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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。
In order to investigate the effect of miR-490 on Coxsackievirus B type 3 (CVB3), miR-490 was transfected into HeLa cells in vitro and transfected with Renilla luciferase (RLuc-CVB3) were transfected into HeLa cells. The cells were harvested at 0 ~ 40 h after infection and the luciferase activity was measured at 8 h after transfection. The effect of miR-490 on RLuc-CVB3 replication was observed. At the same time, in another experimental group, we treated HeLa cells expressing miR-490 with CVB3 recombinant (EGFP-CVB3) expressing with enhanced green fluorescence protein (EGFP) and antisense miR-490 490) silencing miR-490-expressing HeLa cells and observing the effect of miR-490 on EGFP-CVB3 replication and expression. Results We found that miR-490 inhibited RLuc-CVB3 replication, and miR-490 had the most significant inhibitory effect at 32h after RLuc-CVB3 infection (P <0.05). At the same time, it was found that miR-490 had a significant inhibitory effect on EGFPCVB3 replication and expression at 32h after infection with EGFP-CVB3, but the inhibitory effect could be completely counteracted by antisense miR-490. Therefore, we conclude that miR-490 significantly inhibits the replication and expression of CVB3.