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

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.