Nanfang Hospital, Southern Medical University, Guangzhou, People's Republic of China.
School of Stomatology, Southern Medical University, Guangzhou, People's Republic of China.
Stem Cell Res Ther. 2022 Jun 28;13(1):286. doi: 10.1186/s13287-022-02970-5.
Human dental pulp stem cells (hDPSCs) are critical for pulp generation. hDPSCs proliferate faster under hypoxia, but the mechanism by which long noncoding RNA (lncRNA) regulates this process is not fully understood.
Novel lncRNAs were obtained by reanalysis of transcriptome datasets from RNA-Seq under hypoxia compared with normoxia, and a differential expression analysis of target genes was performed. Bioinformatics analyses, including gene ontology analysis, Kyoto Encyclopedia of Genes and Genomes pathway analysis and gene set enrichment analysis, were used to understand the function of key novel lncRNAs. hDPSCs were isolated from dental pulp tissue. EdU and scratch wound healing assays were used to detect the proliferation and migration of hDPSCs. qRT-PCR was used to detect changes in the RNA expression of selected genes. RNA fluorescence in situ hybridization, small interfering RNA, qRT-PCR and Western blot analysis were used to explore the function of key novel lncRNAs.
We identified 496 novel lncRNAs in hDPSCs under hypoxia, including 45 differentially expressed novel lncRNAs. Of these, we focused on a key novel lncRNA, which we designated HRL-SC (hypoxia-responsive lncRNA in stem cells). Functional annotation revealed that HRL-SC was associated with hypoxic conditions and the PI3K/AKT signaling pathway. HRL-SC was mainly located in the cytoplasm of hDPSCs and had stable high expression under hypoxia. Knockdown of HRL-SC inhibited the proliferation and migration of hDPSCs and the expression levels of PI3K/AKT-related marker proteins. Furthermore, the AKT activator SC79 partially offset the inhibitory effect caused by the knockdown, indicating that HRL-SC promoted hDPSCs through the PI3K/AKT signaling pathway.
Hypoxia-responsive lncRNA HRL-SC promotes the proliferation and migration of hDPSCs through the PI3K/AKT signaling pathway, and this understanding may facilitate the regenerative application of hDPSCs.
人牙髓干细胞（hDPSCs）对于牙髓再生至关重要。hDPSCs 在低氧环境下增殖更快，但长链非编码 RNA（lncRNA）调控这一过程的机制尚未完全阐明。
通过对低氧与常氧条件下 RNA-Seq 转录组数据集的重新分析，获得新的 lncRNA，并对靶基因进行差异表达分析。采用基因本体分析、京都基因与基因组百科全书通路分析和基因集富集分析等生物信息学分析，以了解关键新 lncRNA 的功能。从牙髓组织中分离 hDPSCs。EdU 和划痕愈合实验用于检测 hDPSCs 的增殖和迁移。qRT-PCR 用于检测选定基因的 RNA 表达变化。RNA 荧光原位杂交、小干扰 RNA、qRT-PCR 和 Western blot 分析用于探索关键新 lncRNA 的功能。
我们在 hDPSCs 低氧条件下鉴定出 496 个新的 lncRNA，包括 45 个差异表达的新 lncRNA。其中，我们重点关注一个关键的新 lncRNA，我们将其命名为 HRL-SC（干细胞中对低氧反应的 lncRNA）。功能注释显示，HRL-SC 与低氧条件和 PI3K/AKT 信号通路有关。HRL-SC 主要位于 hDPSCs 的细胞质中，在低氧条件下表达稳定且较高。敲低 HRL-SC 抑制 hDPSCs 的增殖和迁移，以及 PI3K/AKT 相关标记蛋白的表达水平。此外，AKT 激活剂 SC79 部分抵消了敲低引起的抑制作用，表明 HRL-SC 通过 PI3K/AKT 信号通路促进 hDPSCs。
低氧反应性 lncRNA HRL-SC 通过 PI3K/AKT 信号通路促进 hDPSCs 的增殖和迁移，这一发现可能有助于 hDPSCs 的再生应用。
