Wei Jun-Qiang, Chen Hua, Zheng Xiao-Fei, Zhang Bo-Xun, Wang Yan, Tang Pei-Fu, She Fei, Song Qing, Li Tan-Shi
Department of Orthopedics, General Hospital of PLA, Beijing 100853, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2012 Mar;32(3):291-5.
To study the effect of hsa-miR-654-5p in repressing bone morphogenetic protein 2 (BMP2) mRNA and protein in human bone marrow mesenchymal stem cells (hBMSCs), and explore its regulatory role in osteogenic differentiation of hBMSCs.
hBMSCs in the 4th passage were cultured for 16 h and transfected with hsa-miR-654-5p followed by further culture for 48 h. qRT-PCR and Western blotting were performed to detect the expressions of BMP2 mRNA and protein. Dual-luciferase?reporter gene assay was employed to examine the repression of the BMP2 gene.
BMP2 mRNA and protein expressions were significantly down-regulated in hBMSCs with hsa-miR-654-5p overexpression. Dualluci-ferase reporter gene assay indicated that the predicted target site of BMP2 was repressed directly by hsa-miR-654-5p, but this repression did not occur at the mutant predicted target site of BMP2.
hsa-miR-654-5p can directly repress the mRNA and protein expressions of BMP2 by binding to a specific target site. The changes in hsa-miR-654-5p can play an important role in osteogenic differentiation regulation of hBMSCs.
研究hsa-miR-654-5p对人骨髓间充质干细胞（hBMSCs）中骨形态发生蛋白2（BMP2）mRNA和蛋白的抑制作用，并探讨其在hBMSCs成骨分化中的调控作用。
将第4代hBMSCs培养16 h，转染hsa-miR-654-5p，继续培养48 h。采用qRT-PCR和蛋白质印迹法检测BMP2 mRNA和蛋白的表达。采用双荧光素酶报告基因检测法检测BMP2基因的抑制情况。
hsa-miR-654-5p过表达的hBMSCs中BMP2 mRNA和蛋白表达显著下调。双荧光素酶报告基因检测表明，hsa-miR-654-5p直接抑制BMP2的预测靶位点，但在BMP2的突变预测靶位点未发生这种抑制。
hsa-miR-654-5p可通过与特定靶位点结合直接抑制BMP2的mRNA和蛋白表达。hsa-miR-654-5p的变化在hBMSCs成骨分化调控中起重要作用。
