Han Xiaofeng, Li Xinfeng, Zhong Guibin, Liu Zude
Department of Orthopaedics, Renji Hospital, School of Medicine, Shanghai Jiao Tong UniversityShanghai 200127, China.
Department of Orthopaedics, Renji Hospital South Campus, School of Medicine, Shanghai Jiao Tong UniversityShanghai 201112, China.
Am J Transl Res. 2017 Nov 15;9(11):4848-4855. eCollection 2017.
Bone marrow mesenchymal stem cells (BMSCs) are stem cells with multidirectional differentiation potential, which can be used as seed cells to repair and reconstruct many types of tissues and organs following injury or disease. Osteogenic differentiation involves a variety of pathway and factors, including cytokines, growth factors, and hormones. In the present study, we investigated the potential role of Dishevelled in osteogenic differentiation of BMSCs in induction medium containing the methyltransferase inhibitor 5-aza-2'-deoxycytidine. The expression of Dishevelled was analyzed using the reverse transcriptase-polymerase chain reaction (RT-PCR) and a Western blot. The methylation degree of the CpG island in the promoter region of the Dishevelled gene was analyzed, and protein expression levels of Wnt, Glycogen synthase kinase-3 (GSK3), axin, Dishevelled, and β-catenin were increased after the addition of the methyltransferase inhibitor. The expression of Dishevelled increased in accordance with the differentiation of osteoblasts, and the degree of methylation of the promoter affected its expression level. In conclusion, regulating the methylation degree of the Dishevelled gene promoter region appears to influence the expression of Dishevelled and therefore the osteogenic differentiation of BMSCs.
骨髓间充质干细胞(BMSCs)是具有多向分化潜能的干细胞,可作为种子细胞用于修复和重建损伤或疾病后的多种组织和器官。成骨分化涉及多种途径和因子,包括细胞因子、生长因子和激素。在本研究中,我们在含有甲基转移酶抑制剂5-氮杂-2'-脱氧胞苷的诱导培养基中,研究了Dishevelled在BMSCs成骨分化中的潜在作用。使用逆转录聚合酶链反应(RT-PCR)和蛋白质印迹法分析Dishevelled的表达。分析了Dishevelled基因启动子区域CpG岛的甲基化程度,添加甲基转移酶抑制剂后,Wnt、糖原合酶激酶-3(GSK3)、axin、Dishevelled和β-连环蛋白的蛋白质表达水平升高。Dishevelled的表达随着成骨细胞的分化而增加,启动子的甲基化程度影响其表达水平。总之,调节Dishevelled基因启动子区域的甲基化程度似乎会影响Dishevelled的表达,从而影响BMSCs的成骨分化。