microRNA-124-3p靶向调控STAT3抑制神经细胞中肠道病毒71型的复制

microRNA-124-3p inhibits enterovirus 71 replication in nerve cells via targeting STAT3

目的构建microRNA-124-3p表达上调和下调的SK-N-SH细胞,探讨microRNA-124-3p对肠道病毒71型(EV71)在SK-N-SH中的复制机制。方法构建过表达miRNA的miRNA mimic和敲低miRNA的miRNA inhibitor。检测miRNA-124-3p,EV71核酸,细胞病变效应(CPE),病毒滴度TCID_(50),Ratio值,VP1、STAT3、P-STAT3、CC-ND2、MMP2蛋白表达。结果与miR-124-3p mimic NC组比,miR-124-3p mimic组miR-124-3p上升,LgTCID_(50)降低,EV71核酸的复制受抑,转染野生型miRSTAT3-WT质粒细胞活性被上调的miRNA-124-3p抑制,STAT3、P-STAT3、CC-ND2和MMP2降低(均P<0.05),与miR-124-3p inhibitorNC组比,miR-124-3p inhibitor组miR-124-3p降低,CPE增高,STAT3、P-STAT3、CC-ND2和MMP2升高(均P<0.05)。结论EV71感染SK-N-SH神经细胞,复制受抑,与microRNA-124-3p靶向抑制STAT通路分子STAT3、P-STAT3、CCND2和MMP2有关。

To study the effect of microRNA-124-3 p on enterovirus 71(EV71) in SK-N-SH nerve cells, SKN-SH nerve cells with up-or down-regulated expression of microRNA-124-3 p were constructed, and the regulation and potential mechanism of action in the replication process was analyzed. The nerve cells were divided into miR-124-3 p inhibitor group, miR-124-3 p inhibitor NC group, miR-124-3 p-mimic group and miR-124-3 p mimic NC group. And then we detected the relative expression of miRNA-124-3 p, EV71 nucleic acid, cytopathic effect(CPE), TCID_(50), ratio, VP1, STAT3, P-STAT3, CC-ND2 and MMP2 of each group. Data showed that as compared with miR-124-3 p mimic NC, miR-124-3 p mimic group demonstrated higher miR-124-3 p level, lower LgTCID_(50), inhibited EV71 nucleic acid replication, inhibited cell activity, and lower levels of STAT3, P-STAT3,CC-ND2 and MMP2(all P<0.05);compared with miR-124-3 p inhibitor NC, miR-124-3 p inhibitor group showed lower miR-124-3 p, higher levels of CPE, STAT3, P-STAT3, CC-ND2 and MMP2(all P<0.05). In conclusion, the replication of EV71 is inhibited in SK-N-SH nerve cells, and this inhibitory effect is achieved by microRNA-124-3 p inhibiting the protein expression levels of STAT pathway molecules STAT3, P-STAT3, CCND2 and MMP2.