Department of Stomatology, The First Affiliated Hospital of Jinan University, Guangzhou, Guangdong 510630, P.R. China.
Department of Ophthalmology, First Affiliated Hospital, Guangxi Medical University, Nanning, Guangxi 530021, P.R. China.
Mol Med Rep. 2017 Dec;16(6):9301-9308. doi: 10.3892/mmr.2017.7821. Epub 2017 Oct 19.
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.
经典的 Wnt/β-连环蛋白信号通路在人类间充质干细胞向成骨细胞分化过程中发挥重要作用。越来越多的证据表明，β-连环蛋白的表达部分受到特定 microRNA（miRNA）的调控。本研究旨在探讨 miRNA 在成骨细胞分化中的潜在作用。利用聚合酶链反应（PCR）芯片鉴定分化和未分化牙周膜干细胞（PDLSCs）之间差异表达的 miRNA，并用逆转录-定量 PCR（RT-qPCR）进行验证。由于 miR-214 在 PDLSC 分化过程中明显下调，因此通过沉默和过表达进一步研究其功能。此外，通过茜素红染色和 RT-qPCR 分析碱性磷酸酶（ALP）、骨钙素和骨涎蛋白等成骨分化标志物的 mRNA 表达水平，在诱导后 10 和 21 天评估 PDLSCs 的成骨分化。此外，通过双荧光素酶报告基因检测、RT-qPCR 和 Western blot 分析研究 miR-214 的潜在靶基因，通过 TOPflash/FOPflash 报告质粒系统和 luciferase 检测分析 miR-214 对 Wnt/β-连环蛋白信号通路的影响。本研究结果表明，miR-214 在 PDLSCs 的成骨分化过程中显著下调。值得注意的是，miR-214 的过表达抑制 PDLSC 分化，而其敲低则促进 PDLSC 分化，这可通过成骨细胞特异性基因和 ALP 的 mRNA 表达变化来证实。此外，miR-214 被证实可直接与 CTNNB1 基因的 3'非翻译区相互作用，通过抑制β-连环蛋白抑制 Wnt/β-连环蛋白信号通路。本研究结果表明，miR-214 可能参与调节 Wnt/β-连环蛋白信号通路，并可能作为治疗成骨障碍患者的预防或治疗药物的候选靶点。
Biochem Biophys Res Commun. 2018-6-22
Front Oral Health. 2025-1-31
Stem Cell Res Ther. 2024-11-3
Front Cell Dev Biol. 2024-3-6
Mol Cell Biochem. 2024-9
Front Bioeng Biotechnol. 2022-12-2
Noncoding RNA Res. 2022-11-14
Cell Physiol Biochem. 2016
Cell Physiol Biochem. 2013
Am J Stem Cells. 2013-3-8
Int J Oncol. 2011-12-30
J Tissue Eng Regen Med. 2011-1-12
Biochem Biophys Res Commun. 2010-11-23
Zotero 插件
