胰腺癌细胞survivin表达RNAi抑制载体的构建及其抑制作用的研究

目的构建胰腺癌细胞生存蛋白（survivin）表达的RNA干扰（RNAi）抑制载体，并研究其对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survivin及其mRNA的表达，并把survivin基因克隆到T载体进行测序；构建针对survivin缺失碱基基因和survivin基因的RNAi载体si-svv-1和si—SVV-2，用脂质体转染胰腺癌细胞PANC-1后，用RT—PCR、DNA梯状电泳及流式细胞术分析比较2种干扰载体对survivin mRNA表达的抑制情况和检测细胞凋亡。结果survivin在胰腺癌细胞PANC-1中高表达；si—svv-2对survivin mRNA的表达抑制率达（72．43±8．04）％，而si—svv-1为0；si—svv-2能诱导胰腺癌细胞PANC-1的凋亡，72h细胞凋亡率达（12．36±1．44）％。结论本研究成功建立胰腺癌细胞survivin表达RNAi抑制载体，该载体可特异有效地抑制胰腺癌细胞PANC-1survivin的表达并诱导胰腺癌细胞PANC-1凋亡。

Study of Inhibiting The Expression of Survivin in Pancreatic Cancer Cells with RNA Interference

Objective To investigate the inhibitory effects of RNA interference(RNAi)expression vector on the expression of survivin in pancreatic cancer cell PANC-1.Methods The protein and mRNA expressions of survivin were examined with immunofluorescence and RT-PCR.The survivin gene was cloned into the T-vector and sequenced.The RNAi expression vectors targeting survivin,named si-svv-1 and si-svv-2 respectively according to whether they harbored a mutation or no mutation,were constructed and transfected into PANC-1 c...