长链非编码RNA MEG3的下调通过miR-27a-3p/IGF1轴抑制牙周炎中牙周膜干细胞（PDLSCs）的成骨分化。
Down-regulation of long non-coding RNA MEG3 suppresses osteogenic differentiation of periodontal ligament stem cells (PDLSCs) through miR-27a-3p/IGF1 axis in periodontitis.
作者信息
Liu Yi, Liu Chunpeng, Zhang Ankui, Yin Shichang, Wang Ting, Wang Yan, Wang Meiming, Liu Yixin, Ying Qiaohui, Sun Jinrui, Wei Fulan, Liu Dongxu, Wang Chunling, Ge Shaohua
机构信息
Shandong Provincial Key Laboratory of Oral Tissue Regeneration, School of Stomatology, Shandong University, Jinan, Shandong 250012, China.
Department of Orthodontics, School of Stomatology, Shandong University, Jinan, Shandong 250012, China.
出版信息
Aging (Albany NY). 2019 Aug 9;11(15):5334-5350. doi: 10.18632/aging.102105.
OBJECTIVE
This study aimed to investigate the roles of long noncoding RNA (lncRNA) maternally expressed gene 3 (MEG3) in osteogenic differentiation of periodontal ligament stem cells (PDLSCs) in periodontitis.
METHODS
Differentially expressed lncRNAs and mRNAs between periodontitis periodontal ligament tissues and healthy periodontal ligament tissues were selected out using R project. PDLSCs were identified using flow cytometry. Western blot was employed to detect pathway relative proteins. Besides, targeted relationships between lncRNA and miRNA, as well as miRNA and mRNA were verified by dual luciferase reporter gene assay. Osteogenic differentiation of PDLSCs was assessed by alkaline phosphatase (ALP) staining and Alizarin Red Staining (ARS). Markers for osteoblast (Runx2, Osterix, Osteocalcin, Colla1) were detected using western blot.
RESULTS
LncRNA MEG3 and were both down-regulated, while miR-27a-3p was up-regulated in periodontitis samples compared with healthy samples. Overexpression of MEG3 promoted osteogenic differentiation by enhancing expression of yet suppressing expression of miRNA-27a-3p. Meanwhile, the results of ALP and ARS staining indicated that up-regulation of lncRNA MEG3 or promoted osteogenic differentiation in PDLSCs, which could be reversed with up-regulation of miRNA-27a-3p.
CONCLUSION
Down-regulation of MEG3 suppressed osteogenic differentiation of PDLSCs through miR-27a-3p/ axis in periodontitis.
目的
本研究旨在探讨长链非编码RNA（lncRNA）母源表达基因3（MEG3）在牙周炎中牙周膜干细胞（PDLSCs）成骨分化中的作用。
方法
使用R软件筛选出牙周炎牙周膜组织与健康牙周膜组织之间差异表达的lncRNAs和mRNAs。采用流式细胞术鉴定PDLSCs。运用蛋白质印迹法检测通路相关蛋白。此外，通过双荧光素酶报告基因检测验证lncRNA与miRNA以及miRNA与mRNA之间的靶向关系。通过碱性磷酸酶（ALP）染色和茜素红染色（ARS）评估PDLSCs的成骨分化。使用蛋白质印迹法检测成骨细胞标志物（Runx2、Osterix、骨钙素、Colla1）。
结果
与健康样本相比，lncRNA MEG3和 在牙周炎样本中均下调，而miR-27a-3p上调。MEG3的过表达通过增强 的表达同时抑制miRNA-27a-3p的表达来促进成骨分化。同时，ALP和ARS染色结果表明，lncRNA MEG3或 的上调促进了PDLSCs的成骨分化，而miRNA-27a-3p的上调可逆转这一过程。
结论
在牙周炎中，MEG3的下调通过miR-27a-3p/ 轴抑制了PDLSCs的成骨分化。
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