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MiR‐17‐3p inhibits osteoblast differentiation by downregulating Sox6 expression

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Osteoporosis and osteoarthritis are orthopedic disorders that affect millions of elderly people worldwide; stimulation of bone formation is a potential therapeutic strategy for the treatment of these conditions. As the… Click to show full abstract

Osteoporosis and osteoarthritis are orthopedic disorders that affect millions of elderly people worldwide; stimulation of bone formation is a potential therapeutic strategy for the treatment of these conditions. As the only bone‐forming cells, osteoblasts play a key role in bone reconstruction. The microRNA miR‐17‐3p is downregulated during osteogenic differentiation of human bone marrow mesenchymal stem cells, but its precise role in this process is unknown. Here, we investigated the role of miR‐17‐3p in osteoblast differentiation. An in vitro model of osteogenesis was established by treating MC3T3‐E1 murine preosteoblast cells with bone morphogenetic protein 2 (BMP2). The expression of miR‐17‐3p in BMP2‐induced MC3T3‐E1 cells was detected by reverse transcription‐quantitative PCR, and its effects on cells transfected with miR‐17‐3p mimic or inhibitor were evaluated by Alizarin Red staining, alkaline phosphatase (ALP) activity assay, and by detection of osteoblast markers including the ALP, collagen type I α1 chain, and osteopontin genes. Bioinformatics analysis was carried out to identify putative target genes of miR‐17‐3p, and the luciferase reporter assay was used for functional validation. Rescue experiments were performed to determine whether SRY‐box transcription factor 6 (Sox6) plays a role in the regulation of osteoblast differentiation by miR‐17‐3p. We report that miR‐17‐3p was downregulated upon BMP2‐induced osteoblast differentiation in MC3T3‐E1 cells, and this was accompanied by decreased differentiation and mineralization, ALP activity, and expression of osteogenesis‐related genes. Sox6 was confirmed to be a target gene of miR‐17‐3p in osteoblasts, and the inhibitory effect of miR‐17‐3p on osteoblast differentiation was observed to occur via Sox6. These results suggest the existence of a novel mechanism underlying miRNA‐mediated regulation of osteogenesis, which has potential implications for the treatment of orthopedic disorders.

Keywords: mir; differentiation; role; osteoblast differentiation; expression mir

Journal Title: FEBS Open Bio
Year Published: 2020

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