Tan Dan, Li Qilin, Chen Zhenzhen, Zhang Hongbing, Rao Pengcheng, Li Jingxiang, Tao Qianke, Xiao Jingang, Song Jinlin
Chongqing Key Laboratory of Oral Diseases and Biomedical Sciences, Chongqing Municipal Key Laboratory of Oral Biomedical Engineering of Higher Education, College of Stomatology of Chongqing Medical University, Chongqing, China.
Luzhou Key Laboratory of Oral & Maxillofacial Reconstruction and Regeneration, the Affiliated Stomatological Hospital, Southwest Medical University, Luzhou, China.
Cell Prolif. 2025 Aug;58(8):e70020. doi: 10.1111/cpr.70020. Epub 2025 Mar 17.
Human periodontal ligament stem cells (hPDLSCs) have emerged as promising candidates for the treatment of osteoporotic bone defects. Previous studies have indicated that mA plays a crucial role in regulating the osteogenic differentiation of hPDLSCs. However, research on the relationship between YTHDC1, as a reading protein, and the osteogenic differentiation of hPDLSCs remains unexplored. This study aimed to investigate the biological roles of YTHDC1 in the osteogenic differentiation of hPDLSCs and to explore underlying mechanisms. Dot blot analysis revealed a progressive increase in mA methylation during osteogenic differentiation, accompanied by significant upregulation of YTHDC1 expression, as evidenced by qPCR and Western blot. Functional assays utilising siRNA-mediated knockdown and lentiviral-mediated overexpression demonstrated that YTHDC1 positively regulated the osteogenic differentiation potential of hPDLSCs. Mechanistically, mRNA-seq analysis implicated the Wnt/β-catenin signalling pathway, which was further validated through rescue experiments with the Wnt inhibitor DKK1. Notably, in vivo experiments showed that hPDLSCs overexpressing YTHDC1 exhibited enhanced bone formation capacity in the osteoporotic rats. In conclusion, our findings suggested that YTHDC1 modulated the osteogenic capacity of hPDLSCs through the Wnt/β-catenin signalling pathway, highlighting its therapeutic potential for treating bone defects in osteoporotic conditions.
人牙周膜干细胞（hPDLSCs）已成为治疗骨质疏松性骨缺损的有前景的候选细胞。先前的研究表明，m⁶A在调节hPDLSCs的成骨分化中起关键作用。然而，作为一种读取蛋白的YTHDC1与hPDLSCs成骨分化之间的关系研究仍未开展。本研究旨在探讨YTHDC1在hPDLSCs成骨分化中的生物学作用，并探索其潜在机制。斑点杂交分析显示，在成骨分化过程中m⁶A甲基化逐渐增加，同时qPCR和蛋白质印迹证明YTHDC1表达显著上调。利用小干扰RNA介导的敲低和慢病毒介导的过表达进行的功能试验表明，YTHDC1正向调节hPDLSCs的成骨分化潜能。机制上，mRNA测序分析涉及Wnt/β-连环蛋白信号通路，通过用Wnt抑制剂DKK1进行的拯救实验进一步验证了该通路。值得注意的是，体内实验表明，过表达YTHDC1的hPDLSCs在骨质疏松大鼠中表现出增强的骨形成能力。总之，我们的研究结果表明，YTHDC1通过Wnt/β-连环蛋白信号通路调节hPDLSCs的成骨能力，突出了其在治疗骨质疏松症骨缺损方面的治疗潜力。
