The function of miR-4666a-3p in the diagnosis of osteoporotic fractures and the promotion of fracture healing.

INTRODUCTION AND

OBJECTIVE

MicroRNAs (miRNAs) regulate osteogenic differentiation in osteoporotic fractures (OF). The aim of the study is to investigate the diagnostic value and regulatory role of miR-4666a-3p in OF.

MATERIAL AND

METHODS

The study is based on clinical samples (89 healthy controls, 79 OP patients, 83 OF patients) and hFOB1.19 cell experiments. RT-qPCR analyzed miR-4666a-3p expression, and logistic regression identified OF risk factors. ELISA measured RANKL and OPG concentrations. In vitro, osteoblast differentiation was induced to assess ALP activity, Runx2, Osteocalcin, Col1a1, cell viability (CCK-8), and apoptosis (flow cytometry). Target genes were bioinformatically predicted and confirmed via dual-luciferase assay. Rescue experiments explored the role of GSK3B in miR-4666a-3p-mediated osteogenic regulation. All experiments were repeated at least three times.

RESULTS

Downregulation of miR-4666a-3p could distinguish healthy controls and OP patients, further differentiate between OP and OF patients, and may predict the risk of OF occurrence. During osteoblast differentiation, miR-4666a-3p expression increased, ALP activity elevated, and key marker levels upregulated. Overexpression of miR-4666a-3p promoted osteoblast differentiation and activity, and reduced apoptosis rate, whereas inhibiting miR-4666a-3p had the opposite effect. MiR-4666a-3p negatively regulated the downstream target GSK3B. Overexpression of GSK3B reversed the osteogenic effects of miR-4666a-3p.

CONCLUSIONS

Downregulation of miR-4666a-3p may predict OF risk. Its upregulation promoted osteoblast differentiation by targeting GSK3B, potentially facilitating fracture healing.