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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。
AIM: To investigate the effect of short hairpin RNA (shRNA) targeted to target protein for Xenopus kinesin-like protein 2 (TPX2) on the apoptosis of lung adenocarcinoma A549 cells and its possible mechanism. Methods: shRNA recombinant vector targeting TPX2 gene was constructed and transfected into lung adenocarcinoma A549 cells. The expressions of TPX2, Aurora-A, p53 and Bcl-2 mRNA were detected by RT-PCR. The expression of TPX2 protein FCM was used to detect cell cycle and apoptosis. Results: The recombinant vector pMagic4.1-shRNA-TPX2 was successfully constructed. After transfection of pMagic4.1-shRNA-TPX2 into A549 cells, the expression levels of TPX2, Aurora-A and Bcl-2 mRNA were down-regulated, the expression of p53 mRNA was up-regulated, the expression of TPX2 protein was down-regulated and the apoptosis rate was significantly Compared with the blank control group (untransfected group) and the negative control group (transfected with pMagic4.1-shRNA-NC), the difference was statistically significant (P <0.05). Conclusion: shRNA targeting TPX2 can promote the apoptosis of lung adenocarcinoma A549 cells, which may be related to the up-regulation of p53 and the down-regulation of Bcl-2 expression.