RNA干扰抑制登革病毒复制的研究  

Study on the Inhibitory Effect of RNA Interference on Replication of Dengue Virus

作　　者：岳锦亚[1,2] 吴新伟[1] 伍业健[1] 李向忠[1] 蒋力云[1] 李巧艳[2] 李磊[2] 杨霞[2] 

机构地区：[1]广州市疾病预防控制中心,广州510080 [2]中山大学中山医学院,广州510080

出　　处：《病毒学报》2010年第5期373-378,共6页Chinese Journal of Virology

基　　金：广东省科技计划项目(2008B030303041);广州市科技计划项目(2006J1-C0141;2008J1-C191;2008Z1-E231);广州市医学科学技术研究重点项目(2006-ZDi-10;2008-ZDi-12)

摘　　要：为了研究RNA干扰(RNAi)对Ⅰ型登革病毒(DENV-1)在白纹伊蚊C6/36细胞内复制的影响,本研究设计并合成针对I型登革病毒Pr M基因的小干扰RNA,以脂质体法转染入C6/36细胞后,用DENV-1感染已转染的细胞,观察细胞病变效应,MTT法检测细胞存活率,荧光定量RT-PCR检测登革病毒RNA含量。结果表明:转染siRNA的C6/36细胞在受登革病毒攻击7天后仍无明显细胞病变效应,细胞存活率比对照组提高2.26倍,细胞内登革病毒RNA拷贝数比对照组降低约97.54%。说明利用RNA干扰技术能有效抑制登革病毒核酸在C6/36细胞内复制,并对细胞具有一定保护作用,为登革热的防治提供了新的思路。To investigate the inhibitory effect of RNA interference（RNAi） on dengue virus Ⅰ（DENV-1） replication.Small interfering RNA（siRNA） against the PreM gene of dengue virus was synthesized and transfected into C6/36 cells with liposome,which was then attacked by DENV-1 virus.The antiviral effect of siRNA was evaluated by cytopathic effect（CPE）,the cell survival rate measured by MTT,and virus RNA quantified by real-time RT-PCR.The results showed that after 7 days post infection of dengue virus,the transfected C6/36 cells showed less CPE.The cell survival rate of the transfected C6/36 cells increased by 2.26 fold,and the amount of virus RNA in the transfected cells was reduced by about 97.54% as well.These findings indicated that the siRNA could effectively inhibit dengue virus RNA replication,and protect C6/36 cells from viral attack,indicating its potential role in prevention and treatment of dengue fever.

关 键 词：登革病毒 C6/36细胞 RNA干扰 荧光定量PCR 

分 类 号：R373.2[医药卫生—病原生物学]