MicroRNA-145 suppresses osteogenic differentiation of human jaw bone marrow mesenchymal stem cells partially via targeting semaphorin 3A
MicroRNA-145 suppresses osteogenic differentiation of human jaw bone marrow mesenchymal stem cells partially via targeting semaphorin 3A
复制标题
MicroRNA-145部分通过靶向semaphorin 3A抑制人颌骨骨髓间充质干细胞的成骨分化
DOI：
10.1080/03008207.2019.1643334
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发表时间：
2019-07-28
影响因子：
2.9
通讯作者：
Ma, Changyan
中科院分区：
文献类型：
--
作者：
Jin, Yucui;Hong, Fangling;Ma, Changyan
目的：人颌骨骨髓间充质干细胞(h-JBMMSCs)是一种具有成骨分化潜能的多潜能前体细胞。MicroRNAs(MiRNAs)是成骨细胞分化的重要调节因子。本研究旨在探讨miR-145及其靶蛋白在h-JBMMSCs成骨分化中的作用。材料和方法：分离h-JBMMSCs，在成骨介质中培养，分别用miR-145模拟物和抑制剂上调和抑制miR-145的表达。碱性磷酸酶和茜素红S染色检测成骨分化，实时定量逆转录聚合酶链式反应检测成骨标志物。结果：miR-145基因在h-JBMMSCs成骨过程中表达下调。抑制miR-145可促进h-JBMMSCs的成骨分化，表现为碱性磷酸酶(ALP)活性增强，矿化增强，成骨标志物如Run-Related转录因子2(RUNX2)、Osterix(OSX)、ALP和COL1A1的表达水平增加。miR-145可以负向调节信号素3A(SEMA3A)，而信号素3A(SEMA3A)是成骨的正向调节因子。结论：miR-145直接靶向SEMA3A，并提示miR-145作为抑制因子在h-JBMMSCs的成骨分化中起重要作用。
Purpose: Human jaw bone marrow mesenchymal stem cells (h-JBMMSCs) are multipotent progenitor cells with osteogenic differentiation potential. MicroRNAs (miRNAs) have emerged as crucial modulators of osteoblast differentiation. In this study, we focus on the role ofmiR-145and its target protein in osteoblast differentiation of h-JBMMSCs.Materials and Methods: h-JBMMSCs were isolated and cultured in osteogenic medium.miR-145mimics and inhibitors were used to elevate and inhibitmiR-145expression, respectively. Osteogenic differentiation was determined by Alkaline phosphatase (ALP) and Alizarin red S (ARS) staining, and osteogenic marker detection using quantitative real-time reverse transcription PCR (qRT-PCR) assay. Bioinformatic analysis and luciferase reporter assay were used to identify the target gene ofmiR-145.Results:MiR-145was down-regulated during osteogenesis of h-JBMMSCs. Inhibition ofmiR-145promoted osteogenic differentiation of h-JBMMSCs, revealed by enhanced activity of alkaline phosphatase (ALP), greater mineralisation, and increased expression levels of the osteogenic markers, such as Runt-related transcription factor 2 (RUNX2), Osterix (OSX), ALP and COL1A1.MiR-145could negatively regulate semaphorin3A (SEMA3A), which acts as a positive regulator of osteogenesis.MiR-145inhibitor induced osteogenesis could be partially attenuated by SEMA3A siRNA treatment in h-JBMMSCs.Conclusions: Our data show thatmiR-145directly targets SEMA3A, and also suggestmiR-145as a suppressor, plays an important role in the osteogenic differentiation of h-JBMMSCs.