LncRNA H19 promotes the committed differentiation of stem cells from apical papilla via miR-141/SPAG9 pathway
LncRNA H19 promotes the committed differentiation of stem cells from apical papilla via miR-141/SPAG9 pathway
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LncRNA H19通过miR-141/SPAG9通路促进根尖乳头干细胞定向分化
DOI：
10.1038/s41419-019-1337-3
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发表时间：
2019-02-12
影响因子：
9
通讯作者：
Yu, Jinhua
中科院分区：
文献类型：
--
作者：
Li, Zehan;Yan, Ming;Yu, Jinhua
长链非编码RNA（lncRNA）在转录和转录后水平发挥重要作用。根尖乳头干细胞（Stem cells from apical papilla，SCAPs）在一定条件下可分化为牙本质/骨样组织。目前，lncRNA-H19是否能影响SCAPs的增殖行为和成骨成牙，其具体机制尚不清楚。在此，分离SCAPs并用慢病毒或包装载体转染。CCK-8法、EdU法和流式细胞仪检测结果显示，lncRNA-H19对SCAPs的增殖行为无明显影响。通过碱性磷酸酶（ALP）活性测定、茜素红染色、Western blot、实时荧光定量PCR和体内成骨实验，验证H19对SCAPs的生物学影响。H19的过表达导致SCAPs的骨/牙生成增强，而H19的敲低抑制了这些作用。在机制上，H19竞争性结合miR-141并阻止SPAG 9由miRNA介导的降解，从而显著提高p38和JNK的磷酸化水平并促进SCAPs的定向分化。总之，通过miR-141/SPAG 9途径过表达H19上调SCAPs的骨/牙发生。
Long noncoding RNAs (lncRNAs) exert significant roles at transcriptional and post-transcriptional levels. Stem cells from apical papilla (SCAPs) differentiate into dentin/bone-like tissues under certain conditions. So far, whether lncRNA-H19 can affect the proliferative behaviors and osteo/odontogenesis of SCAPs, as well as its specific mechanism remain to be elucidated. Here, SCAPs were isolated and transfected with the lentiviruses or packaging vectors. Our results showed that lncRNA-H19 had no significant effect on the proliferative behaviors of SCAPs, as presented by CCK-8 assay, EdU assay and flow cytometry (FCM). Furthermore, alkaline phosphatase (ALP) activity, alizarin red staining, Western blot assay (WB), quantitative real-time polymerase chain reaction (qRT-PCR) and in vivo bone formation assay were conducted to verify the biological influences of H19 on SCAPs. Overexpression of H19 led to the enhanced osteo/odontogenesis of SCAPs, whereas knockdown of H19 inhibited these effects. Mechanistically, H19 competitively bound to miR-141 and prevented SPAG9 from miRNA-mediated degradation, thus significantly elevating phosphorylated levels of p38 and JNK and facilitating the committed differentiation of SCAPs. Taken together, the osteo/odontogenesis of SCAPs was upregulated by overexpression of H19 via miR-141/SPAG9 pathway.