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目的构建RNA i表达载体,并分析其对胰腺癌细胞PANC-1 surv iv in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surv iv in的表达,并把surv iv in基因克隆到T载体进行测序,构建针对surv iv in基因的干扰载体,用脂质体转染胰腺癌细胞PANC-1,RT-PCR、w estern-b lot分析干扰载体对surv iv in表达的抑制情况,流式细胞仪检测细胞凋亡。结果surv iv in在胰腺癌细胞PANC-1中高表达,其基因序列与G enB ank中公布的一致,干扰载体pTZU 6+1-svv2可以有效的抑制surv iv in的表达,抑制率约为74%,并诱导了肿瘤细胞的凋亡。结论用RNA i表达载体可以有效抑制胰腺癌细胞PANC-1中凋亡抑制蛋白surv iv in的表达并诱导胰腺癌细胞凋亡。
Objective To construct RNAi expression vector and analyze its inhibitory effect on the expression of PANC-1 survivin in pancreatic cancer cells. Methods The expression of survivin protein was detected by immunofluorescence and RT-PCR in pancreatic cancer cell line PANC-1. The survivin gene was cloned into T vector and sequenced. The interference vectors targeting surviv in gene were constructed, PANC-1 was transfected into pancreatic cancer cells by lipofectamine 2000. The inhibition of survivin expression by RT-PCR and western-b lot was analyzed by flow cytometry. Results Survivin was highly expressed in pancreatic cancer cell line PANC-1 and its sequence was consistent with that reported in GenBank. The interference vector pTZU 6 + 1-svv2 could effectively inhibit the expression of survivin with the inhibitory rate of about 74% , And induced tumor cell apoptosis. Conclusion RNAi expression vector can effectively inhibit the expression of survivin in pancreatic cancer cell PANC-1 and induce pancreatic cancer cell apoptosis.