VEGF基因沉默对胰腺癌细胞化疗敏感性和Akt信号通路的影响  

Effects of VEGF down regulation by small interfering RNA on chemosensitivity and Akt signal pathway of human pancreatic cancer cell

作　　者：王晓燕[1] 周永静[1] 龚丹丹[1] 徐军[1] 徐岷[2] 范钰[1] 

机构地区：[1]江苏大学附属人民医院肿瘤研究所,江苏镇江212002 [2]江苏大学附属医院消化科

出　　处：《中华胰腺病杂志》2011年第2期95-97,共3页Chinese Journal of Pancreatology

基　　金：镇江市社会发展基金（SH2009014）;镇江市卫生事业发展基金（WS0908）

摘　　要：目的 观察靶向血管内皮生长因子（vascular endothelial growth factor,VEGF）基因小干扰RNA（small interfering RNA,siRNA）对胰腺癌BxPC3细胞化疗药物敏感性的影响,并探讨其可能机制.方法 将培养后的BxPC3细胞分为单纯BxPC3细胞组、脂质体处理组、错配siRNA转染组和VEGFsiRNA转染组.以5、10、20、100、200 nmol/L的VEGF siRNA转染BxPC3细胞.采用实时定量RT-PCR和ELASA法检测细胞VEGF mRNA和蛋白的表达;MTT法检测吉西他滨对各组细胞生长的影响;蛋白质印迹法检测BxPC3细胞Akt蛋白的磷酸化.结果 VEGF siRNA转染后,BxPC3细胞VEGF mRNA和蛋白呈浓度和时间依赖性明显下调,但对BxPC3细胞的增殖无明显影响.用0.2 μmol/L吉西他滨处理细胞48 h后,BxPC3细胞组、错配siRNA组及5、10、20 nmol/L siRNA组细胞的生长抑制率分别为（16.9±0.3）%、（17.3±0.3）%、（28.8±0.4）%、（52.2±0.3）%、（75.4±0.4）%,抑制率与siRNA浓度相关（r=0.928）.同时,siRNA转染后细胞Akt蛋白的磷酸化被明显抑制.结论 VEGF基因在胰腺癌BxPC3细胞耐药中起着重要的作用,其机制可能与Akt蛋白磷酸化有关.Objective To study the effects VEGF small interfering RNA （siRNA） on chemosinsitivity of human BxPC3 cell and its mechanism. Methods BxPC3 cells were divided into single BxPC3 cell group,lipofection group, scrambled siRNA transfection （200 nmol/L） group, VEGF siRNA transfection group.VEGF siRNA （5, 10, 20, 100,200 nmol/L） was used to transfect BxPC3 cells. Expressions of VEGF mRNA and protein were determined by real-time PGR and ELISA assay, respectively. MTT was performed to examine the inhibitory effects of gemcitabine on BxPC3 cells of each group. The phosphorylated-Akt protein was evaluated by Western blotting. Results After VEGF siRNA transfection, the expression of VEGF mRNA and protein in BxPC3 cells was down-regulated in a dose-and time-dependent manner, but there was no effect on BxPC3 cells proliferation. After 0. 2 μmol/L of gemcitabine treatment for 48 h, the inhibitory rates were （ 16.9 ±0.3）%, （17.3 ±0.3）%, （28.8 ±0.4）%, （52.2 ±0.3）%, （75.4 ±0.4）% in BxPC3 cell group,lipofection group, 5,10,20 nmol/L VEGF siRNA transfection group, and the inhibitory effects were correlated with siRNA concentration （ r = 0. 928 ）. The phosphorylated-Akt protein was reduced significantly in siRNA transfected cells. Conclusions VEGF gene plays an important role in BxPC3 cells resistence to chemotherapy through inhibiting Akt phosphorylation.

关 键 词：胰腺肿瘤 血管内皮生长因子 RNA 小分子干扰 化疗敏感性 AKT 

分 类 号：R735.9[医药卫生—肿瘤]