State Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shaanxi Clinical Research Center for Oral Diseases, Department of Orthodontics, School of Stomatology, Fourth Military Medical University, Xi'an, China.
Oral Dis. 2024 Apr;30(3):1322-1336. doi: 10.1111/odi.14467. Epub 2023 Jan 13.
This study aimed to investigate the mechanism of N6-methyladenosine (m6A) in the osteogenic differentiation of periodontal ligament stem cells (PDLSCs) from periodontitis patients.
Differentially m6A-methylated lncRNA/mRNA profiles were detected by a m6A epitranscriptomic microarray. Bioinformatics analysis was performed by Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analysis. The transfection efficiency of the lentivirus was detected. The osteogenic activity of PDLSCs from periodontitis patients (PPDLSCs) was assessed.
The microarray results showed that 275 lncRNAs and 1292 mRNAs were significantly differentially methylated between PPDLSCs and PDLSCs from healthy people. Among those lncRNAs, lncRNA4114 (transcript_ID: ENST00000444114) showed both reduced m6A methylation levels and expression levels in PPDLSCs. Further bioinformatics analysis predicted that the differentially methylated mRNAs were mainly involved in cell interaction, stem cell pluripotency, and osteogenic differentiation signals. Then, overexpression of methyltransferase like 3 (METTL3) promoted the osteogenic differentiation of PPDLSCs, while knocking down METTL3 showed an inhibitory effect. Furthermore, METTL3 overexpression promotes the stability of lncRNA4114 to upregulate the expression level. Moreover, lncRNA4114 overexpression promoted the osteogenic differentiation of PPDLSCs.
METTL3 promotes the osteogenic differentiation of PPDLSCs by regulating the stability of lncRNA4114.
本研究旨在探讨 N6-甲基腺苷（m6A）在牙周炎患者牙周韧带干细胞（PDLSCs）成骨分化中的作用机制。
采用 m6A 转录组芯片检测差异 m6A 甲基化长链非编码 RNA/mRNA 谱。通过基因本体论和京都基因与基因组百科全书通路分析进行生物信息学分析。检测慢病毒的转染效率。评估牙周炎患者来源的 PDLSCs（PPDLSCs）的成骨活性。
芯片结果显示，PPDLSCs 和健康人来源的 PDLSCs 之间有 275 个 lncRNA 和 1292 个 mRNA 显著差异甲基化。在这些 lncRNA 中，lncRNA4114（转录 ID：ENST00000444114）在 PPDLSCs 中表现出 m6A 甲基化水平和表达水平均降低。进一步的生物信息学分析预测，差异甲基化的 mRNAs 主要参与细胞相互作用、干细胞多能性和成骨分化信号。然后，甲基转移酶样 3（METTL3）的过表达促进了 PPDLSCs 的成骨分化，而 METTL3 的敲低则表现出抑制作用。此外，METTL3 过表达促进 lncRNA4114 的稳定性，从而上调其表达水平。此外，lncRNA4114 的过表达促进了 PPDLSCs 的成骨分化。
METTL3 通过调节 lncRNA4114 的稳定性促进 PPDLSCs 的成骨分化。
