Department of Stomatology, Nanfang Hospital, Southern Medical University, Guangzhou, China; School of Stomatology, Southern Medical University, Guangzhou, China.
Stomatological Hospital, Southern Medical University, Guangzhou, China.
Arch Oral Biol. 2020 May;113:104691. doi: 10.1016/j.archoralbio.2020.104691. Epub 2020 Mar 19.
Angiogenesis is important in pulp-dentin formation. Among the regulatory factors, long noncoding RNA (LncRNA) is a class of functional RNA molecules that are not translated into protein and involved in regulating multiple physiological processes. The different expression of LncRNA and its target gene in dental pulp stem cells (DPSCs) were explored and may provide a theoretical basis for future regulation of dental pulp angiogenesis.
In this study, we cultured DPSCs from healthy dental pulp tissues and divided them into two groups: the normal DPSCs and the DPSCs cultured in vascular induction medium. In total, 40,173 LncRNA probes and 20,730 protein coding mRNAs were detected through microarray, which were then verified by the quantitative reverse transcription-polymerase chain reaction (qRT-PCR) method.
The result of differential expressions measured in LncRNA through microarray showed that 376 LncRNAs increased significantly and 426 were downregulated among the two groups of cells. Moreover, the mRNA microarray in normal cultured DPSCs showed that 629 LncRNAs were significantly upregulated, while 529 of them were downregulated compared with the DPSCs that were cultured in vascular induction medium. Gene ontology (GO) analysis inferred the molecular function of mRNAs. Pathway analysis showed that 52 signaling pathways were involved in the differentiation process of DPSCs. qRT-PCR analysis, conducted for validation, showed results consistent with the microarray analysis.
We found that a number of different regulators are involved in inducing vascular differentiation of DPSCs, which provides a foundation for subsequent experiments.
血管生成在牙髓-牙本质形成中很重要。在调节因子中，长链非编码 RNA（LncRNA）是一类不翻译成蛋白质的功能性 RNA 分子，参与调节多种生理过程。本研究通过微阵列技术探索牙髓干细胞（DPSCs）中 LncRNA 及其靶基因的不同表达，可能为未来牙髓血管生成的调控提供理论依据。
本研究培养来自健康牙髓组织的 DPSCs，并将其分为两组：正常 DPSCs 和在血管诱导培养基中培养的 DPSCs。通过微阵列共检测到 40173 个 LncRNA 探针和 20730 个蛋白质编码 mRNAs，并用定量逆转录-聚合酶链反应（qRT-PCR）方法进行验证。
通过微阵列测量的 LncRNA 差异表达结果显示，两组细胞中 376 个 LncRNA 显著上调，426 个 LncRNA 下调。此外，正常培养的 DPSCs 的 mRNA 微阵列显示，与在血管诱导培养基中培养的 DPSCs 相比，629 个 LncRNA 显著上调，而 529 个 LncRNA 下调。GO 分析推断了 mRNAs 的分子功能。通路分析显示，52 条信号通路参与了 DPSCs 的分化过程。qRT-PCR 分析验证结果与微阵列分析一致。
我们发现许多不同的调控因子参与诱导 DPSCs 的血管分化，为后续实验提供了基础。
