THE EXPRESSION OF MIR-153-3P REGULATES THE OSTEOGENIC DIFFERENTIATION OF BONE MARROW DERIVED MESENCHYMAL STEM CELLS BY TARGETING RUNX2

Rong Ma^{1, 2}, Junyi Miao³, Shuwen Fang³, Mufan Ji³, Xin yi^{1, *}

¹Department of Human Anatomy, Medical School, Nantong University, Nantong 226001, Jiangsu, China - ²Department of Orthopedics, Nantong University Affiliated Jianhu People's Hospital, Jianhu 224799, Jiangsu, China - ³Department of Clinical Medicine, Medical School, Nantong University, Nantong 226001, Jiangsu, China

Objective: The study aimed to explore the effect of miR-153-3p on the osteogenic differentiation of human bone marrow derived mesenchymal stem cells (hBMSCs) and confirm its targeting effect on RUNX2. 

Methods: The expression of miR-153-3p in the osteogenic differentiation of hBMSCs was detected by qRT-PCR. The hBMSCs were transfected with miR-153-3p mimic or inhibitor, and then its viability was analyzed with cell counting kit-8, proliferation ability was analyzed with Ki67 immunofluorescence and osteogenic differentiation was observed by Alizarin red staining, alkaline phosphatase (ALP) activity assay and Western blot for RUNX2 and Collagen I. The miR-153-3p targeting effect on RUNX2 was verified by luciferase reporter gene experiment. 

Results: During the osteogenic differentiation of hBMSCs, the expression of miR-153-3p decreased at 1 d and reached a low peak at 3 d. After miR-153-3p mimic or inhibitor transfection, miRNA-153-3p expression level of hBMSCs dramatically increased or decreased, but the cell viability and proliferation ability were not notably changed. After miR-153-3p inhibitor transfection, the osteogenic differentiation was notably promoted which was confirmed with Alizarin red staining, ALP activity and Western blot for RUNX2 and Collagen I, while miR-153-3p mimic transfection, the osteogenic differentiation was notably suppressed. The result of luciferase reporter gene experiment verified that RUNX2 was the target gene of miR-153-3p. 

Conclusion: The expression of miR-153-3p regulates the osteogenic differentiation of hBMSCs by targeting RUNX2.