The role of miR-320a and IL-1β in human chondrocyte degradation

Abstract

ObjectivesThis study aimed to explore the role of miR-320a in the pathogenesis of osteoarthritis (OA).MethodsHuman cartilage cells (C28/I2) were transfected with miR-320a or antisense oligonucleotides (ASO)-miR-320a, and treated with IL-1β. Subsequently the expression of collagen type II alpha 1 (Col2α1) and aggrecan (ACAN), and the concentrations of sulfated glycosaminoglycans (sGAG) and matrix metallopeptidase 13 (MMP-13), were assessed. Luciferase reporter assay, qRT-PCR, and Western blot were performed to explore whether pre-B-cell leukemia Homeobox 3 (PBX3) was a target of miR-320a. Furthermore, cells were co-transfected with miR-320a and PBX3 expressing vector, or cells were transfected with miR-320a and treated with a nuclear factor kappa B (NF-κB) antagonist MG132. The changes in Col2α1 and ACAN expression, and in sGAG and MMP-13 concentrations, were measured again. Statistical comparisons were made between two groups by using the two-tailed paired t-test.ResultsExpression of miR-320a was elevated in OA cartilage tissues and chondrocytes, and in IL-1β-stimulated C28/I2 cells (p < 0.05 or p < 0.01). MiR-320a overexpression enhanced IL-1β-induced down-regulation of Col2α1 and ACAN and sGAG, and increased the IL-1β-induced overexpression of MMP-13 (p < 0.01). PBX3 was a direct target of miR-320a. PBX3 and MG132 co-transfection attenuated the effects of miR-320a on the expression of Col2α1, ACAN, sGAG and MMP-13(p < 0.01).ConclusionOverexpression of miR-320a might enhance IL-1β-induced cartilage degradation factors. These effects might be via targeting PBX3 and regulating NF-κB.Cite this article: Y. Jin, X. Chen, Z. Y. Gao, K. Liu, Y. Hou, J. Zheng. The role of miR-320a and IL-1β in human chondrocyte degradation. Bone Joint Res 2017;6:–203. DOI: 10.1302/2046-3758.64.BJR-2016-0224.R1.

MicroRNAs and Osteoarthritis

“…A few years ago, Meng et al [ 82 ] also identified miR-320 as a regulator of MMP13 , where NF-κB and MAPK activation was shown to downregulate miR-320 expression. This conclusion was later extended by Jin et al [ 83 ], who showed that overexpression of miR-320a, which targets the pre-B-cell leukemia transcription factor 3 ( PBX3 ) gene, increased MMP-13, whereas PBX3 and the proteasome inhibitor, MG132, suppressed the effects of miR-320a on the expression of COL2A1 , ACAN (i.e., the aggrecan gene), sulfated glycosaminoglycans, and MMP-13. Moreover, miR-105 was also found to be important for the fibroblast growth factor-2 (FGF2)-induced gene expression of ADAMTS4 , 5 , 7 , and 12 [ 84 ].…”

Origin and Efficacy of Hyaluronan Injections in Knee Osteoarthritis: Randomized, Double-Blind Trial

“…The Wilcoxon test following Bonferroni adjustment was performed for VAS analog, WOMAC score, Lequesne Index score, TUG test, and SLS test between baseline and after 1 month, 3 months, and 6 months follow-up [ 21 ]. Independent t -test [ 22 ] following Bonferroni adjustment [ 21 ] was performed for VAS analog, WOMAC score, Lequesne Index score, patient satisfaction, and adverse effects between groups at 1-month, 3-months, and 6-months follow-up. The Mann-Whitney U test following Bonferroni adjustment was performed for the TUG test and the SLS test between the groups at 6-month follow-up [ 21 ].…”

Pro-osteoporotic miR-320a impairs osteoblast function and induces oxidative stress

“…For example, a microarray analysis has shown overexpression of miR-320a in whole bone tissue from women with osteoporotic fracture [2, 3]. Moreover, miR-320a is elevated in osteoarthritic cartilage tissue and chondrocytes [4], highlighting its potential functionality in the musculoskeletal system. This miRNA has been reported to have multiple mRNA targets and to regulate different pathways in various cell types.…”