siRNA靶向沉默成纤维细胞激活蛋白α对胶质瘤细胞U87增殖的影响  

Effect of siRNA on proliferation of glioma cell line U87 by targeting fibroblast activation protein alpha

作　　者：陈渊[1] 王伟[1] 涂明[1] 吴哲褒[1] 郑伟明[1] 

机构地区：[1]温州医科大学附属第一医院,神经外科,浙江温州325015

出　　处：《温州医学院学报》2014年第4期241-244,共4页Journal of Wenzhou Medical College

基　　金：浙江省外科学重中之重学科开放基金资助项目(kfjj2011006)

摘　　要：目的 :探讨小分子干扰RNA(siRNA)靶向沉默成纤维细胞激活蛋白α(FAP-α)表达对神经胶质瘤细胞株U87增殖的影响。方法:实验分为空白对照组(Blank组)、阴性对照组(NC组)和siRNA组,通过脂质体将siRNA转染至胶质瘤细胞株U87,使用Western blot法检测FAP-α基因的蛋白水平,同时检测NC组和siRNA组Bcl-2、caspase-3的表达变化,使用Real-time PCR法检测FAP-α和增殖核抗原(PCNA)的mRNA水平,CCK-8法评价转染后NC组和siRNA组细胞的增殖能力。结果:①转染48 h后,siRNA组细胞FAP-αmRNA、蛋白的表达明显低于NC组和Blank组,差异有统计学意义(P<0.01),siRNA组较NC组caspase-3蛋白表达增加(P<0.05);Bcl-2蛋白表达明显降低(P<0.01),siRNA组较NC组PCNA mRNA表达明显下降(P<0.01)。②转染后48、72和96 h siRNA组U87细胞吸光度值低于NC组,差异有统计学意义(P<0.01)。结论:应用siRNA沉默FAP-α基因的表达,可以抑制胶质瘤细胞U87的增殖,诱导胶质瘤细胞的凋亡。Objective： To explore the effect of small interfering RNA （siRNA） -mediated fibroblast activa- tion protein alpha knock-down on proliferation of glioma cell line U87. Methods： The experiment is divided into Blank, NC and siRNA. Negative control or siRNA were transfected into U87cells by Lipofectamine 2000. After transfection, protein level of target gene FAP- α Bcl-2 and caspase-3 were detected by Western blot, mRNA level of PCNA and FAP-α were detected by Real-time PCR. The proliferation ability of NC group and siRNA group of cells were analyzed by CCK8 assay. Results： ① Forty-eighth after transfection, mRNA and protein expressions of FAP-α in siRNA group were remarkably reduced as compared with those in NC and Blank group （P〈0.05）; Compared with NC group, the expression of caspase-3 was higher （P〈0.05）, while that of Bcl- 2 was lower （P〈0.01）. The expression of PCNA mRNA was lower than NC group （P〈0.01）. ②The OD450 value of siRNA group was significantly lower than that in the NC group 24, 48, 72 and 96 h after transfection （P〈0.05）. Conclusion： siRNA-mediated fibroblast activation protein alpha knock-down can inhibit the proliferation of U87 glioma cells and promote apoptosis of glioma cells.

关 键 词：神经胶质瘤 小分子干扰RNA 成纤维细胞激活蛋白α 细胞增殖 

分 类 号：R739.41[医药卫生—肿瘤]