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miR-181a Reguates the Levels of OPG Protein and Its Effect on the Survival of Osteoclast
Gao Jing1,2,3, Shao Bingyi1,2,3*
1Department of Endodontics, Affiliated Stomatological Hospital, Chongqing Medical University, Chongqing 401147, China;
2Chongqing Key Laboratory of Oral Diseases and Biomedical Sciences, Chongqing 401147, China;
3Chongqing Municipal Key Laboratory of Oral Biomedical Engineering of Higher Education, Chongqing 401147, China
2Chongqing Key Laboratory of Oral Diseases and Biomedical Sciences, Chongqing 401147, China;
3Chongqing Municipal Key Laboratory of Oral Biomedical Engineering of Higher Education, Chongqing 401147, China
Abstract: The research group predict and verify the target molecule osteoprotegerin (OPG) mRNA of miR-181a in BMMSCs and investigate the effect of OPG on osteoclast activity in the pathogenesis of osteoporosis. The putative target mRNA of miR-181a were predicted by bioinformatics approach. The dual luciferase vector containing 3′ untranslated regions (3′UTR) of OPG mRNA was constructed， and the 3′UTR was regarded as the binding site of miR-181a. The interaction between miR-181a and target mRNA was verified by dual luciferase activity analysis. miR-181a was transfected into BMMSCs and co-cultured with osteoclast， and TRAP staining was used to detect the number of osteoclast in co-culture system. The results of dual luciferase reporter gene assay indicated that miR-181a regulated OPG by binding to the 3′UTR of OPG mRNA. In vitro co-culture assays confirmed that BMMSCs transfected with an miR-181a/mimics increased osteoclast number， whereas BMMSCs/inhibitor reduced osteoclast number. We concluded that miR-181a down-regulates the level of OPG protein which might be an important factor to affect the activity of osteoclast.
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