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Abstract  Sustained hypoxia inhibits osteogenesis and osteoblast differentiation by downregulating the expression of runt‐related transcription factor 2 (Runx2). MicroRNAs (miRNAs) have been shown to regulate osteogenesis and osteoblast differentiation. In the present study, we profiled miRNAs, with microRNA array and quantitative real‐time polymerase chain reaction (RT‐PCR) methods, in mouse osteoblast (MC3T3‐E1) cells under hypoxia. Then, we investigated regulation by miRNA‐21‐5p on the expression of Runx2 and other osteoblast differentiation‐associated markers via gain‐of‐function and loss‐of‐function strategies. We found that expression of miRNA‐21‐5p, miRNA‐210‐5p, and other eight miRNAs was upregulated significantly in hypoxia‐treated MC3T3‐E1 cells. miRNA‐21‐5p overexpression downregulated the expression of the mRNA and protein of suppressor of mothers against decapentaplegic ( SMAD7 ) markedly, the 3′‐untranslated region (3′‐UTR) of which was highly homologous with the miRNA‐21‐5p sequence. miRNA‐21‐5p overexpression upregulated the protein expression of Runx2 in hypoxia‐treated MC3T3‐E1 cells, although mRNA expression of Runx2 and other osteoblast differentiation‐associated molecules (eg, osteocalcin, procollagen type 1 amino‐terminal propeptide, P1NP) were not regulated by it; such upregulation was SMAD7‐dependent. In conclusion, hypoxia‐responsive miRNA‐21‐5p promoted Runx2 expression (at least in part) by targeting the 3′‐UTR and downregulating  SMAD7  expression. Our study suggests a protective role of miRNA‐21‐5p in promoting osteoblast differentiation under hypoxia.

Hypoxia‐responsive miRNA‐21‐5p inhibits Runx2 suppression by targeting SMAD7 in MC3T3‐E1 cells