Introduction. Odontogenic differentiation of human dental pulp stem cells (hDPSCs) is a key step of pulp regeneration. Recent studies showed that circular RNAs (circRNAs) have many biological functions and that competing endogenous RNA (ceRNA) is their most common mechanism of action. However, the role of circRNAs in hDPSCs during odontogenesis is still unclear. Methods. Isolated hDPSCs were cultured in essential and odontogenic medium. Total RNA was extracted after 14 days of culture, and then, microarray analysis was performed to measure the differential expressions of circRNAs. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was then performed to validate the microarray results. Based on microarray data from this study and available in the database, a ceRNA network was constructed to investigate the potential function of circRNAs during odontogenesis. In addition, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed to investigate the potential correlation between signaling pathways and circRNAs. In addition, qRT-PCR and Western blot analysis were used to explore the function of hsa_circRNA_104101. Results. We found 43 upregulated circRNAs and 144 downregulated circRNAs during the odontogenic differentiation process ( and <-1.5, respectively; ). qRT-PCR results were in agreement with the microarray results. Bioinformatic analysis revealed that the Wnt signaling pathway and the TGF-β signaling pathway, as well as the other pathways associated with odontogenic differentiation, were correlated to the differentially expressed circRNAs. hsa_circRNA_104101 was proved to promote the odontogenic differentiation of hDPSCs. Conclusion. This study reported 187 circRNAs that were differentially expressed in hDPSCs during odontogenic differentiation. Bioinformatic analysis of the expression data suggested that circRNA-miRNA-mRNA networks might act as a crucial mechanism for hDPSC odontogenic differentiation, providing a theoretical foundation for the study of pulp regeneration regulation by circRNAs.

中文翻译：

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牙源性分化过程中牙髓干细胞中的 circRNA 表达谱。

简介。人牙髓干细胞（hDPSCs）的成牙本质分化是牙髓再生的关键步骤。最近的研究表明，环状 RNA (circRNA) 具有许多生物学功能，竞争性内源性 RNA (ceRNA) 是它们最常见的作用机制。然而，环状RNA在牙发育过程中hDPSCs中的作用仍不清楚。方法. 分离的 hDPSCs 在基本和牙源性培养基中培养。培养14天后提取总RNA，然后进行微阵列分析以测量circRNA的差异表达。然后进行定量逆转录聚合酶链反应（qRT-PCR）以验证微阵列结果。基于该研究的微阵列数据和数据库中可用的数据，构建了一个 ceRNA 网络来研究 circRNA 在牙发育过程中的潜在功能。此外，还进行了基因本体论 (GO) 和京都基因和基因组百科全书 (KEGG) 分析，以研究信号通路与 circRNA 之间的潜在相关性。此外，qRT-PCR和Western印迹分析用于探索hsa_circRNA_104101的功能。结果. 我们在牙源性分化过程中发现了 43 个上调的 circRNA 和 144 个下调的 circRNA。和 <-1.5，分别；）。qRT-PCR 结果与微阵列结果一致。生物信息学分析表明，Wnt 信号通路和 TGF- β信号通路以及其他与牙源性分化相关的通路与差异表达的 circRNA 相关。hsa_circRNA_104101 被证明可以促进 hDPSCs 的牙源性分化。结论。该研究报告了 187 个在牙源性分化过程中在 hDPSCs 中差异表达的 circRNA。表达数据的生物信息学分析表明，circRNA-miRNA-mRNA网络可能是hDPSC牙源性分化的关键机制，为circRNAs对牙髓再生调控的研究提供了理论基础。