特异性的VP1-siRNA对CVB3复制抑制作用的研究  

Inhibitory effect of specific siRNA on CVB3-VP1 expression in HeLa cells

作　　者：陈杰斌[1] 许从峰[2] 陈瑞珍[3] 虞勇[3] 李丽[1] 杜九中[1] 郭棋[3] 杨英珍[3] 邓晖[1] 李双杰[1] 

机构地区：[1]南华大学附属第一医院儿科,衡阳421001 [2]美国田纳西州立大学 [3]复旦大学中山医院卫生部病毒性心脏病重点实验室

出　　处：《中华微生物学和免疫学杂志》2006年第7期584-587,共4页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金(批准号:30271665)

摘　　要：目的研究特异性的CVB3-VP1 siRNA对CVB3体外复制的抑制作用的量效关系与时效关系。方法CVB3感染HeLa细胞,利用脂质体介导将CVB3-VP1 siRNA转染HeLa细胞,用RT-PCR法检测CVB3-VP1 RNA水平,免疫荧光检测CVB3-VPl蛋白的表达水平。结果60 pmol／L CVB3-VP1 siRNA是抑制CVB3 RNA及VP1表达的最佳剂量;在转染后的48 h,siRNA对CVB3-VP1的抑制作用达到最佳状态。结论特异性的CVB3-VP1 siRNA对CVB3-VP1蛋白表达水平及CVB3-RNA复制水平呈明显的剂量依赖关系和时效关系,转染后48 h呈现出完全抑制作用。Objective To investigate dose-dependence regulatory inhibition and time-effect relationship between CVB3-VP1 siRNA and the expression of CVB3-VP1 .in HeLa cells. Methods Four differents dose siRNA were transfected into HeLa cells by liposome （LipofectAMINE^TM2000）. 24, 48, 72, 96 h later, the CVB3-VP1 protein expression were detected by immunofluoreseence, and CVB3 mRNA level was tested by semi-quantitative RT-PCR. Results Observations have showed that 60 pmol/L CVB3-VP1 siRNA was the optimal dose, and the inhibitory effect on CVB3-VP1 expression was the optimum after transfection 48 h. Conclusion Specific siRNA have the dose-dependence and time-effect relationship to the expression level of CVB3-VP1. These results suggested a feasibility of using RNAi to elucidate pathogenesis of CVB3 myoearditis and that RNAi might represent a new approach for the treatment of CVB3 myoearditis.

关 键 词：小干扰RNA RNA干扰 柯萨奇B组病毒 衣壳蛋白 

分 类 号：R346[医药卫生—基础医学]