Department Prosthodontics, Peking University School and Hospital of Stomatology, National Engineering Laboratory for Digital and Material Technology of Stomatology, Beijing Key Laboratory of Digital Stomatology, PR China.
Department Prosthodontics, Peking University School and Hospital of Stomatology, National Engineering Laboratory for Digital and Material Technology of Stomatology, Beijing Key Laboratory of Digital Stomatology, PR China.
Cell Signal. 2018 Dec;52:65-73. doi: 10.1016/j.cellsig.2018.08.015. Epub 2018 Aug 27.
Long noncoding RNAs (lncRNAs) are emerging as important regulators in molecular processes and may play vital roles in odontogenic differentiation of human dental pulp stem cells (hDPSCs). However, their functions remain to be elucidated. As lncRNA H19 is one of the most classical lncRNA, which plays essential roles in cellular differentiation, thus we explored the effects and mechanisms of H19 in odontogenic differentiation of hDPSCs. Stable overexpression and knockdown of H19 in hDPSCs were constructed using recombinant lentiviruses containing H19 and short hairpin-H19 expression cassettes, respectively. Alkaline phosphatase (ALP) assay, Alizarin red staining assay, von kossa staining, quantitative polymerase chain reaction (qPCR), Western blot analysis, and immunofluorescent staining results indicated that overexpression of H19 in hDPSCs positively regulates the odontogenic differentiation of hDPSCs, while knockdown of H19 in hDPSCs inhibits odontogenic differentiation of hDPSCs. Further, we found that H19 promotes the odontogenic differentiation of hDPSCs through S-adenosylhomocysteine hydrolase (SAHH) epigenetically regulates the methylation and expression of distal-less homeobox (DLX3) gene. Herein, for the first time, we determined that H19/SAHH axis epigentically regulates odontogenic differentiaiton of hDPSCs by inhibiting the DNA methyltransferase 3B (DNMT3B)-mediated methylation of DLX3. Our findings provide a new insight into how H19/SAHH axis play its role in odontogenic differentiation of hDPSCs, and would be helpful in developing therapeutic approaches for dentin regeneration based on stem cells.
长链非编码 RNA（lncRNA）作为分子过程中的重要调控因子而出现，它们可能在人牙髓干细胞（hDPSCs）的牙源性分化中发挥重要作用。然而，其功能仍有待阐明。由于 lncRNA H19 是最经典的 lncRNA 之一，在细胞分化中起着至关重要的作用，因此我们探讨了 H19 在 hDPSCs 牙源性分化中的作用及其机制。使用含有 H19 和短发夹-H19 表达盒的重组慢病毒分别构建 hDPSCs 中 H19 的稳定过表达和敲低载体。碱性磷酸酶（ALP）检测、茜素红染色检测、von kossa 染色、实时定量聚合酶链反应（qPCR）、Western blot 分析和免疫荧光染色结果表明，hDPSCs 中 H19 的过表达正向调节 hDPSCs 的牙源性分化，而 hDPSCs 中 H19 的敲低抑制 hDPSCs 的牙源性分化。此外，我们发现 H19 通过 S-腺苷同型半胱氨酸水解酶（SAHH）表观遗传调控远端同源盒（DLX3）基因的甲基化和表达来促进 hDPSCs 的牙源性分化。在此，我们首次确定 H19/SAHH 轴通过抑制 DNA 甲基转移酶 3B（DNMT3B）介导的 DLX3 甲基化来表观遗传调控 hDPSCs 的牙源性分化。我们的研究结果为 H19/SAHH 轴在 hDPSCs 牙源性分化中的作用提供了新的见解，并有助于开发基于干细胞的牙本质再生治疗方法。
