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Up-regulated miR-374a-3p relieves lipopolysaccharides induced injury in CHON-001 cells via regulating Wingless-type MMTV integration site family member 5B.

Abstract：

BACKGROUND:Osteoarthritis (OA) is a frequent and incurable joint disease, inducing significant pain and seriously threatening to human health. It has been reported that microRNAs (miRNAs) play crucial roles on cancers and inflammatory diseases via cooperating with genes. However, the effect of miR-374a-3p/Wingless-type MMTV integration site family, member 5B (WNT5B) pair in OA remains to be explored. METHODS:GSE105027 and GSE55457 datasets were obtained to reveal the expression of miR-374a-3p and WNT5B in OA cartilages using log-scale. The OA cell model was established by lipopolysaccharides (LPS) stimulation in CHON-001 cells and the functional role of miR-374a-3p on OA was investigated by analyzing cell proliferation, cell apoptosis and the expression of apoptosis-related proteins (Bcl-2, Bax and Bim). Through bioinformatics prediction, WNT5B, the target gene of miR-374a-3p, was predicted and the association between miR-374a-3p and WNT5B was further explored by luciferase reporter assay. Functional experiments in vitro were conducted to assess whether WNT5B was involved in the regulation of miR-374a-3p to LPS-stimulated CHON-001. Finally, the expression of JNK/ERK/MAPK pathway-related proteins was detected to explore the underlying molecular mechanism. RESULTS:The data set showed that miR-374a-3p was decreased in OA cartilages and the consistent expressional pattern was observed in LPS-stimulated CHON-001 cells. Overexpression of miR-374a-3p significantly alleviated LPS-induced damage in CHON-001 cells, whereas miR-374a-3p inhibitor aggravated LPS-stimulated injury. Further experiments demonstrated that WNT5B was a target of miR-374a-3p and its expression was decreased by miR-374a-3p. WNT5B expression was increased in OA cartilages. Silencing WNT5B prevented CHON-001 cells from LPS-induced damage. Down-regulation of WNT5B strengthened the protective effect of miR-374a-3p on LPS-stimulated CHON-001 cells. Moreover, miR-374a-3p cooperated with WNT5B to affect cell behaviors of LPS-stimulated CHON-001 cells via mediating the JNK/ERK/MAPK pathway. CONCLUSION:These results indicated that overexpression of miR-374a-3p protects CHON-001 cells against LPS challenge by modulating WNT5B and inhibiting the JNK/ERK/MAPK pathway.

journal_name

Mol Cell Probes

journal_title

Molecular and cellular probes

authors

Shi FL,Ren LX

doi

10.1016/j.mcp.2020.101541

subject

Has Abstract

pub_date

2020-06-01 00:00:00

pages

101541

eissn

0890-8508

issn

1096-1194

pii

S0890-8508(19)30492-X

journal_volume

pub_type

杂志文章

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Up-regulated miR-374a-3p relieves lipopolysaccharides induced injury in CHON-001 cells via regulating Wingless-type MMTV integration site family member 5B.