Long non-coding RNA H19/SAHH axis epigenetically regulates odontogenic differentiation of human dental pulp stem cells
Long non-coding RNA H19/SAHH axis epigenetically regulates odontogenic differentiation of human dental pulp stem cells
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长非编码RNA H19/SAHH轴表观遗传调控人牙髓干细胞的牙源性分化
DOI：
10.1016/j.cellsig.2018.08.015
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发表时间：
2018
影响因子：
4.8
通讯作者：
Wang Yixiang
中科院分区：
文献类型：
--
作者：
Zeng Li;Sun Shichen;Han Dong;Liu Yang;Liu Haochen;Feng Hailan;Wang Yixiang
长链非编码rna （lncRNAs）作为重要的分子调控因子在人牙髓干细胞（hDPSCs）的成牙分化过程中发挥着重要作用。然而，它们的功能仍有待阐明。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，因此我们探索了H19在hDPSCs成牙分化中的作用及其机制。利用含有H19和短发夹H19表达带的重组慢病毒，分别构建了H19在hDPSCs中的稳定过表达和低表达。碱性磷酸酶（ALP）、茜素红染色、von kossa染色、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过表达可正向调节hDPSCs的成牙分化，而H19过表达可抑制hDPSCs的成牙分化。此外，我们发现H19通过s -腺苷型同型半胱氨酸水解酶（SAHH）通过表观遗传调控远端同源盒（DLX3）基因的甲基化和表达，促进hdpsc的成牙性分化。在这里，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，从表观遗传学上调控hdpsc的成牙性分化。我们的研究结果为H19/SAHH轴如何在hdpsc的牙源性分化中发挥作用提供了新的见解，并将有助于开发基于干细胞的牙本质再生治疗方法。
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.