Department of Operative Dentistry and Endodontics, Guanghua School of Stomatology, Affiliated Stomatological Hospital, Guangdong Province Key Laboratory of Stomatology, Sun Yat‑sen University, Guangzhou, Guangdong 510055, P.R. China.
Mol Med Rep. 2019 Aug;20(2):871-878. doi: 10.3892/mmr.2019.10359. Epub 2019 Jun 6.
The alterations in expression and function of circular RNA (circRNA) in human dental pulp cells (hDPCs) during odontogenic differentiation were investigated. To induce odontogenic differentiation, hDPCs (passage 3) were cultured for 14 days in odontogenic induction medium. circRNA high‑throughput sequencing was performed using Illumina HiSeqseq™ 2000. Subsequently, Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses were used to evaluate the bio‑functions of the identified circRNAs. To validate the results of circRNA sequencing, reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) was performed for two selected differentially expressed circRNAs. The RNA sequencing results revealed that 1,314 and 1,780 circRNAs were upregulated and downregulated, respectively, during odontogenic induction. Their predicted target miRNAs and genes are involved in several biological functions and signaling pathways, including the mitogen‑associated protein kinase signaling pathway. The RT‑qPCR results of the two selected circRNAs (hsa_circ_0015260 and hsa_circ_0006984) were consistent with the expression trend obtained using high‑throughput sequencing. The results of the present study add to the current understanding of the regulatory mechanisms underlying hDPCs differentiation.
研究了人牙髓细胞（hDPC）在牙源性分化过程中环状 RNA（circRNA）表达和功能的改变。为了诱导牙源性分化，将 hDPC（第 3 代）在牙源性诱导培养基中培养 14 天。使用 Illumina HiSeqseq™2000 进行 circRNA 高通量测序。随后，使用基因本体论和京都基因与基因组百科全书分析来评估鉴定出的 circRNAs 的生物功能。为了验证 circRNA 测序的结果，对两个选定的差异表达 circRNA 进行了逆转录-定量聚合酶链反应（RT-qPCR）。RNA 测序结果显示，在牙源性诱导过程中，分别有 1314 个和 1780 个 circRNA 上调和下调。它们预测的靶 miRNA 和基因参与了几个生物学功能和信号通路，包括丝裂原激活蛋白激酶信号通路。使用高通量测序获得的两个选定 circRNA（hsa_circ_0015260 和 hsa_circ_0006984）的 RT-qPCR 结果与表达趋势一致。本研究的结果增加了对 hDPC 分化调控机制的理解。
