miR-214 promotes periodontal ligament stem cell osteoblastic differentiation by modulating Wnt/β‑catenin signaling.
miR-214 promotes periodontal ligament stem cell osteoblastic differentiation by modulating Wnt/β‑catenin signaling.
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miR-214通过调节Wnt/β-蛋白信号传导来促进牙周韧带干细胞成骨细胞分化。
DOI：
10.3892/mmr.2017.7821
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发表时间：
2017-12
影响因子：
3.4
通讯作者：
Feng Z
中科院分区：
文献类型：
--
作者：
Cao F;Zhan J;Chen X;Zhang K;Lai R;Feng Z
典型的Wnt/β-catenin信号在人间充质干细胞向成骨细胞的分化过程中起重要作用。越来越多的证据表明，β-catenin的表达在一定程度上是由特定的microrna （mirna）调节的。本研究的目的是探讨mirna在成骨细胞分化中的可能作用。采用聚合酶链反应（PCR）阵列鉴定分化和未分化牙周韧带干细胞（PDLSCs）之间差异表达的mirna，并采用逆转录定量PCR （RT-qPCR）进行验证。由于miR-214在PDLSC分化过程中被发现显著下调，因此通过沉默和过表达进一步研究其功能。此外，在诱导后10和21天，采用茜素红染色和RT-qPCR分析成骨分化标志物碱性磷酸酶（ALP）、骨钙素和骨涎蛋白的mRNA表达水平，评估PDLSCs的成骨分化情况。此外，使用双荧光素酶报告基因检测、RT-qPCR和western blot分析研究了miR-214的潜在靶基因，而使用TOPflash/FOPflash报告基因质粒系统和荧光素酶检测来检测miR-214对Wnt/β-catenin信号传导的影响。目前的结果表明，miR-214在PDLSCs的成骨细胞分化过程中显著下调。值得注意的是，它的过表达抑制了PDLSC的分化，而它的低表达促进了PDLSC的分化，这是由成骨特异性基因和ALP mRNA表达的改变所揭示的。此外，miR-214被证明直接与β-catenin基因CTNNB1的3 ' -非翻译区相互作用，并通过抑制β-catenin抑制Wnt/β-catenin信号传导。本研究结果提示，miR-214可能参与Wnt/β-catenin信号通路的调控，可能有潜力作为候选靶点开发用于治疗成骨疾病患者的预防或治疗药物。
The canonical Wnt/β-catenin signaling is important in the differentiation of human mesenchymal stem cells into osteoblasts. Accumulating evidence suggests that the expression of β-catenin is, in part, regulated by specific microRNAs (miRNAs). The aim of the present study was to investigate the putative roles of miRNAs in osteoblast differentiation. Polymerase chain reaction (PCR) arrays were used to identify miRNAs that were differentially expressed between differentiated and non-differentiated periodontal ligament stem cells (PDLSCs), and reverse transcription-quantitative PCR (RT-qPCR) was used for validation. Since miR-214 was revealed to be significantly downregulated during PDLSC differentiation, its function was further investigated via silencing and overexpression. In addition, osteogenic differentiation of PDLSCs was evaluated at 10 and 21 days following induction, using Alizarin red staining and RT-qPCR analysis for mRNA expression levels of the osteogenic differentiation markers alkaline phosphatase (ALP), osteocalcin and bone sialoprotein. Furthermore, the potential target genes of miR-214 were investigated using a dual-luciferase reporter assay, RT-qPCR and western blot analysis, whereas a TOPflash/FOPflash reporter plasmid system followed by a luciferase assay was used to examine the effects of miR-214 on Wnt/β-catenin signaling. The present results demonstrated that miR-214 was significantly downregulated during the osteoblastic differentiation of PDLSCs. Notably, its overexpression inhibited PDLSC differentiation, whereas its knockdown promoted PDLSC differentiation, as revealed by alterations in mRNA expression of osteoblast-specific genes and ALP. In addition, miR-214 was demonstrated to directly interact with the 3′-untranslated region of the β-catenin gene CTNNB1, and suppressed Wnt/β-catenin signaling through the inhibition of β-catenin. The results of the present study suggested that miR-214 may participate in the regulation of the Wnt/β-catenin signaling pathway, and may have potential as a candidate target for the development of preventive or therapeutic agents for the treatment of patients with osteogenic disorders.
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影响因子：
4.5
作者：
Shang, Y. C.;Zhang, C.;Li, Y.
通讯作者：
Li, Y.
DOI：
10.1016/j.bbrc.2010.08.031
发表时间：
2010-11-12
影响因子：
3.1
作者：
Wang, Tao;Xu, Zhengyu
通讯作者：
Xu, Zhengyu
影响因子：
7.6
作者：
Huang GT;Gronthos S;Shi S
通讯作者：
Shi S
影响因子：
16
作者：
Jones-Rhoades, MW;Bartel, DP
通讯作者：
Bartel, DP
影响因子：
6.5
作者：
Feng, Jue;Iwama, Atsushi;Kohu, Kazuyoshi
通讯作者：
Kohu, Kazuyoshi