RNA干扰抑制Aurora A表达对胶质瘤U251细胞化疗敏感性的影响

目的:采用RNA干扰技术抑制人脑胶质瘤U251细胞中Aurora A基因的表达,研究其对U251细胞化疗敏感性的影响。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 blot检测Aurora A mRNA和蛋白表达情况,同时利用MTT试验和流式细胞仪检测在化疗药物尼莫司汀(ACNU)作用下转染Aurora A siRNA前后U251细胞增殖及凋亡情况的变化。结果:转染Aurora A siRNA后,U251细胞Aurora A mRNA表达受到抑制(P<0.01),蛋白表达水平降低。ACNU对转染Aurora A siRNA后U251细胞的增殖抑制作用明显增强(P<0.01),同时在ACNU作用下转染后U251细胞的凋亡率也显著增加(P<0.05)。结论:体外合成的针对Aurora A基因的特异性siRNA成功的抑制了U251细胞中Aurora A基因的表达,并能提高胶质瘤细胞的化疗敏感性。

Effects of Aurora A Inhibition by RNA Interference on Chemotherapy Sensitivity of U251 Cells

Objective: To study the inhibitory effect of RNA interference on the expression of Aurora A in U251cells,and the influence on chemotherapy sensitivity of U251 cells.Methods: The siRNA specific for Aurora A was synthesized and transfected into U251 cells.Aurora A mRNA expression and protein contents were detected by RT-PCR and Western blot respectively.Under the action of chemotherapy drug Nimustine,the changes in proliferation and apoptosis of U251 cells before and after the transfection of Aurora A siRNA were studied by MTT and FCM.Results: The expression level of Aurora A mRNA was significantly decreased(P<0.01),and the protein content of Aurora A was also obviously reduced.The inhibitory rate of U251 cells after transfected with Aurora A siRNA was enhanced significantly by Nimustine(P<0.01).While under the action of Nimustine,the apoptosis rate of U251 cells after transfection was also significantly increased(P<0.05).Conclusion: The expression of Aurora A gene can be inhibited by siRNA successfully,and it results in the improvement of the chemosensitivity of glioma cells in vitro.