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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。
OBJECTIVE: To study the effect of RNA interference on the expression of Aurora A gene in U251 glioma cells and to investigate its effect on the chemosensitivity of U251 cells. Methods: The siRNA targeting 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 blot. MTT assay and flow cytometry Changes of Proliferation and Apoptosis of U251 Cells Transfected with Aurora A siRNA under the Action of Nimustine (ACNU). Results: After Aurora A siRNA transfection, the expression of Aurora A mRNA was inhibited in U251 cells (P <0.01), and the protein expression was decreased. ACNU significantly enhanced the proliferation of U251 cells transfected with Aurora A siRNA (P <0.01), and the apoptosis rate of U251 cells transfected with ACNU significantly increased (P <0.05). CONCLUSIONS: The specific siRNA against Aurora A gene synthesized in vitro successfully inhibited the expression of Aurora A gene in U251 cells and increased the chemosensitivity of glioma cells.