Shandong Provincial Key Laboratory of Oral Tissue Regeneration, School of Stomatology, Shandong University, Jinan, China.
Department of Orthodontics, School of Stomatology, Shandong University, Jinan, China.
J Cell Physiol. 2019 Apr;234(4):4617-4626. doi: 10.1002/jcp.27248. Epub 2018 Sep 7.
This study aims to discuss long noncoding RNA (lncRNA) maternally expressed gene 3 (MEG3) function of regulating osteogenesis in human periodontal ligament cells (hPDLCs).
First, use of a mineralizing solution induced osteogenic differentiation of hPDLCs to establish a differentiated cell model. Through microarray analysis, we selected a lncRNA MEG3 with marked changes between differentiated and undifferentiated cells. The quantitative polymerase chain reaction was used to detect the MEG3 content and an enzyme-linked immunosorbent assay was used to detect changes in related proteins. Cell viability was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and apoptosis was measured by flow cytometry. Alizarin red staining was also used to evaluate cells' osteogenic level. Finally, RNA-binding protein immunoprecipitation assays were conducted to further clarify the endogenous relationship between MEG3 and bone morphogenetic protein 2 ( BMP2) in hPDLCs.
MEG3 was downregulated in osteogenic differentiation hPDLCs induced by mineralizing solution. Overexpression of MEG3 inhibited cell viability and increased cell apoptosis. MEG3 overexpression can reverse osteogenic differentiation induced by mineralizing solution. MEG3 can suppress BMP2 through interaction with heterogeneous nuclear ribonucleoprotein I.
Upregulation of MEG3 inhibits the osteogenic differentiation of periodontal ligament cells by downregulating BMP2 expression.
本研究旨在探讨长非编码 RNA（lncRNA）母系表达基因 3（MEG3）在人牙周膜细胞（hPDLCs）成骨中的调控作用。
首先，使用矿化液诱导 hPDLCs 成骨分化，建立分化细胞模型。通过微阵列分析，我们选择了一个在分化和未分化细胞之间变化明显的 lncRNA MEG3。采用定量聚合酶链反应检测 MEG3 含量，采用酶联免疫吸附试验检测相关蛋白变化。采用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐法检测细胞活力，采用流式细胞术检测细胞凋亡。茜素红染色法也用于评估细胞的成骨水平。最后，通过 RNA 结合蛋白免疫沉淀试验进一步阐明 hPDLCs 中 MEG3 与骨形态发生蛋白 2（BMP2）之间的内源性关系。
矿化液诱导的成骨分化 hPDLCs 中 MEG3 表达下调。MEG3 过表达抑制细胞活力并增加细胞凋亡。MEG3 过表达可逆转矿化液诱导的成骨分化。MEG3 通过与异质核核糖核蛋白 I 相互作用抑制 BMP2。
上调 MEG3 通过下调 BMP2 表达抑制牙周膜细胞的成骨分化。
