MicroRNA-214 suppresses osteogenic differentiation of C2C12 myoblast cells by targeting Osterix
MicroRNA-214 suppresses osteogenic differentiation of C2C12 myoblast cells by targeting Osterix
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MicroRNA-214 通过靶向 Osterix 抑制 C2C12 成肌细胞的成骨分化
DOI：
10.1016/j.bone.2013.04.002
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发表时间：
2013-08-01
期刊：
影响因子：
4.1
通讯作者：
Ma, Changyan
中科院分区：
文献类型：
--
作者：
Shi, Kaikai;Lu, Jianlei;Ma, Changyan
Osterix（Osx）是一种成骨细胞特异性转录因子，对成骨细胞分化和骨形成至关重要。Osx基因缺失的小鼠，表现出完全没有骨形成和成骨细胞分化停滞，出生后立即死亡。然而，我们对Osx表达的调控机制的理解仍然很差。微小RNA（miRNA）是一类小型非编码RNA，在细胞和生物体的发育、分化、增殖、生存和肿瘤发生等多种生物过程中发挥关键作用。在这项研究中，我们的目的是研究miRNAs对Osx表达的影响。生物信息学分析预测miR-214可能是Osx的潜在调节因子。使用含有Osx 3' UTR的构建体，通过荧光素酶报告基因测定证明miR-214与Osx 3'非翻译区（3' UTR）的直接结合。缺失突变体的构建显示Osx 3' UTR含有两个miR-214结合位点。在Saos-2和U2 OS细胞中，miR-214表达与Osx表达呈负相关。在Saos-2细胞中强制表达miR-214导致Osx蛋白水平降低。此外，研究了miR-214在C2 C12细胞成骨分化中的作用。我们发现，C2 C12细胞的成骨分化通过下调miR-214表达而增强，如通过增加碱性磷酸酶活性和基质矿化所测量的。综上所述，这些结果表明，miR-214是Osx的一种新的调节剂，并且它作为抑制剂在C2 C12细胞的成骨分化中起重要作用。（C）2013 Elsevier Inc. All rights reserved.
Osterix (Osx) is an osteoblast-specific transcription factor that is essential for osteoblast differentiation and bone formation. Osx-null mice, which exhibit a complete absence of bone formation and arrested osteoblast differentiation, die immediately after birth. However, our understanding of the regulatory mechanism of Osx expression remains poor. MicroRNAs (miRNAs) are a class of small non-coding RNAs that play pivotal Riles in diverse biological processes, including the development, differentiation, proliferation, survival, and oncogenesis of cells and organisms. In this study, we aimed to investigate the impact of miRNAs on Osx expression. Bioinformatic analyses predicted that miR-214 would be a potential regulator of Osx. The direct binding of miR-214 to the Osx 3' untranslated region (3' UTR) was demonstrated by a luciferase reporter assay using a construct containing the Osx 3' UTR. Deletion mutant construction revealed that the Osx 3' UTR contained two miR-214 binding sites. MiR-214 expression was inversely correlated with Osx expression in Saos-2 and U2OS cells. The forced expression of miR-214 in Saos-2 cells led to a reduction in the level of Osx protein. Moreover, the role of miR-214 in the osteogenic differentiation of C2C12 cells was investigated. We found that the osteogenic differentiation of C2C12 cells was enhanced by the downregulation of miR-214 expression, as measured by increased alkaline phosphatase activity and matrix mineralization. Taken together, these results indicate that miR-214 is a novel regulator of Osx, and that it plays an important role in the osteogenic differentiation of C2C12 cells as a suppressor. (C)2013 Elsevier Inc. All rights reserved.