The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) and Key Laboratory of Oral Biomedicine Ministry of Education, School and Hospital of Stomatology, Wuhan University, Wuhan 430079, China.
Department of Prosthodontics, Yinchuan Stomatology Hospital, Yinchuan 750002, China.
Tissue Cell. 2023 Jun;82:102081. doi: 10.1016/j.tice.2023.102081. Epub 2023 Mar 26.
The N6-methyladenosine (m6A) modification has been proven to be involved in various physiological and pathological processes. The m6A is catalyzed by methyltransferase complex, which mainly consist of methyltransferase (METTL) 3 and 14 heterodimer. The present study aimed to investigate the role of METTL 3 and 14 in biological properties of periodontal ligament cells (PDLCs) via RNA-sequencing and specific cell assays. Firstly, the expressions of METTL3 and METTL14 were observed in PDLCs. Then, RNA-sequencing showed that cell properties were influenced after METTL3 or METTL14 was knocked down via short hairpin RNA (shRNA). In sh-METTL3 or METTL14 PDLCs, cell counting kit 8 (CCK8) and 5-ethynyl-2'-deoxyuridine (EdU) assays showed a down-regulated proliferation, transwell system indicated suppressed migration. Lastly, alkaline phosphatase (ALP) and alizarin red staining (ARS) staining, quantitative polymerase chain reaction (qPCR) and western blot demonstrated the inhibited osteogenic potentials. It could be concluded that METTL3 and METTL14 play indispensable roles in the regenerative potential of PDLCs.
N6-甲基腺苷（m6A）修饰已被证明参与多种生理和病理过程。m6A 由甲基转移酶复合物催化，主要由甲基转移酶（METTL）3 和 14 异二聚体组成。本研究旨在通过 RNA 测序和特定的细胞实验，研究 METTL3 和 14 在牙周膜细胞（PDLCs）生物学特性中的作用。首先，观察 PDLCs 中 METTL3 和 METTL14 的表达。然后，RNA 测序显示，通过短发夹 RNA（shRNA）敲低 METTL3 或 METTL14 后，细胞特性受到影响。在 sh-METTL3 或 METTL14 PDLCs 中，细胞计数试剂盒 8（CCK8）和 5-乙炔基-2'-脱氧尿苷（EdU）检测显示增殖下调，Transwell 系统表明迁移受到抑制。最后，碱性磷酸酶（ALP）和茜素红染色（ARS）染色、定量聚合酶链反应（qPCR）和蛋白质印迹实验表明成骨潜能受到抑制。综上所述，METTL3 和 METTL14 在 PDLCs 的再生潜能中发挥不可或缺的作用。
