【摘要】 目的探讨小干扰RNA(siRNA)沉默胶质瘤细胞BRG1基因表达对其增殖的影响及其机制。方法化学合成BRG1 siRNA,脂质体介导转染胶质瘤U251和U87细胞。CCK-8细胞增殖实验检测细胞增殖,流式细胞仪分析细胞周期变化,Western blot检测BRG1、cyclin家族、CDK抑制剂蛋白表达水平。结果转染BRG1 siRNA可以有效减少胶质瘤U251和U87细胞BRG1表达,细胞增殖下降,G1期细胞增加。BRG1沉默后cyclin D1和cyclin B1表达降低,CDK抑制剂p21和p27没有明显变化。结论沉默胶质瘤细胞BRG1表达影响细胞周期蛋白使细胞停滞在G1期,最终抑制胶质瘤细胞增殖。更多还原

【Abstract】 Objective To investigate the effect of siRNA silencing BRG1 gene on glioma cell proliferation and its molecular mechanism. Methods Chemically synthesized small interfering RNA targeting BRG1 was transfected into human glioma cell lines U251 and U87 by siLentFect Lipid Reagent. The proliferation of both glioma cells after BRG1 interference was measured by CCK-8 assay. The cell cycles of glioma cells were detected by flow cytometry.The expression levels of BRG1, cyclin family and CDK inhibitor proteins were detected by western blot. Results BRG1 interference could drastically decrease the ability of cell proliferation in both glioma cells. The G1 population in both glioma cells treated by BRG1 siRNA was increased significantly than that in the cells treated by control siRNA. The Western blot results showed that BRG1 knock-down inhibited expression of both cyclin D1 and cyclin B1, but the expression of p21 and p27. Conclusion BRG1 siRNA could effectively decrease the expression of BRG1 in glioma cell and arrest cell cycle at G1 phase and significantly inhibit the cell proliferation.更多还原