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miR-927 在 C6/36 蚊细胞中感染登革热病毒 2 型的急性和持续性感染中具有促病毒作用。

miR-927 has pro-viral effects during acute and persistent infection with dengue virus type 2 in C6/36 mosquito cells.

机构信息

Programa de Doctorado en Ciencias en Biotecnología, Escuela Nacional de Medicina y Homeopatía, Instituto Politécnico Nacional, Guillermo Massieu Helguera 249, La Escalera-Ticomán, Mexico City CP 07320, Mexico.

Posgrado en Ciencias Genómicas, Universidad Autónoma de la Ciudad de México, San Lorenzo 290, Del Valle, Mexico City CP 03100, Mexico.

出版信息

J Gen Virol. 2020 Aug;101(8):825-839. doi: 10.1099/jgv.0.001441. Epub 2020 Jun 1.

Abstract

Dengue virus (DENV) is an important flavivirus that is transmitted to humans by mosquitoes, where it can establish a persistent infection underlying vertical and horizontal transmission. However, the exact mechanism of persistent DENV infection is not well understood. Recently miR-927 was found to be upregulated in C6/36-HT cells at 57 weeks of persistent infection (C6-L57), suggesting its participation during this type of infection. The aim of this study was to determine the role of miR-927 during infection with DENV type 2. The results indicate an overexpression of miR-927 in C6-L57 cells and acutely infected cells according to the time of infection and the m.o.i. used. The downregulation of miR-927 in C6-L57 cells results in a reduction of both viral titre and viral genome copy number. The overexpression of miR-927 in C6-L40 and C6/36 cells infected at an m.o.i. of 0.1 causes an increase in both viral titre and viral genome copy number, suggesting a pro-viral activity of miR-927. prediction analysis reveals target mRNAs for miR-927 are implicated in post-translational modifications (SUMO), translation factors (eIF-2B), the innate immune system (NKIRAS), exocytosis (EXOC-2), endocytosis (APM1) and the cytoskeleton (FLN). The expression levels of FLN were the most affected by both miR-927 overexpression and inhibition, and FLN was determined to be a direct target of miR-927 by a dual-luciferase gene reporter assay. FLN has been associated with the regulation of the Toll pathway and either overexpression or downregulation of miR-927 resulted in expression changes of antimicrobial peptides (Cecropins A and G, and Defensin D) involved in the Toll pathway response.

摘要

登革病毒(DENV)是一种重要的黄病毒,通过蚊子传播给人类,在蚊子体内可建立垂直和水平传播的持续感染。然而,持续 DENV 感染的确切机制尚不清楚。最近发现 miR-927 在持续感染 57 周的 C6/36-HT 细胞(C6-L57)中上调,提示其参与了这种类型的感染。本研究旨在确定 miR-927 在感染登革热病毒 2 型时的作用。结果表明,根据感染时间和感染复数(m.o.i.),miR-927 在 C6-L57 细胞和急性感染细胞中呈过度表达。C6-L57 细胞中 miR-927 的下调导致病毒滴度和病毒基因组拷贝数均降低。在感染复数(m.o.i.)为 0.1 的 C6-L40 和 C6/36 细胞中过表达 miR-927 会导致病毒滴度和病毒基因组拷贝数均增加,表明 miR-927 具有促病毒活性。预测分析显示,miR-927 的靶 mRNA 参与翻译后修饰(SUMO)、翻译因子(eIF-2B)、先天免疫系统(NKIRAS)、胞吐作用(EXOC-2)、内吞作用(APM1)和细胞骨架(FLN)。FLN 的表达水平受 miR-927 的过表达和抑制的影响最大,通过双荧光素酶基因报告基因检测确定 FLN 是 miR-927 的直接靶标。FLN 与 Toll 途径的调节有关,miR-927 的过表达或下调均导致参与 Toll 途径反应的抗菌肽(Cecropins A 和 G 和 Defensin D)的表达发生变化。

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