siRNA against the 2Apro genomic region of EV71 exerts potential antiviral effects
LIU Hai-bing1, KONG Zhen-zhen2, ZHAO Pei-pei2, MAO Ling-xiang3, SU Zhao-liang2, CHEN Jian-guo1
（1.Department of Clinical Laboratory, the Affiliated People′s Hospital of Jiangsu University, Zhenjiang Jiangsu 212002; 2.School of Medicine, Jiangsu University, Zhenjiang Jiangsu 212013; 3.Zhenjiang Center for Disease Control and Prevention, Zhenjiang Jiangsu 212004， China)
Abstract：Objective: To explore whether siRNA targeted the 2A protease genomic (2Apro) region of enterovirus 71 (EV71) possesses potential antiviral effects. Methods: RD cells were transfected with 40, 60 and 80 nmol/L BLOCKiT Fluorescent Oligo labeled with FITC, transfection efficiency of siRNA was detected by flow cytometry and fluorescence microscopy. RD cells were transfected with three siRNA2Apro (siRNA69, 294, 319), the experimental group, and siRNASCS,the negative group; the MOCK group only dealed with Lipofectamine 2000. Morphological changes of RD cells in each group were observed under the microscopy, viral RNA and protein were tested by quantitative RTPCR and western blotting, respectively. Results: The maximal transfection efficiency occurred when the siRNA concentration was at 60 nmol/L, the transfection efficiency was up to 87%. RD cells of siRNA2Apro group showed slight cytopathic effect and a higher cell survival rate compared to control groups, through the observation of the morphological changes of the cells infected with EV71. The results of quantitative RTPCR and western blotting revealed that the levels of viral RNA and protein of siRNA2Apro group were significantly lower than the control group (P<0.01). Conclusion: siRNAs targeted the 2Apro genomic region of EV71 exerts potential antiviral effects.
收稿日期: 2014-04-07
基金资助:国家自然科学基金资助项目（81370084）；镇江市社会发展项目（SH2012036）
通讯作者:
陈建国（通讯作者），主任技师，E-mail:cjg02@126.com
作者简介: 刘海冰（1987—），男，硕士研究生；
引用本文:
刘海冰1， 孔枕枕2， 赵培培2， 茅凌翔3， 苏兆亮2， 陈建国1. 干扰EV71病毒2Apro基因的siRNA具有潜在的抗病毒作用[J]. 江苏大学学报:医学版, 2014, 24(05): 403-407.
LIU Hai-bing1, KONG Zhen-zhen2, ZHAO Pei-pei2, MAO Ling-xiang3, SU Zhao-liang2, CHEN Jian-guo1. siRNA against the 2Apro genomic region of EV71 exerts potential antiviral effects. Journal of Jiangsu University(Medicine Edition), 2014, 24(05): 403-407.
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2014, Vol. 24 
