摘要
目的 拟筛选出能逆转大鼠胶质瘤多药耐药细胞株C6/VP16对依托泊苷耐药性的多药耐药相关蛋白1基因小干扰RNA(MRP1-siRNA)。方法 人工合成4对靶向MRP1的siRNA分别为siRNA1、siRNA2、siRNA3和siRNA4,以脂质体2000作为载体;以6-羧基荧光素标记转染siRNA的C6细胞评价转染效率;采用逆转录-PCR和免疫印迹分别检测MRP1mRNA和蛋白的表达。细胞计数盒-8试剂盒检测转染前后依托泊苷对肿瘤细胞的半生长抑制浓度(IC50)。结果 与siRNA1、siRNA4相比,siRNA2、siRNA3对MRP1基因的抑制作用更明显。转染siRNA2前后,依托泊苷对C6细胞的半生长抑制浓度分别为(0.873±0.0462)、(0.0927±0.039)μg/μl,两者相较差异显著(P<0.05)。结论 siRNA2、siRNA3可以有效抑制MRP1基因的表达,并能逆转肿瘤细胞对依托泊苷的耐药性。
Objective To screen a Muhidrug resistant-associated protein 1-small interference RNA （MRPI-siRNA） that may effectively reverse the etoposide （VP16） resistance of rat multidrug resistant glioma cell line C6 cells （C6/VP16）. Methods Four strands of siRNA targeting MRP1 were designed and synthesized. Lipofectamine 2 000 was used as a vector to transfect siRNA into rat glioma multidrug-resistant cell line C6 cells. The expressions of MRP1 mRNA and protein were determined respectively by reverse transcription-polymerase chain reaction and western blot. Cell counting kit-8 was applied to examination of the number of the glioma cells and the half maximal inhibitory concentration （IC50） value of vp16 to the cell line before and after the transfection. Results The inhibitory effects of siRNA2 and siRNA3 on the target （MMP1） gene were significantly stronger than those of siRNA1 and siRNA4 （P〈0.05）. IC50 of VP16 to C6 cells [（0.0927±0.039）ug/ul] was significantly lower after the siRNA2 transfection than that [（0.873±0.0462） ug/ul] before the siRNA2 transfection （P〈0.05）. Conclusion siRNA2 and siRNA3 can effectively inhibit MRP1 gene expression, and significantly reverse the resistance of C6 cells to VP16.
出处
《中国临床神经外科杂志》
2013年第3期154-157,共4页
Chinese Journal of Clinical Neurosurgery
