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  • Title: [Changes of microRNA and target gene expression levels in osteogenic differentiation of human bone marrow mesenchymal stem cells].
    Author: Wei J, Zhang B, Zheng X, Chen H, Tian X, Tang P, Song Q, Li T.
    Journal: Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi; 2012 Apr; 26(4):483-8. PubMed ID: 22568334.
    Abstract:
    OBJECTIVE: To clarify the trends of expression levels of several up-regulated micro RNA (miRNA) in tissues of atrophic bone nonunion and mRNAs and proteins of their related target genes in osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs), and to explore their biological functions. METHODS: The hBMSCs were isolated from bone marrow of iliac bone by gradient centrifugation, and cultured. Osteogenic culture medium was used for osteogenic differentiation of the 4th generation of hBMSCs. The changes of corresponding miRNAs, mRNA and protein expression levels of related target genes were observed at 0 hour, 12 hours, 1 day, 2 days, 4 days, 7 days, and 14 days, by quantitative real-time PCR and Western blot. RESULTS: In the process of hBMSCs osteogenic differentiation, the mRNA and protein expression levels of osteoblastic target genes [alkaline phosphatase liver/bone/kidney (ALPL), bone morphogenetic protein 2 (BMP-2), and platelet-derived factor alpha polypeptide (PDGF-A)] at most time points increased significantly when compared with the values at 0 hour except that of BMP-2 decreased at 12 hours and 1 day, with maximum changes at 1 to 7 days. The miRNA expression levels, mRNA and protein expression levels changed significantly at different time points, while the trends of hsa-miRNA-149 and hsa-miRNA-654-5p changes were negatively correlated with the trends of ALPL and BMP-2 mRNA and protein expression changes respectively (P < 0.05). There was no obviously negative correlation between the trends of hsa-miRNA-221 change and PDGF-A change (P > 0.05). CONCLUSION: In the osteogenic differentiation process of hBMSCs, hsa-miRNA-149* and hsa-miRNA-654-5p are closely related with the mRNA and protein regulation of ALPL and BMP-2, respectively.
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