miR-545 promoted enterovirus 71 replication via directly targeting phosphatase and tensin homolog and tumor necrosis factor receptor-associated factor 6
miR-545 promoted enterovirus 71 replication via directly targeting phosphatase and tensin homolog and tumor necrosis factor receptor-associated factor 6
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DOI：
10.1002/jcp.28222
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发表时间：
2019-09-01
影响因子：
5.6
通讯作者：
Zhang, Xiaoli
中科院分区：
文献类型：
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作者：
Sun, Ying;Feng, Long;Zhang, Xiaoli
肠道病毒71型（Enterovirus 71，EV 71）是一种无包膜的二十面体RNA病毒，是引起手足口病的主要病原体。近年来，microRNAs（miRNAs）在EV 71病毒复制的发病机制中发挥重要作用。本研究旨在探讨miR-545在EV 71病毒复制中的作用及其分子机制。我们发现miR-545在EV 71感染的人胚肾（HEK）293细胞和横纹肌肉瘤（RD）细胞中上调。在HEK 293和RD细胞中，过表达miR-545可促进EV 71的病毒复制，并减弱EV 71对细胞活力的抑制作用;而敲低miR-545则可显著抑制EV 71在这两种细胞中的复制。生物信息学分析和荧光素酶报告基因检测结果表明，miR-545可直接靶向HEK 293细胞中磷酸酶和张力蛋白同源物（PTEN）和肿瘤坏死因子受体相关因子6（TRAF 6）的3个非翻译区。此外，miR-545负调节PTEN和TRAF 6的信使RNA（mRNA）和蛋白表达。在HEK 293细胞中，EV 71感染也抑制了PTEN和TRAF 6的mRNA和蛋白表达，这通过miR-545敲低而减弱。过表达PTEN和TRAF 6均能抑制HKE 293细胞中EV 71的复制，并减弱过表达miR-545对HEK 293细胞中EV 71复制的增强作用。总的来说，我们的研究首次表明miR-545对EV 71在HEK 293和RD细胞中的复制具有增强作用。进一步的机制结果表明，miR-545至少部分通过靶向PTEN和TRAF 6促进EV 71复制。
Enterovirus 71 (EV71) is a small, nonenveloped icosahedral RNA virus and is the predominant causative pathogen of hand-foot-and-mouth disease. Recently, microRNAs (miRNAs) are reported to play important roles in the pathogenesis of EV71 replication. This study investigated the role of miR-545 in the EV71 replication and explored the underlying molecular mechanisms. We showed that miR-545 was upregulated in the EV71-infected human embryonic kidney (HEK) 293 cells and rhabdomyosarcoma (RD) cells. Overexpression of miR-545 promoted the viral replication of EV71 and attenuated the inhibitory effects of EV71 on cell viability in HEK293 and RD cells; while knockdown of miR-545 significantly suppressed the EV71 replication in these two cell lines. Bioinformatics analysis and luciferase reporter assay showed that miR-545 directly targeted the 3untranslated region of phosphatase and tensin homolog (PTEN) and tumor necrosis factor receptor-associated factor 6 (TRAF6) in HEK293 cells. Furthermore, miR-545 negatively regulated the messenger RNA (mRNA) and protein expression of PTEN and TRAF6. The mRNA and protein expression of PTEN and TRAF6 was also suppressed by EV71 infection, which was attenuated by miR-545 knockdown in HEK293 cells. Overexpression of PTEN and TRAF6 both suppressed the EV71 replication in HKE293 cells, and also attenuated the enhanced effects of miR-545 overexpression on the EV71 replication in HEK293 cells. Collectively, our study for the first time showed that miR-545 had an enhanced effect on the EV71 replication in HEK293 and RD cells. Further mechanistic results indicated that miR-545 promoted EV71 replication at least partly via targeting PTEN and TRAF6.