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Vero细胞中miR - 484和miR - 744的过表达改变登革病毒复制。

Overexpression of miR-484 and miR-744 in Vero cells alters Dengue virus replication.

作者信息

Castrillón-Betancur Juan Camilo, Urcuqui-Inchima Silvio

机构信息

Universidad de Antioquia, Facultad de Medicina, Grupo Inmunovirología, Medellín, Colombia.

出版信息

Mem Inst Oswaldo Cruz. 2017 Apr;112(4):281-291. doi: 10.1590/0074-02760160404. Epub 2017 Mar 2.

Abstract

BACKGROUND

Dengue is considered one of the world's most important mosquito-borne diseases. MicroRNAs (miRNAs) are small non-coding single-stranded RNAs that play an important role in the regulation of gene expression in eukaryotes. Although miRNAs possess antiviral activity against many mammalian-infecting viruses, their involvement in Dengue virus (DENV) replication remains poorly understood.

OBJECTIVE

To determine the role of miR-484 and miR-744 in DENV infection and to examine whether DENV infection alters the expression of both miRNAs.

METHODS

We used bioinformatics tools to explore the relationship between DENV and cellular miRNAs. We then overexpressed miR-484 or miR-744 in Vero cells to examine their role in DENV replication using flow cytometry, reverse transcriptase quantitative polymerase chain reaction (RT-qPCR), and western blotting.

FINDINGS

We found several cellular miRNAs that target a conserved region within the 3' untranslated region (3' UTR) of the genome of the four DENV serotypes and found that overexpression of miR-484 or miR-744 inhibits infection by DENV-1 to DENV-4. Furthermore, we observed that DENV RNA might be involved in the downregulation of endogenous miR-484 and miR-744.

CONCLUSION

Our study identifies miR-484 and miR-744 as two possible restriction host factors against DENV infection. However, further studies are needed to directly verify whether miR-484 and miR-744 both have an anti-DENV effect in vivo.

摘要

背景

登革热被认为是世界上最重要的蚊媒疾病之一。微小RNA(miRNA)是小的非编码单链RNA,在真核生物基因表达调控中发挥重要作用。尽管miRNA对许多感染哺乳动物的病毒具有抗病毒活性,但其在登革热病毒(DENV)复制中的作用仍知之甚少。

目的

确定miR-484和miR-744在DENV感染中的作用,并检测DENV感染是否会改变这两种miRNA的表达。

方法

我们使用生物信息学工具探索DENV与细胞miRNA之间的关系。然后在Vero细胞中过表达miR-484或miR-744,使用流式细胞术、逆转录定量聚合酶链反应(RT-qPCR)和蛋白质印迹法检测它们在DENV复制中的作用。

结果

我们发现了几种靶向四种DENV血清型基因组3'非翻译区(3'UTR)内保守区域的细胞miRNA,并发现过表达miR-484或miR-744可抑制DENV-1至DENV-4的感染。此外,我们观察到DENV RNA可能参与内源性miR-484和miR-744的下调。

结论

我们的研究确定miR-484和miR-744是两种可能的抗DENV感染的宿主限制因子。然而,需要进一步研究直接验证miR-484和miR-744在体内是否均具有抗DENV作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e045/5354610/7ffec0b27128/0074-0276-mioc-0074-02760160404-gf01.jpg

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