MiR-143 Inhibits Osteoclastogenesis by Targeting RANK and NF-κB and MAPK Signaling Pathways.
OBJECTIVE
To explore the effect of miRNA-143 on osteoclast formation and provide new ideas for the treatment of osteoporosis.


METHODS
Mice macrophage lines RAW264.7 cells after transfection were divided into four groups: control group, RANKL group, RANKL combined with miR-143 mimics group and RANKL combined with miR-NC group. TARCP staining was used to observe the effect of miR-143 on osteoclast formation. The expression of RANK, TRAF6 and NFATc-1 in the upstream of RANKL pathway was detected by real-time quantitative PCR (RT qPCR) and Western blotting (WB). The binding of miR-143 to TNFRSF11A was detected by double Luciferase Reporter Analysis. The effect of miR-143 on the expression of NF-κB (p65, I-κB-α) signal pathway in osteoclasts was detected. The effects of I-BET151 on the expression of osteoclast-specific genes TRACP, MMP 9, CtsK and c-Src were detected.


RESULTS
The positive level of osteoclasts in RANKL group and RANKL combined with miR-NC group was significantly higher than that of RANKL combined with miR-143 mimics group and control group (P < 0.05). The expression levels of RANK, TRAF6, NFATc-1, TRACP, MMP-9, CtsK and c-Src in RANKL group and RANKL combined with miR-NC group were significantly higher than those of RANKL combined with miR-143 mimics group and control group (P < 0.05). The expression levels of I-κB-α were significantly lower than that of RANKL combined with miR-143 mimics group and control group (P<0.05).


CONCLUSION
MiR-143 can inhibit the expression of RANK, TRAF6 and downstream NFATc-1 in the RANKL pathway, thereby inhibiting the RANK/RANKL pathway. MiR-143 can inhibit the signal pathway of NF-κB (p65, I-κB-α). MiR-143 inhibits the expression of osteoclast-specific genes TRACP, MMP 9, CtsK and c-Src. That is to say, miR-143 inhibits osteoclast formation by targeting RANK, NF- κB and MAPK signaling pathways.
['He|Xianfeng|X|', 'Zhu|Limei|L|', 'An|Lin|L|', 'Zhang|Jingwei|J|']
D000818:Animals D005786:Gene Expression Regulation D020935:MAP Kinase Signaling System D051379:Mice D035683:MicroRNAs D000072000:NF-KappaB Inhibitor alpha D050778:NFATC Transcription Factors D010010:Osteoclasts D053245:RANK Ligand D000067996:RAW 264.7 Cells D012333:RNA, Messenger D053246:Receptor Activator of Nuclear Factor-kappa B D011994:Recombinant Proteins D015398:Signal Transduction D048029:TNF Receptor-Associated Factor 6 D051996:Transcription Factor RelA
MiR-143 osteoclastogenesis RANK NF-κB MAPK TRAF6 RANK/RANKL