RESUMO
AIMS: The study aimed to investigate the roles of miR-483-5p and IGF2 in osteoclast formation. METHODS: Blood and bone tissues were collected from osteoporosis and non-osteoporosis patients with hip fractures for gene expression analysis. CD14 + peripheral blood mononuclear cells (PBMCs) were isolated for differentiating osteoclasts. MiR-483-5p mimic and inhibitor was transfected into CD14 + PBMCs, respectively. Predicted by TargetScan and verified by Dual-luciferase reporter assay system, insulin-like growth factor-2 (IGF2) could be targeted by miR-483-5p. IGF2 expression vector was co-transfected with miR-483-5p mimic to study the role of IGF2 in miR-483-5p affecting osteoclast differentiation. Flow cytometry was performed for cell apoptosis analysis. RESULTS: High-expressed miR-483-5p and low-expressed IGF2 were frequently found in the serums and bone tissues derived from osteoporotic patients. We found that up-regulation of miR-483-5p in CD14 + PBMCs notably increased the number of TRAP-positive cells, at the same time, the expression levels of TRAP, nuclear factor of activated T-cells (NFATc1), cytoplasmic 1 (NFAT2) and Cathepsin K (CTSK) were also up-regulated. However, overexpressed IGF2 effectively reversed such effects produced by up-regulation of miR-483-5p on osteoclastogenesis-related factors in CD14 + PBMCs. Moreover, forced expression of IGF2 could also enhance apoptosis of osteoclasts reduced by miR-483-5p. CONCLUSIONS: Our study suggests that miRNA-483-5p is involved in the pathogenesis of osteoporosis by promoting osteoclast differentiation.
