miR-3960对成骨细胞分化的影响

Effect of miR-3960 on osteogenic differentiation

背景:成骨细胞的分化成熟过程涉及多种激素和细胞因子对成骨细胞分化相关基因表达的调控,近来发现多个微小RNAs也参与了这一调控过程。目的:观察miR-3960在小鼠骨髓基质细胞向成骨细胞分化过程中的作用。方法:将miR-3960表达载体pSilencer4.1-miR-3960转染骨髓基质细胞,构建miR-3960过表达细胞模型,随后予300μg/L骨形态发生蛋白2诱导分化,观察成骨细胞分化指标变化。结果与结论:转染pSilencer4.1-miR-3960能够在细胞中稳定地高表达miR-3960。miR-3960过表达促进骨髓基质细胞向成骨细胞分化过程中的碱性磷酸酶活性增高和骨钙素分泌,增加细胞中的钙沉积量。抑制miR-3960降低骨髓基质细胞向成骨细胞分化过程中的碱性磷酸酶活性,减少骨钙素分泌,降低钙沉积量。表明miR-3960可以促进成骨细胞分化。

BACKGROUND：Osteoblast differentiation involves highly regulated processes mediated by a large number of hormones and transcription factors.Emerging evidence reveals an additional level of regulation that is mediated by microRNAs.miR-3960 is preferentially expressed in osteoblasts,which suggests that miR-3960 may take part in the regulation process of osteogenic differentiation.OBJECTIVE：To investigate the role of miR-3960 in osteogenic differentiation of primary mouse bone marrow stromal cells.METHODS：The parameters of osteoblast differentiation were measured after transfection of bone marrow stromal cells with pSilencer4.1-miR-3960.Osteoblastic differentiation was induced by the addition of 300 μg/L bone morphogenetic protein 2.RESULTS AND CONCLUSION：Stable and high expression of miR-3960 was obtained in the cells after transfection with pSilencer4.1-miR-3960.Compared with control cells,the levels of alkaline phosphatase activity,osteocalcin secretion and calcium deposition were increased by transfection of pSilencer4.1-miR-3960.Treatment with anti-miR-3960 attenuated the levels of alkaline phosphatase activity and osteocalcin secretion as well as reduced calcium deposition.All these results suggest that miR-3960 promotes osteoblast differentiation of bone marrow stromal cells.