin的表达，并把survivin基因克隆到T载体进行测序，构建针对survivin基因的干扰载体，用脂质体转染胰腺癌细胞PANC-1，RT-PCR、western-blot分析干扰载体对survivin表达的抑制情况，流式细胞仪检测细胞凋亡。结果survivin在胰腺癌细胞PANC-1中高表达，其基因序列与GenBank中公布的一致，干扰载体pTZU6＋1-svv2可以有效的抑制survivin的表达，抑制率约为74％，并诱导了肿瘤细胞的凋亡。结论用RNAi表达载体可以有效抑制胰腺癌细胞PANC-1中凋亡抑制蛋白survivin的表达并诱导胰腺癌细胞凋亡。

The Experiment Study on RNAi Inhibiting the Expression of Survivin and Inducing the Apoptosis of Pancreatic Cancer Cells

Objective To investigate the inhibitory effects of RNAi expression vector suppressing the expression of survivin and inducing the apoptosis of pancreatic cancer cell PANC-1.Methods The expression of survivin was examined with RT-PCR and immunofluorescence protocols.The survivin gene was cloned into the T-vector and sequenced,at the same time,the RNAi expression vectors aimed survivin were successfully constructed,and then transfected into PANC-1 cells with liposomes.The expression of survivin mRNA was detected with RT-PCR and Western-blot techniques.The rate of cell apoptosis was measured with FACS.Results There was a high degree expression of survivin in PANC-1 cells.The DNA sequence was according with the survivin gene in GENE BANK.The survivin expression was inhibited about 70% by pTZU6+1-svv2 RNAi expression vectors,and the apoptosis cells increased.Conclusion The RNAi expression vector can effectively inhibit the survivin expression and induce the apoptosis in PANC-1 cells.