Department of Stomatology, Nanfang Hospital, Southern Medical University, Guangzhou, PR China; College of Stomatology, Southern Medical University, Guangzhou, PR China.
College of Stomatology, Southern Medical University, Guangzhou, PR China; Department of Operative and Endodontic Dentistry, Stomatological Hospital, Southern Medical University, Guangzhou, PR China.
J Proteomics. 2019 Jun 30;202:103364. doi: 10.1016/j.jprot.2019.04.014. Epub 2019 Apr 19.
Odontoblastic/osteogenic differentiation of human dental pulp stem cells (hDPSCs) is a key factor in tooth and pulp regeneration, but its mechanism still remains unknown. The purpose of this research is to look into the mechanism by which Stathmin affects the proliferation and odontoblastic/osteogenic differentiation of hDPSCs, and whether the Wnt/β- catenin is related to this regulation. First, the Stathmin expression was inhibited by lentiviral vector, after that the transcriptome sequencing technology was used to screen the differentially expressed genes, then we found Wnt5a which related to the regulation of Wnt/β-catenin was regulated. Comparing with hDPSC in the control group, the shRNA-Stathmin group inhibited proliferation and odontoblastic/osteogenic differentiation. The result of molecular analysis indicated that the Wnt/β-catenin was inhibited when Stathmin was silenced. After that, the shRNA-Stathmin group were added with LiCl (activator of Wnt/β-catenin), and the Wnt/β-catenin was significantly activated in β-catenin. After activation of the Wnt/β-catenin, the proliferation of hDPSCs was significantly increased and the expression of genes related to odontoblastic/osteogenic differentiation was also significantly increased. Taken together, these findings reveal for the first time that the Stathmin-Wnt/β-catenin plays a positive regulatory role in hDPSC proliferation and odontoblastic/osteogenic differentiation. SIGNIFICANCE: Transcriptome sequencing revealed that Stathmin interacts with Wnt/β-catenin signaling pathway-related proteins such as Wnt5a. At the same time, experiments have confirmed that Stathmin protein can affect the proliferation and odontogenetic differentiation of hDPSCs.The innovation of this paper is to link the Stathmin and Wnt/β-catenin signaling pathways for the first time, to explore the interaction of Stathmin and Wnt/β-catenin signaling pathways and the mechanism of this regulation on human dental pulp stem cells (hDPSCs) of odontoblastic/osteogenic differentiation and proliferation function. Especially for the regulation of odontoblastic/osteogenic differentiation, we have verified this mechanism at the molecular level and characterization leveland this regulation also provides new ideas for dental pulp tissue engineering. At the same time, more than 3000 proteins related to the change of Stathmin level were screened by transcriptome sequencing technology, which provided a possibility to further exploration of the regulation mechanism of Stathmin on various aspects of cell biological characteristics.
牙髓干细胞（hDPSCs）的成牙本质/成骨分化是牙齿和牙髓再生的关键因素，但其机制尚不清楚。本研究旨在探讨 Stathmin 影响 hDPSCs 增殖和牙本质/成骨分化的机制，以及 Wnt/β-连环蛋白是否与这种调节有关。首先，通过慢病毒载体抑制 Stathmin 的表达，然后使用转录组测序技术筛选差异表达基因，发现与 Wnt/β-catenin 调节相关的 Wnt5a 受到调节。与对照组 hDPSC 相比，shRNA-Stathmin 组抑制增殖和牙本质/成骨分化。分子分析结果表明，沉默 Stathmin 时 Wnt/β-catenin 受到抑制。之后，向 shRNA-Stathmin 组中加入 LiCl（Wnt/β-catenin 的激活剂），β-连环蛋白中的 Wnt/β-catenin 明显被激活。Wnt/β-catenin 激活后，hDPSCs 的增殖明显增加，牙本质/成骨分化相关基因的表达也明显增加。综上所述，这些发现首次揭示了 Stathmin-Wnt/β-catenin 正向调控 hDPSC 增殖和牙本质/成骨分化。意义：转录组测序显示，Stathmin 与 Wnt/β-连环蛋白信号通路相关蛋白如 Wnt5a 相互作用。同时，实验证实 Stathmin 蛋白可影响 hDPSCs 的增殖和牙发生分化。本文的创新之处在于首次将 Stathmin 与 Wnt/β-catenin 信号通路联系起来，探索 Stathmin 与 Wnt/β-catenin 信号通路的相互作用及其对人牙髓干细胞（hDPSCs）牙本质/成骨分化和增殖功能的调节机制。特别是对于牙本质/成骨分化的调节，我们已经在分子水平和表征水平上验证了这一机制，这一调节也为牙髓组织工程提供了新的思路。同时，通过转录组测序技术筛选出与 Stathmin 水平变化相关的 3000 多个蛋白，为进一步探讨 Stathmin 对细胞生物学特性各方面的调节机制提供了可能。
