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目的探讨RNA干扰技术抑制人胶质瘤U251细胞中Aurora A基因的表达,研究其对U251细胞增殖及凋亡的影响。方法设计并合成特异性针对Aurora A基因的siRNA,转染至U251细胞中,采用RT-PCR和Western blotting检测Aurora A mRNA和蛋白的表达情况,同时利用四甲基偶氮唑盐(MTT)实验和流式细胞仪观察转染后U251细胞增殖抑制率和细胞凋亡,透射电镜观察细胞凋亡超微结构改变。结果转染Aurora AsiRNA后,U251细胞Aurora A mRNA表达受到抑制(P<0.01),蛋白表达水平降低;U251细胞增殖的抑制率和细胞凋亡率显著增高(P<0.01),且U251细胞发生了显著的凋亡形态改变。结论体外合成的针对Aurora A基因的特异性siRNA成功地抑制了U251细胞中Aurora A基因的表达,并能抑制胶质瘤细胞增殖、促进凋亡,提示Aurora A可能成为胶质瘤基因治疗的新靶点。
Objective To investigate the effect of RNA interference on Aurora A gene expression in human U251 glioma cells and its effect on the proliferation and apoptosis of U251 cells. Methods siRNA specific to Aurora A gene was designed and synthesized and transfected into U251 cells. The expression of Aurora A mRNA and protein was detected by RT-PCR and Western blotting. MTT assay The proliferation inhibition rate and apoptosis of U251 cells were observed by flow cytometry. The ultrastructural changes of apoptosis were observed by transmission electron microscope. Results After Aurora AsiRNA transfection, the expression of Aurora A mRNA in U251 cells was inhibited (P <0.01), and the expression of Aurora A mRNA was decreased. The inhibition of U251 cells proliferation and apoptosis were significantly increased (P <0.01) Significant changes in apoptotic morphology. Conclusions The specific siRNA against Aurora A gene synthesized in vitro successfully inhibits the expression of Aurora A gene in U251 cells and inhibits the proliferation and promotes the apoptosis of glioma cells, suggesting Aurora A may be a new gene therapy for glioma Targets.