Understanding the regulation mechanisms of mesenchymal stem cells (MSCs) can assist in tissue regeneration. The histone demethylase (KDM) family has a crucial role in differentiation and cell proliferation of MSCs, while the function of KDM3B in MSCs is not well understood. In this study, we used the stem cells from the apical papilla (SCAPs) to test whether KDM3B could regulate the function of MSCs. By an alkaline phosphatase (ALP) activity assay, Alizarin red staining, real-time RT-PCR, and western blot analysis, we found that KDM3B enhanced the ALP activity and mineralization of SCAPs and promoted the expression of runt-related transcription factor 2 (RUNX2), osterix (OSX), dentin sialophosphoprotein (DSPP), and osteocalcin (OCN). Additionally, the CFSE, CCK-8, and flow cytometry assays revealed that KDM3B improved cell proliferation by accelerating cell cycle transition from the G1 to S phase. Scratch and transwell migration assays displayed that KDM3B promoted the migration potential of SCAPs. Mechanically, microarray results displayed that 98 genes were upregulated, including STAT1, CCND1, and FGF5, and 48 genes were downregulated after KDM3B overexpression. Besides, we found that the Toll-like receptor and JAK-STAT signaling pathway may be involved in the regulating function of KDM3B in SCAPs. In brief, we discovered that KDM3B promoted the osteo-/odontogenic differentiation, cell proliferation, and migration potential of SCAPs and provided a novel target and theoretical basis for regenerative medicine.

中文翻译：

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组蛋白去甲基化酶KDM3B促进根尖乳头干细胞的成骨/成牙细胞分化，细胞增殖和迁移潜能。

了解间充质干细胞（MSCs）的调节机制可以帮助组织再生。组蛋白脱甲基酶（KDM）家族在MSC的分化和细胞增殖中起着至关重要的作用，而人们对MSC中KDM3B的功能还知之甚少。在这项研究中，我们使用了根尖乳头（SCAPs）的干细胞来测试KDM3B是否可以调节MSC的功能。通过碱性磷酸酶（ALP）活性测定，茜素红染色，实时RT-PCR和Western blot分析，我们发现KDM3B增强了ALP活性和SCAP的矿化作用，并促进了矮子相关转录因子2（ RUNX2），osterix（OSX），牙本质唾液磷蛋白（DSPP）和骨钙蛋白（OCN）。此外，CFSE，CCK-8，流式细胞仪检测和流式细胞仪检测表明，KDM3B通过加速细胞周期从G1到S期的转变来改善细胞增殖。刮擦和穿井迁移分析表明，KDM3B促进了SCAP的迁移潜力。机械上，微阵列结果显示98个基因被上调，包括STAT1，CCND1和FGF5以及48个基因在KDM3B过表达后被下调。此外，我们发现Toll样受体和JAK-STAT信号通路可能参与了SCAPs中KDM3B的调控功能。简而言之，我们发现KDM3B促进了SCAPs的成骨/成牙细胞分化，细胞增殖和迁移潜力，并为再生医学提供了新的靶标和理论基础。