敲低Aurora-A抑制HepG2人肝癌细胞增殖并促进其凋亡

Knockdown of Aurora-A inhibits proliferation and promotes apoptosis of HepG2 hepatocellular carcinoma cells

目的探讨敲低Aurora-A对HepG2人肝癌细胞增殖和凋亡的影响。方法设计Aurora-A短发夹RNA(shRNA)片段,构建和包装Aurora-A shRNA慢病毒载体,转染HepG2细胞,实时定量PCR检测Aurora-A mRNA表达,Westernblot法检测Aurora-A蛋白及磷酸化水平,噻唑蓝(MTT)法检测细胞增殖,流式细胞术检测细胞凋亡。结果成功构建Aurora-A shRNA慢病毒载体,转染后的HepG2细胞Aurora-A蛋白磷酸化水平明显降低。敲低Aurora-A后,HepG2细胞增殖显著降低,细胞凋亡率显著增加。结论敲低Aurora-A的表达抑制HepG2细胞增殖并促进其凋亡。

Objective To investigate the effects of knockdown of Aurora-A gene on the proliferation and apoptosis of HepG2 human hepatocellular carcinoma cells.Methods Aurora-A short hairpin RNA(Aurora-A shRNA)was designed and Aurora-A shRNA lentiviral vector was constructed and packed,and then transfected into HepG2 cells.Aurora-A mRNA expression was detected by real-time quantitative PCR.Aurora-A protein expression and phosphorylation level were detected by Western blotting.Cell proliferation was tested by MTT assay.Cell apoptosis was analyzed by flow cytometry.ResultsThe Aurora-A shRNA lentiviral vector was successfully constructed and Aurora-A protein phosphorylation level was significantly reduced in HepG2 cells transfected with the lentiviral vector.When Aurora-a was knocked down,the proliferation of HepG2 cells decreased and the apoptosis rate increased significantly.Conclusion Knockdown of Aurora-A can inhibit the proliferation and promote the apoptosis of HepG2 cells.