College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi China.
Medical College of QingHai University, Xining, Qinghai, China.
Virulence. 2020 Dec;11(1):260-269. doi: 10.1080/21505594.2020.1735051.
Classical swine fever virus (CSFV) is one of the most important viral pathogens leading worldwide threats to pig industry. MicroRNAs (miRNAs) play important roles in regulating virus replication, but whether miRNAs affect CSFV infection is still poorly understood. In previous study, we identified four miRNAs that were down-regulated by CSFV in swine umbilical vein endothelial cells (SUVEC). In this study, miR-140, one of the most potently down-regulated genes was investigated. We found that the miRNA expression was significantly inhibited by CSFV infection. Subsequent studies revealed that miR-140 mimics significantly inhibited CSFV replication, while the inhibition of endogenous miR-140 enhanced CSFV replication. By using bioinformatics prediction, luciferase reporter system, real-time fluorescence quantitative PCR (RT-qPCR) and Western blot assays, we further demonstrated that miR-140 bind to the 3' UTR of Rab25 mRNA to regulate its expression. We also analyzed the expression pattern of Rab25 in SUVECs after CSFV infection. The results showed that CSFV infection induced Rab25 expression. Finally, Rab25 was found to promote CSFV replication. In conclusion, this study demonstrated that CSFV inhibits miR-140 expression and miR-140 inhibits replication by binding to host factor Rab25.
经典猪瘟病毒(CSFV)是一种重要的病毒病原体,对全球养猪业构成严重威胁。microRNAs(miRNAs)在调节病毒复制中发挥着重要作用,但miRNAs 是否影响 CSFV 感染尚不清楚。在之前的研究中,我们鉴定了 CSFV 在猪脐静脉内皮细胞(SUVEC)中下调的 4 个 miRNAs。在本研究中,我们研究了下调最明显的一个 miRNA:miR-140。我们发现 CSFV 感染显著抑制了 miRNA 的表达。进一步的研究表明,miR-140 模拟物显著抑制了 CSFV 的复制,而内源性 miR-140 的抑制增强了 CSFV 的复制。通过生物信息学预测、荧光素酶报告系统、实时荧光定量 PCR(RT-qPCR)和 Western blot 分析,我们进一步证实了 miR-140 通过结合 Rab25 mRNA 的 3'UTR 来调节其表达。我们还分析了 CSFV 感染后 SUVEC 中 Rab25 的表达模式。结果表明,CSFV 感染诱导了 Rab25 的表达。最后,发现 Rab25 促进了 CSFV 的复制。总之,本研究表明 CSFV 通过抑制 miR-140 的表达,并通过结合宿主因子 Rab25 来抑制其复制。