宿主蛋白LSml-7复合体在登革病毒RNA复制中的作用研究

目的探讨LSml-7复合体在登革病毒（DENV）RNA复制中的作用。方法DENV感染HepG2细胞后．在不同时间点（2、24、36、48h）收集细胞和提取总mRNA，实时定量PCR（RT-qPCR）分析LSml表达水平；将LSml的siRNA和非特异对照siRNA两次转染HepG2细胞后感染DENV-2，24h后收集细胞，提取总mRNA，RT-qPCR分析LSml表达水平与病毒RNA表达水平；利用LSml抗体与dsRNA抗体对DENV感染细胞进行免疫荧光染色，激光共聚焦分析LSml与dsRNA共定位情况。结果与2h比较，随着时间的推移，DENV-2RNA和LSmlRNA水平逐渐升高；与对照组siNC比较，siLSml组LSmlRNA水平明显降低，沉默效率达78．2％；与对照组siNC比较，siLSml组DENVRNA含量降低35．6％：LSml-7复合体在DENV感染HepG2细胞后募集到核周围，与DENVdsRNA共定位。结论LSml-7复合体可能作为DENV复制复合物的组成部分，正调控DENVRNA的复制。

The role of host factor LSml-7 complex in dengue virus RNA replication

Objective To investigate the roles of host protein LSml-7 complex in DENV RNA replication. Methods mRNA was isolated from DENV infected HepG2 cells at different time point（2,24,36,48 h） of post-infection, and the level of DENV RNA and LSml RNA was analysed by RT-qPCR. HepG2 cells were transfected with specific siRNA for LSml and nontargeting siRNA and siNC respectively,followed by infection with DENV.The mRNA was isolated at 24 h p. i. and analyzed by RT-qPCR to determine the RNA level of DENV and LSml. Laser scanning eonfoeal microscopy was performed to observe the localization of LSml protein and DENV dsRNA in DENV infected HepG2 cells. Results The levels of DENV RNA and LSml RNA were increased compared to 2 h p.i. during the course of DENV infection, siRNA- mediated silencing resulted in a specific 78.2% reduction of LSml RNA when using the nontargeting siNC as a negative control, and down-regulation of LSml resulted in a marked reduction of the DENY RNA level by 35.6%. LSml-7 complex distributed and colocalized with DENV dsRNA Conclusion LSml-7 complex may act as the component regulators for DENV RNA replication. around nucleolous in the DENV infected HepG2 cells. of DENV replication machinery and is one of the positive