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.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
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)
