Satb2 expression in Foxc1-promoted osteogenic differentiation of MC3T3-E1 cells is negatively regulated by microRNA-103-3p
Satb2 expression in Foxc1-promoted osteogenic differentiation of MC3T3-E1 cells is negatively regulated by microRNA-103-3p
复制标题
Foxc1促进MC3T3-E1细胞成骨分化中的Satb2表达受到microRNA-103-3p的负调节
DOI：
10.1093/abbs/gmz037
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发表时间：
2019-06-01
影响因子：
3.7
通讯作者：
Shen, Guofang
中科院分区：
文献类型：
--
作者：
Shen, Hongzhou;Lu, Chenpei;Shen, Guofang
叉头转录因子C1 （Foxc1）是控制颅骨发育和成骨分化的细胞命运决定因子。此前研究表明，多种microRNAs （miRNAs）在成骨过程中发挥重要作用，调控成骨分化的复杂过程。然而，在foxc1促进成骨分化过程中，miRNA表达的变化机制尚不清楚。在本研究中，我们成功地在MC3T3-E1细胞中过表达Foxc1基因，并利用miRNA芯片研究了成骨诱导后第3天miRNA表达谱的变化。发现9个下调的mirna和8个上调的mirna存在差异表达。在这些mirna中，miR-103-3p在foxc1过表达的MC3T3-E1细胞中持续下调，并通过功能获得和功能丧失试验被鉴定为成骨分化的负调节因子。特异的富含at的序列结合蛋白2 （Satb2）是一种关键的成骨转录因子，被鉴定为miR-103-3p的靶向基因，并通过实时聚合酶链反应、western blot分析和荧光素酶测定进行验证。过表达miR-103-3p可显著抑制Satb2的表达，减弱foxc1促进的成骨分化。综上所述，我们的研究结果阐明了foxc1促进成骨分化早期MC3T3-E1细胞的miRNA表达谱，并提示miR-103-3p通过直接靶向Satb2对MC3T3-E1细胞的成骨分化起到负调控作用。
The forkhead transcription factor C1 (Foxc1) is a cell-fate-determining factor that controls cranial bone development and osteogenic differentiation. Previously, it was demonstrated that various microRNAs (miRNAs) play important roles in osteogenesis and regulate the complex process of osteogenic differentiation. However, it remains unclear how miRNA expression changes during Foxc1-promoted osteogenic differentiation. In this study, we successfully overexpressed the Foxc1 gene in MC3T3-E1 cells and investigated the alterations in the miRNA expression profile on day 3 after osteogenic induction by using a miRNA microarray. Nine downregulated miRNAs and eight upregulated miRNAs were found to be differentially expressed. Among these miRNAs, miR-103-3p was consistently downregulated in the Foxc1-overexpressing MC3T3-E1 cells and was identified as a negative regulator of osteogenic differentiation by using a gain-and lose-of-function assay. The special AT-rich sequence-binding protein 2 (Satb2), a pivotal osteogenic transcription factor, was identified as the miR-103-3p targeting gene and was verified by real-time polymerase chain reaction, western blot analysis, and luciferase assay. Overexpression of miR-103-3p markedly inhibited the expression of Satb2 and attenuated Foxc1-promoted osteogenic differentiation. Taken together, our results elucidated the miRNA expression profiles of MC3T3-E1 cells in the early stage of Foxc1-promoted osteogenic differentiation and suggested that miR-103-3p acts as a negative regulator of the osteogenic differentiation of MC3T3-E1 cells by directly targeting Satb2.