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首页> 外文期刊>Cell death & disease. >LncRNA H19 promotes the committed differentiation of stem cells from apical papilla via miR-141/SPAG9 pathway
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LncRNA H19 promotes the committed differentiation of stem cells from apical papilla via miR-141/SPAG9 pathway

机译:LncRNA H19通过miR-141 / SPAG9途径促进干细胞从根尖乳头的定向分化

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摘要

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.
机译:长非编码RNA(lncRNA)在转录和转录后水平上发挥重要作用。在某些条件下,来自根尖乳头(SCAP)的干细胞分化为牙本质/骨样组织。到目前为止,lncRNA-H19是否可以影响SCAPs的增殖行为和成骨/成牙作用及其具体机制还有待阐明。在这里,分离出SCAP并用慢病毒或包装载体转染。我们的结果表明,如CCK-8分析,EdU分析和流式细胞仪(FCM)所述,lncRNA-H19对SCAP的增殖行为没有显着影响。此外,进行了碱性磷酸酶(ALP)活性,茜素红染色,蛋白质印迹测定(WB),定量实时聚合酶链反应(qRT-PCR)和体内骨形成测定,以验证H19对SCAP的生物学影响。 H19的过表达导致SCAP的骨/牙生成增强,而敲低H19则抑制了这些作用。从机理上讲,H19与miR-141竞争结合并阻止SPAG9受到miRNA介导的降解,从而显着提高p38和JNK的磷酸化水平,并促进SCAP的持续分化。总之,通过miR-141 / SPAG9途径的H19过表达,SCAPs的骨/牙生成被上调。

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