RNA干扰抑制survivin表达并诱导胰腺癌细胞凋亡的实验研究
[The experiment study on RNAi inhibiting the expression of survivin and inducing the apoptosis of pancreatic cancer cells].
作者信息
He Man-xi, Meng Ling-xiang, Tang Yong, Li Jian-shui, Tian Bo-le, Hu Wei-ming, Huang Xiong, Liu Zhong-rong, Zhang Zhao-da
机构信息
Department of General Surgery, West China Hospital, Sichuan University, Chengdu 610041, China.
出版信息
Sichuan Da Xue Xue Bao Yi Xue Ban. 2006 Jul;37(4):520-4.
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.
目的
探讨RNA干扰（RNAi）表达载体抑制生存素表达并诱导胰腺癌细胞PANC-1凋亡的作用。
方法
采用逆转录-聚合酶链反应（RT-PCR）和免疫荧光法检测生存素的表达。将生存素基因克隆到T载体并测序，同时成功构建针对生存素的RNAi表达载体，然后用脂质体转染PANC-1细胞。采用RT-PCR和蛋白质印迹法检测生存素mRNA的表达。用流式细胞术检测细胞凋亡率。
结果
PANC-1细胞中生存素呈高表达。DNA序列与基因库中的生存素基因一致。pTZU6 + 1-svv2 RNAi表达载体可使生存素表达抑制约70%，凋亡细胞增加。
结论
RNAi表达载体可有效抑制PANC-1细胞中生存素的表达并诱导其凋亡。
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