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微小RNA-27b通过靶向 runt 相关转录因子1(RUNX1)减轻传染性胃肠炎病毒(TGEV)感染诱导的细胞凋亡。

miR-27b attenuates apoptosis induced by transmissible gastroenteritis virus (TGEV) infection via targeting runt-related transcription factor 1 (RUNX1).

作者信息

Zhao Xiaomin, Song Xiangjun, Bai Xiaoyuan, Fei Naijiao, Huang Yong, Zhao Zhimin, Du Qian, Zhang Hongling, Zhang Liang, Tong Dewen

机构信息

College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, China.

出版信息

PeerJ. 2016 Feb 4;4:e1635. doi: 10.7717/peerj.1635. eCollection 2016.

Abstract

Transmissible gastroenteritis virus (TGEV), belonging to the coronaviridae family, is the key cause of the fatal diarrhea of piglets and results in many pathological processes. microRNAs (miRNAs) play a key role in the regulation of virus-induced apoptosis. During the process of apoptosis induced by TGEV infection in PK-15 cells, the miR-27b is notably down-regulated. Thus, we speculate that miR-27b is involved in regulating the process of apoptosis in PK-15 cells. In this study we demonstrated that the over-expression of miR-27b led to the inhibition of TGEV-induced apoptosis, reduction of Bax protein level, and decrease of caspase-3 and -9 activities. Conversely, silencing of miR-27b by miR-27b inhibitors enhanced apoptosis via up-regulating Bax expression and promoting the activities of caspase-3 and -9 in TGEV-infected cells. Subsequently, the runt-related transcription factor 1 (RUNX1) is a candidate target of miR-27b predicted by bioinformatics search. We further identified that the miR-27b directly bound to the 3' UTR of RUNX1 mRNA and suppressed RUNX1 expression, which indicates RUNX1 is the direct target gene of miR-27b. The over-expression of RUNX1 increased apoptosis and knockdown RUNX1blocked apoptosis of viral-infected cells via regulating Bax expression and the activities of caspase-3 and -9. Our data reveal that miR-27b may repress the mitochondrial pathway of apoptosis by targeting RUNX1, indicating that TGEV may induce apoptosis via down-regulating miR-27b and that miR-27b may act as a target for therapeutic intervention.

摘要

传染性胃肠炎病毒(TGEV)属于冠状病毒科,是仔猪致命性腹泻的主要病因,并引发多种病理过程。微小RNA(miRNA)在病毒诱导的细胞凋亡调控中起关键作用。在TGEV感染PK - 15细胞诱导凋亡的过程中,miR - 27b显著下调。因此,我们推测miR - 27b参与调控PK - 15细胞的凋亡过程。在本研究中,我们证明miR - 27b的过表达导致TGEV诱导的凋亡受到抑制,Bax蛋白水平降低,以及caspase - 3和 - 9活性下降。相反,用miR - 27b抑制剂沉默miR - 27b可通过上调Bax表达并促进TGEV感染细胞中caspase - 3和 - 9的活性来增强细胞凋亡。随后,通过生物信息学搜索预测,与 runt 相关的转录因子1(RUNX1)是miR - 27b的候选靶标。我们进一步鉴定出miR - 27b直接与RUNX1 mRNA的3'UTR结合并抑制RUNX1表达,这表明RUNX1是miR - 27b的直接靶基因。RUNX1的过表达增加细胞凋亡,而敲低RUNX1可通过调节Bax表达以及caspase - 3和 - 9的活性来阻止病毒感染细胞的凋亡。我们的数据表明,miR - 27b可能通过靶向RUNX1抑制细胞凋亡的线粒体途径,这表明TGEV可能通过下调miR - 27b诱导细胞凋亡,并且miR - 27b可能成为治疗干预的靶点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/df26/4748701/e510a6efc3d2/peerj-04-1635-g001.jpg

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