Analysis of the miRNA profile in C6/36 cells persistently infected with dengue virus type 2
Analysis of the miRNA profile in C6/36 cells persistently infected with dengue virus type 2
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DOI：
10.1016/j.virusres.2017.03.005
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发表时间：
2017-03-15
期刊：
影响因子：
5
通讯作者：
Santiago Salas-Benito, Juan
中科院分区：
文献类型：
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作者：
Gamaliel Avila-Bonilla, Rodolfo;Yocupicio-Monroy, Martha;Santiago Salas-Benito, Juan
登革病毒（DENV）是世界上最重要的虫媒病毒; DENV由伊蚊属的蚊子传播，并且可以在蚊子中建立终身持续感染。然而，建立持续感染的确切机制仍然未知。在这项研究中，使用先前建立的持续感染DENV 2（C6-L）的C6/36-HT细胞系作为模型，通过深度测序和RT-qPCR分析miRNA的差异表达。miR-927、miR-87、miR-210、miR-2a-3p、miR-190和miR-970上调，而miR-252、miR-263a-3p、miR-92b、miR-10-5p、miR-9a-5p、miR-9a-1、miR-124、与用相同病毒急性感染的C6/36细胞或模拟感染的细胞相比，miR-286 a和miR-286 b在C6-L细胞中下调。通过RT-qPCR验证了高度差异表达的miR-927和miR-9a-5 p的深度测序结果，与急性和模拟感染的C6/36细胞相比，它们分别上调和下调。这些miRNA的推定靶点包括泛素结合途径、囊泡介导的转运、自噬和JAK-STAT级联以及具有内肽酶活性的蛋白质的组分。其他推定的靶标包括Toll信号传导途径的成员和具有激酶、ATP酶、蛋白酶、清道夫受体或凝集素C型活性或参与脂肪酸生物合成或氧化应激的蛋白质。我们的研究结果表明，几种特定的miRNA有助于调节细胞功能，在蚊子细胞的持续感染期间维持病毒复制和抗病毒反应之间的平衡。这项研究是第一次报道在持续感染DENV的蚊子细胞系中的全球miRNA谱。(C)2017爱思唯尔B. V.保留所有权利。
Dengue virus (DENV) is the most important arbovirus in the world; DENV is transmitted by the Aedes genus of mosquitoes and can establish a life-long persistent infection in mosquitoes. However, the exact mechanism by which persistent infection is established remains unknown. In this study the differential expression of miRNAs was analysed by deep sequencing and RT-qPCR using a previously established C6/36-HT cell line persistently infected with DENV 2 (C6-L) as a model. miR-927, miR-87, miR-210, miR-2a-3p, miR-190 and miR-970 were up-regulated, whereas miR-252, miR-263a-3p, miR-92b, miR-10-5p miR-9a-5p, miR-9a-1, miR-124, miR-286a and miR-286b were down-regulated in C6-L cells compared with C6/36 cells acutely infected with the same virus or mock-infected cells. Deep sequencing results were validated by RT-qPCR for the highly differentially expressed miR-927 and miR-9a-5p, which were up- and down-regulated, respectively, compared with both acutely and mock-infected C6/36 cells. The putative targets of these miRNAs include components of the ubiquitin conjugation pathway, vesicle mediated transport, autophagy, and the JAK-STAT cascade as well as proteins with endopeptidase activity. Other putative targets include members of the Toll signalling pathway and proteins with kinase, ATPase, protease, scavenger receptor or Lectin C-type activity or that participate in fatty acid biosynthesis or oxidative stress. Our results suggest that several specific miRNAs help regulate the cellular functions that maintain equilibrium between viral replication and the antiviral response during persistent infection of mosquito cells. This study is the first report of a global miRNA profile in a mosquito cell line persistently infected with DENV. (C) 2017 Elsevier B.V. All rights reserved.