中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (31): 8122-8134.doi: 10.12307/2026.421

• 牙髓及牙周膜干细胞 Dental pulp and periodontal ligament stem cells • 上一篇    下一篇

人牙髓干细胞来源诱导性多能干细胞成牙本质分化的基因表达调控网络与分子机制

荣  琼，郭  宇，汪婷婷，谭小兵   

  1. 云南省第一人民医院，昆明理工大学附属医院口腔医学中心，云南省昆明市   650032
  • 出版日期:2026-11-08 发布日期:2026-05-23
  • 通讯作者: 谭小兵，硕士，主任医师，云南省第一人民医院，昆明理工大学附属医院口腔医学中心，云南省昆明市 650032
  • 作者简介:荣琼，女，1985年生，云南省曲靖市人，汉族，2013年中山大学毕业，博士，副主任医师，主要从事干细胞与牙槽骨缺损修复的研究。
  • 基金资助:
    国家自然科学基金(82160178)，项目负责人：谭小兵；国家自然科学基金(82560185)，项目负责人：荣琼

Gene expression regulatory network and molecular mechanism involved in odontogenic differentiation of human dental pulp stem cells-derived induced pluripotent stem cells

Rong Qiong, Guo Yu, Wang Tingting, Tan Xiaobing   

  1. Center of Stomatology, The First People’s Hospital of Yunnan Province, Affiliated Hospital of Kunming University of Science and Technology, Kunming 650032, Yunnan Province, China
  • Online:2026-11-08 Published:2026-05-23
  • Contact: Tan Xiaobing, MS, Chief physician, Center of Stomatology, The First People’s Hospital of Yunnan Province, Affiliated Hospital of Kunming University of Science and Technology, Kunming 650032, Yunnan Province, China
  • About author:Rong Qiong, MD, Associate chief physician, Center of Stomatology, The First People’s Hospital of Yunnan Province, Affiliated Hospital of Kunming University of Science and Technology, Kunming 650032, Yunnan Province, China
  • Supported by:
    National Natural Science Foundation of China, No. 82160178 (to TXB); National Natural Science Foundation of China, No. 82560185 (to RQ)

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摘要：

文题释义：

人牙髓干细胞来源的诱导性多能干细胞：具有来源便捷、伦理争议小、重编程效率高、良好的基因组稳定性和潜在的神经/牙源性分化等优势，成为一种极具吸引力的多能干细胞来源。它们在再生医学、疾病建模、药物筛选和个性化细胞/基因治疗等领域展现出巨大的应用潜力。随着对诱导性多能干细胞生物学特性的深入理解、分化效率改进、生物安全性和规模化生产等关键问题的逐步解决，人牙髓干细胞来源诱导性多能干细胞有望成为牙本质再生的理想种子细胞。
成牙本质分化：指干细胞在特定信号诱导下，逐步获得成牙本质细胞表型(如表达牙本质涎磷蛋白、牙本质基质蛋白1等特异性标志物)，并分泌牙本质基质(如Ⅰ型胶原等)的过程，是牙本质形成与修复再生的关键生物学事件，解析调控机制对牙体组织再生治疗具有重要意义。

摘要
背景：人牙源性诱导性多能干细胞具有自我更新和多向分化潜能，在促进牙本质再生方面具有巨大的应用前景，但其成牙本质分化的机制尚不明确。
目的：分析人牙髓干细胞来源诱导性多能干细胞成牙本质分化过程中的关键调控基因及潜在分子机制。
方法：收集人牙髓干细胞来源诱导性多能干细胞及其分化成牙本质细胞，提取总RNA建库测序，分析转录组基因表达谱。首先，筛选差异表达mRNAs、miRNAs、长链非编码RNA和环状RNA，确定关键基因后，基因集富集分析关键信号通路；然后构建预测调控关键基因的竞争性内源RNA、转录因子-mRNA-miRNA网络，揭示调控机制；最后，实时荧光定量PCR验证关键基因的表达。
结果与结论：人牙髓干细胞来源诱导性多能干细胞成牙本质分化过程共鉴定出6个关键基因：Smad家族成员3、信号传导及转录激活因子3、纤维连接蛋白1、雌激素受体1、表皮生长因子受体、Zeste同源物增强子2，最富集通路包括Wnt、转化生长因子β信号通路等。竞争性内源RNA网络分析发现hsa-miR-302、hsa-miR-519和hsa-miR-520是雌激素受体1的主要调控因子，存在多个关系对；转录因子-mRNA网络分析发现干扰素调节因子3是6个关键基因的调控因子，而Zeste同源物增强子2同时受到4个转录因子的调控。实时荧光定量PCR结果显示牙髓干细胞来源诱导性多能干细胞分化的成牙本质细胞中纤维连接蛋白1和Zeste同源物增强子2的表达显著升高，而信号传导及转录激活因子3的表达显著降低。

关键词: 人牙髓干细胞, 诱导性多能干细胞, 成牙本质分化, 转录组测序, 关键基因, 差异表达基因, 基因调控网络, 竞争性内源RNA网络

Abstract: BACKGROUND: Human dental pulp stem cell-derived induced pluripotent stem cells possess self-renewal and multidirectional differentiation potential, showing great promise in promoting dentin regeneration. However, the mechanism of their odontogenic differentiation remains unclear.
OBJECTIVE: To analyze the hub genes and potential molecular mechanisms during the odontogenic differentiation of human dental pulp stem cell-derived induced pluripotent stem cells.
METHODS: Total RNA of human dental pulp stem cell-derived induced pluripotent stem cells and their differentiated odontogenic cells were extracted for RNA-sequencing to analyze the transcriptome gene expression profile. Firstly, the differentially expressed mRNAs, miRNAs, long non-coding RNAs, and circular RNAs were screened to identify key genes. Gene Set Enrichment Analysis was performed to analyze the key signaling pathways. Subsequently, a competitive endogenous RNA (ceRNA) network and a transcription factor-mRNA-miRNA network were constructed to predict and reveal the regulatory mechanisms of key genes. Finally, real-time quantitative PCR was used to verify the expression of key genes. 
RESULTS AND CONCLUSION: Six hub genes were identified during the odontogenic differentiation of human dental pulp stem cell-derived induced pluripotent stem cells: Smad family member 3, signal transducer and activator of transcription 3, fibronectin 1, estrogen receptor 1, epidermal growth factor receptor, and enhancer of zeste homolog 2. The most enriched pathways included Wnt and transforming growth factor β signaling pathways. CeRNA network analysis revealed that hsa-miR-302, hsa-miR-519, and hsa-miR-520 were the main regulatory factors of estrogen receptor 1, with multiple interaction pairs. Transcription factor-mRNA network analysis showed that interferon regulatory factor 3 was a regulatory factor for all six hub genes, while enhancer of zeste homolog 2 was regulated by four transcription factors simultaneously. Real-time quantitative PCR results showed that the expression of fibronectin 1 and enhancer of zeste homolog 2 was significantly increased in odontogenic cells differentiated from dental pulp stem cell-derived induced pluripotent stem cells, while the expression of signal transducer and activator of transcription 3 was significantly decreased.


Key words: human dental pulp stem cells, induced pluripotent stem cells, odontogenic differentiation, RNA sequencing, key genes, differentially expressed genes, gene regulatory network, competitive endogenous RNA network

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