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MiR-490对B组3型柯萨奇病毒复制的抑制作用

The Suppressive Effect of MiR-490 on Coxsackievirus B3 Replication
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摘要 为了研究miR-490对柯萨奇病毒B组3型(Coxsackievirus B type 3,CVB3)的作用,本文采用体外实验将miR-490转染入HeLa细胞,用带有海肾荧光素酶(Renilla luciferase,RLuc)基因的CVB3重组体(RLuc-CVB3)感染HeLa细胞,感染后0~40h,每隔8h收获一次细胞,测定荧光素酶活性,观察miR-490对RLuc-CVB3复制的作用。同时在另一实验组我们用带有绿色荧光(Enhanced green fluorescence protein,EGFP)表达的CVB3重组体(EGFP-CVB3)攻击表达miR-490的HeLa细胞和用反义miR-490(AMO-miR-490)沉默miR-490表达的HeLa细胞,观察miR-490对EGFP-CVB3复制和表达的影响。结果我们发现miR-490抑制RLuc-CVB3复制,miR-490在RLuc-CVB3感染后32h抑制效果最显著(P〈0.05)。同时发现在感染EGFP-CVB3后32h,miR-490对EGFPCVB3的复制和表达也有显著的抑制作用,但抑制作用可以被反义miR-490完全抵消。因此我们得出结论,miR-490能够显著抑制CVB3的复制和表达。 To study the effect of miR-490 on Coxsackievirus B3(CVB3)replication,HeLa cells were transfected with miR-490 in vitro,and infected with a Renilla luciferase(RLuc)-expressing CVB3variant(RLuc-CVB3).The activities of RLuc in these cells were measured at 8hintervals from 0to 40 hpost.-infection(p.i.),and the effects of miR-490 on RLuc-CVB3 replication were observed.In a further study,HeLa cells were transfected with either miR-490 or antisense miR-490(AMO-miR-490),and were then infected with an enhanced green fluorescence protein(EGFP)-expressing CVB3variant(EGFP-CVB3).The replication of EGFP-CVB3 was then determined by detecting the expression of EGFP.We observed that miR-490 could significantly inhibit the expression of RLuc in infected cells at 32 hp.i.Furthermore,in HeLa cells infected with EGFP-CVB3 at 32hp.i.,EGFP expression was also significantly inhibited by the presence of miR-490.The inhibitory effect of miR-490 on EGFP expression in EGFP-CVB3-infected cells could be reversed by transfection with AMO-miR-490.These results indicated that miR-490 significantly inhibits the replication and expression of CVB3.
出处 《病毒学报》 CAS CSCD 北大核心 2014年第6期619-623,共5页 Chinese Journal of Virology
基金 黑龙江省卫生厅资助项目(2012-535)
关键词 柯萨奇病毒 miR-490 抑制 病毒复制 Coxsackievirus MiR-490 Suppression Virus replication
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参考文献13

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