短发夹RNA靶向沉默TPX2基因促进人肺腺癌A549细胞凋亡及其相关机制  

TPX2 gene-targeted short hairpin RNA induces the apoptosis of human lung adenocarcinoma A549 cells and its related mechanism

作　　者：刘莹[1] 张徽[1] 唐熹[1] 

机构地区：[1]重庆医科大学分子医学与肿瘤研究中心病理学教研室,重庆400016

出　　处：《肿瘤》2011年第12期1055-1060,共6页Tumor

摘　　要：目的:探讨靶向Xklp2靶蛋白(targeting protein for Xenopus kinesin-like protein2,TPX2)基因的短发夹RNA(short hairpin RNA,shRNA)对肺腺癌A549细胞凋亡的影响及其可能机制。方法:构建靶向TPX2基因的shRNA重组载体,将其转染至肺腺癌A549细胞中,RT-PCR检测细胞中TPX2、Aurora-A、p53和Bcl-2mRNA的表达,蛋白质印迹法检测TPX2蛋白的表达,FCM检测细胞周期和细胞凋亡情况。结果:成功构建重组载体pMagic4.1-shRNA-TPX2。将pMagic4.1-shRNA-TPX2转染至A549细胞后,TPX2、Aurora-A和Bcl-2mRNA的表达水平明显下调,p53mRNA的表达水平明显上调,TPX2蛋白的表达水平明显下调,细胞凋亡率明显增加,细胞阻滞于S期,与空白对照组(未转染组)和阴性对照组(转染pMagic4.1-shRNA-NC)相比,差异均有统计学意义(P<0.05)。结论:靶向TPX2的shRNA能促进肺腺癌A549细胞的凋亡,其作用可能与上调p53表达和下调Bcl-2表达有关。Objective： To investigate the effect of short hairpin RNA （shRNA） targeting Xenopus kinesin-like protein 2 （TPX2） gene on the apoptosis of human lung adenocarcinoma A549 cells and to explore its possible mechanism. Methods： The combination vector of shRNA targeting TPX2 gene was established and then it was transfected into A549 cells. The expression levels of TPX2, Aurora-A, p53 and Bcl-2 mRNAs in A549 cells after transfection with combination vector were detected by RT-PCR, and the expression level of TPX2 protein was determined by Western-blotting. The cell cycle distribution and the apoptosis were analyzed by flow cytometry （FCM）. Results： The combination vector of pMagic4.1- shRNA-TPX2 was successfully established. The expression levels of TPX2, Aurora-A and Bcl-2 mRNAs were down-regulated whereas the expression level of p53 mRNA was up-regulated in A549 cells after transfection with pMagic4.1-shRNA-TPX2, and the expression level of TPX2 protein was down-regulated. The apoptosis rate of A549 cells was increased after transfection with pMagic4.1-shRNA-TPX2, and the cell cycle was arrested at S phase. These changes in the pMagic4.1-shRNA-TPX2-transfected group were significantly different from those in the blank control group （without transfection with combination vector） and the negative control group （transfection with pMagic4.1-shRNA-NC） （P〈0.05）. Conclusion： The shRNA targeting TPX2 gene can induce the apoptosis of A549 cells, and this effect may be related to up-regulation of p53 expression and down-regulation of Bcl-2 expression.

关 键 词：肺肿瘤 RNA干扰 细胞凋亡 基因 TPX2 

分 类 号：R734.2[医药卫生—肿瘤]