PMID- 36942990 OWN - NLM STAT- MEDLINE DCOM- 20230513 LR - 20230513 IS - 1745-7270 (Electronic) IS - 1672-9145 (Print) IS - 1672-9145 (Linking) VI - 55 IP - 3 DP - 2023 Mar 25 TI - miR-126 mitigates the osteogenic differentiation of human bone marrow-derived mesenchymal stem cells by targeting the ERK1/2 and Bcl-2 pathways. PG - 449-459 LID - 10.3724/abbs.2023016 [doi] AB - Human bone marrow mesenchymal stem cells (hBMMSCs) are a promising cell source for bone engineering owing to their high potential to differentiate into osteoblasts. The objective of the present study is to assess microRNA-126 (miR-126) and examine its effects on the osteogenic differentiation of hBMMSCs. In this study, we investigate the role of miR-126 in the progression of osteogenic differentiation (OD) as well as the apoptosis and inflammation of hBMMSCs during OD induction. OD is induced in hBMMSCs, and matrix mineralization along with other OD-associated markers are evaluated by Alizarin Red S (AR) staining and quantitative PCR (qPCR). Gain- and loss-of-function studies are performed to demonstrate the role of miR-126 in the OD of hBMMSCs. Flow cytometry and qPCR-based cytokine expression studies are performed to investigate the effect of miR-126 on the apoptosis and inflammation of hBMMSCs. The results indicate that miR-126 expression is downregulated during the OD of hBMMSCs. Gain- and loss-of function assays reveal that miR-126 upregulation inhibits the differentiation of hBMMSCs into osteoblasts, whereas the downregulation of miR-126 promotes hBMMSC differentiation, as assessed by the determination of osteogenic genes and alkaline phosphatase activity. Furthermore, the miR-126 level is positively correlated with the production of inflammatory cytokines and apoptotic cell death. Additionally, our results suggest that miR-126 negatively regulates not only B-cell lymphoma 2 (Bcl-2) expression but also the phosphorylation of extracellular signal‑regulated protein kinase (ERK) 1/2. Moreover, restoring ERK1/2 activity and upregulating Bcl-2 expression counteract the miR-126-mediated suppression of OD in hBMMSCs by promoting inflammation and apoptosis, respectively. Overall, our findings suggest a novel molecular mechanism relevant to the differentiation of hBMMSCs into osteoblasts, which can potentially facilitate bone formation by counteracting miR-126-mediated suppression of ERK1/2 activity and Bcl-2 expression. FAU - Zhang, Ying AU - Zhang Y AD - Medical Center of Hip, Luoyang Orthopedic-Traumatological Hospital (Orthopedics Hospital of Henan Province), Luoyang 471002, China. AD - Guangzhou University of Chinese Medicine, Guangzhou 510405, China. FAU - Dong, Yiping AU - Dong Y AD - Henan University of Chinese Medicine, Zhengzhou 450046, China. FAU - Wei, Qiushi AU - Wei Q AD - Institute of Orthopaedics of Guangzhou University of Chinese Medicine, Guangzhou 510240, China. AD - The Third Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510240, China. FAU - Zhuang, Zhikun AU - Zhuang Z AD - Guangzhou University of Chinese Medicine, Guangzhou 510405, China. FAU - Liu, Youwen AU - Liu Y AD - Medical Center of Hip, Luoyang Orthopedic-Traumatological Hospital (Orthopedics Hospital of Henan Province), Luoyang 471002, China. FAU - Yuan, Qiang AU - Yuan Q AD - Henan University of Chinese Medicine, Zhengzhou 450046, China. FAU - He, Wei AU - He W AD - Institute of Orthopaedics of Guangzhou University of Chinese Medicine, Guangzhou 510240, China. AD - The Third Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510240, China. FAU - Jing, Zhenhao AU - Jing Z AD - Henan University of Chinese Medicine, Zhengzhou 450046, China. FAU - Li, Jitian AU - Li J AD - Medical Center of Hip, Luoyang Orthopedic-Traumatological Hospital (Orthopedics Hospital of Henan Province), Luoyang 471002, China. FAU - Li, Peifeng AU - Li P AD - Medical Center of Hip, Luoyang Orthopedic-Traumatological Hospital (Orthopedics Hospital of Henan Province), Luoyang 471002, China. FAU - Zhang, Leilei AU - Zhang L AD - Medical Center of Hip, Luoyang Orthopedic-Traumatological Hospital (Orthopedics Hospital of Henan Province), Luoyang 471002, China. FAU - Hong, Zhinan AU - Hong Z AD - Institute of Orthopaedics of Guangzhou University of Chinese Medicine, Guangzhou 510240, China. AD - The Third Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510240, China. FAU - Zhang, Ning AU - Zhang N AD - Hunan University of Chinese Medicine, Changsha 410208, China. FAU - Wang, Haibin AU - Wang H AD - Guangzhou University of Chinese Medicine, Guangzhou 510405, China. FAU - Li, Wuyin AU - Li W AD - Medical Center of Hip, Luoyang Orthopedic-Traumatological Hospital (Orthopedics Hospital of Henan Province), Luoyang 471002, China. LA - eng PT - Journal Article PL - China TA - Acta Biochim Biophys Sin (Shanghai) JT - Acta biochimica et biophysica Sinica JID - 101206716 RN - 0 (MicroRNAs) RN - 0 (MIRN126 microRNA, human) RN - 0 (BCL2 protein, human) RN - EC 2.7.11.24 (MAPK3 protein, human) RN - EC 2.7.11.24 (MAPK1 protein, human) SB - IM MH - Humans MH - Bone Marrow/metabolism MH - Bone Marrow Cells/metabolism MH - Cell Differentiation/genetics MH - Cells, Cultured MH - Inflammation/metabolism MH - MAP Kinase Signaling System/genetics MH - *Mesenchymal Stem Cells/metabolism MH - *MicroRNAs/metabolism MH - Osteogenesis/genetics PMC - PMC10160225 OTO - NOTNLM OT - Bcl-2 OT - ERK1/2 OT - hBMMSCs OT - miR-126 OT - osteogenic differentiation COIS- The authors declare that they have no conflict of interest. EDAT- 2023/03/22 06:00 MHDA- 2023/05/08 10:17 PMCR- 2023/03/20 CRDT- 2023/03/21 09:13 PHST- 2023/05/08 10:17 [medline] PHST- 2023/03/22 06:00 [pubmed] PHST- 2023/03/21 09:13 [entrez] PHST- 2023/03/20 00:00 [pmc-release] AID - 10.3724/abbs.2023016 [doi] PST - ppublish SO - Acta Biochim Biophys Sin (Shanghai). 2023 Mar 25;55(3):449-459. doi: 10.3724/abbs.2023016.