Wang Jing-Jing, Dong Rui, Wang Li-Ping, Wang Jin-Song, Du Juan, Wang Song-Lin, Shan Zhao-Chen, Fan Zhi-Peng
Laboratory of Molecular Signaling and Stem Cells Therapy, Beijing Key Laboratory of Tooth Regeneration and Function Reconstruction, Capital Medical University School of Stomatology Beijing 100050, China ; Outpatient Department of Oral and Maxillofacial Surgery, Capital Medical University School of Stomatology Beijing 100050, China.
Laboratory of Molecular Signaling and Stem Cells Therapy, Beijing Key Laboratory of Tooth Regeneration and Function Reconstruction, Capital Medical University School of Stomatology Beijing 100050, China.
Int J Clin Exp Med. 2015 Feb 15;8(2):2165-73. eCollection 2015.
Mesenchymal stem cells (MSCs) are a reliable resource for tissue regeneration, but the molecular mechanism underlying directed differentiation remains unclear; this has restricted potential MSC applications. Histone methylation, controlled by histone methyltransferases and demethylases, may play a key role in MSCs differentiation. Previous studies determined that KDM2B can regulate the cell proliferation and osteo/dentinogenic differentiation of MSCs. It is not known whether KDM2B is involved in the other cell lineages differentiation of MSCs. Here we used the stem cells from apical papilla (SCAPs) to study the role of KDM2B on the chondrogenic differentiation potentials in MSCs. In this study, Gain- and loss-of-function assays were applied to investigate the role of KDM2B on the chondrogenic differentiation. Alcian Blue Staining and Quantitative Analysis were used to investigate the synthesis of proteoglycans by chondrocytes. Real-time RT-PCR was used to detect the expressions of chondrogenesis related genes. The Alcian Blue staining and Quantitative Analysis results revealed that overexpression of KDM2B decreased the proteoglycans production, and real-time RT-PCR results showed that the expressions of the chondrogenic differentiation markers, COL1, COL2 and SOX9 were inhibited by overexpression of KDM2B in SCAPs. On the contrary, depletion of KDM2B increased the proteoglycans production, and inhibited the expressions of COL1, COL2 and SOX9. In conclusion, our results indicated that KDM2B is a negative regulator of chondrogenic differentiation in SCAPs and suggest that inhibition of KDM2B might improve MSC mediated cartilage regeneration.
间充质干细胞（MSCs）是组织再生的可靠来源，但定向分化背后的分子机制仍不清楚；这限制了MSCs的潜在应用。由组蛋白甲基转移酶和去甲基酶控制的组蛋白甲基化可能在MSCs分化中起关键作用。先前的研究确定KDM2B可以调节MSCs的细胞增殖和骨/牙本质分化。尚不清楚KDM2B是否参与MSCs的其他细胞谱系分化。在这里，我们使用根尖乳头干细胞（SCAPs）来研究KDM2B在MSCs软骨分化潜能中的作用。在本研究中，应用功能获得和功能丧失试验来研究KDM2B在软骨分化中的作用。采用阿尔新蓝染色和定量分析来研究软骨细胞蛋白聚糖的合成。实时RT-PCR用于检测软骨生成相关基因的表达。阿尔新蓝染色和定量分析结果显示，KDM2B的过表达降低了蛋白聚糖的产生，实时RT-PCR结果显示，在SCAPs中，KDM2B的过表达抑制了软骨分化标志物COL1、COL2和SOX9的表达。相反，KDM2B的缺失增加了蛋白聚糖的产生，并抑制了COL1、COL2和SOX9的表达。总之，我们的结果表明KDM2B是SCAPs软骨分化的负调节因子，并表明抑制KDM2B可能会改善MSCs介导的软骨再生。
