Survivin基因沉默对胰腺癌细胞Patu8988增殖及其对吉西他滨敏感性的影响

目的研究Survivin基因沉默对人胰腺癌Patu8988细胞增殖和对化疗药物吉西他滨敏感性的影响。方法设计合成Survivin的siRNA序列，Lipofectamine^TM 2000转染入Patu8988细胞。采用RT．PCR和Western blot检测Survivin在干扰后mRNA和蛋白的表达情况，利用流式细胞仪检测细胞周期。通过MTY法和细胞克隆形成试验法观察Survivin基因沉默后Patu8988细胞对吉西他滨的敏感性。结果Survivin基因沉默48h后，Patu8988细胞的Survivin基因和蛋白表达明显降低，基因表达分别为：对照组：0．78±0．03，空白组：0．82±0．06，实验组：0．52±0．05；蛋白表达分别为：对照组：0．77±0．21，空白组：0．77±0．26，实验组：0．57±0．03，差异有统计学意义（P〈0．05）。细胞周期被阻滞在G0／G1期，S期细胞数减少，其差异有统计学意义（P〈0．05）。Survivin基因沉默组细胞对吉西他滨的敏感性显著性增强。结论Survivin特异性siRNA能显著沉默Patu8988细胞Survivin基因，抑制细胞增殖，并增强Patu8988细胞对吉西他滨的敏感性。

Survivin gene silencing inhibited proliferation of human pancreatic cancer cell Patu8988 and enhanced their sensitivity to Gemcitabi

Objective To investigate the effect of small interfering RNA （siRNA）-mediated Sur- vivin knock-down on proliferation of human pancreatic cancer cell line Patu8988 and their sensitivity to Gemcitabi. Methods The siRNA against Survivin was constructed and transfected into Patu8988cells with LipofectamineTM 2000. The expression of Survivin was detected by RT-PCR and Western blot. Flow cytome- try was used to detect the cell cycle. Sensitivity to Gemcitabi after transfection were examined by MTY and clonogenic assay. Results In Patu8988 cells, the protein and mRNA levels of Survivin were decreased sig- nificantly after transfeetion, gene expression： control group： 0. 78 ＋ 0. 03, blank control group： 0. 82 ~ 0. 06, experimental group： 0. 52 ~0.05; protein expression were as follows： control group： 0. 77 -＋0. 21, blank control group ： 0. 77 ~ 0. 26, experimental group ： 0. 57 ＋ 0. 03, and the difference was statistically significant （ P 〈 0. 05 ）. And reduction of proliferation was related to an increase in the fraction of G0/G~ phase. The sensitivity of Patu8988 ceils to Gemcitabi was increased significantly after transfeetion. Conclu- sions The Survivin special siRNA silenced Survivin, decreased Patu8988 ceils proliferation and enhanced their sensitivity to Gemcitabi.