摘要
背景:间充质干细胞移植后的归巢能力关系到细胞移植的疗效,研究其趋化和迁移的调控将有助于提高间充质干细胞临床应用的价值。目的:观察Cdc42在人脐带间充质干细胞定向迁移中的作用。方法:首先以组织块法分离培养人脐带间充质干细胞,与炎性因子肿瘤坏死因子α、白细胞介素1β、转化生长因子β共培养后Western检测Cdc42表达变化。化学合成Cdc42的干扰RNA,转染细胞后分别采用Transwell和Matrigel胶观察细胞迁移和黏附能力。应用Western检测Cdc42下游靶分子ERK1/2的变化。结果与结论:与炎性因子共培养后的人脐带间充质干细胞中Cdc42表达明显增加,接近无因子对照组的2倍水平。siRNA下调Cdc42的表达能够显著抑制人脐带间充质干细胞的迁移和黏附,且其下游信号分子ERK1/2的表达以及磷酸化水平均相应受到抑制。提示体外培养环境下Cdc42参与了人脐带间充质干细胞的趋化过程。
BACKGROUND：The homing ability of mesenchymal stem cells is closely associated with the effects of celltransplantation. Clarifying the mechanism of chemotaxis and migration wil contribute to enhance the clinical application of mesenchymal stem cells. OBJECTIVE：To investigate the effect of Cdc42 in the homing of human umbilical cord mesenchymal stem cells. METHODS：First, mesenchymal stem cells were isolated from human umbilical cord, and co-cultured with tumor necrosis factorα, interleukin-1β, and transforming growth factorβ. Western blot assay was used to test the level of Cdc42. Besides, Cdc42 siRNA was synthesized by chemical method to transfect the cells, and cellmigration and adhesion were measured by Transwel and Matrigel separately. Meanwhile, the activity of signal molecule, extracellular regulated protein kinase 1/2, was evaluated by western blot. RESULTS AND CONCLUSION：The results indicated that the inflammation factors induced the highly expression of Cdc42 in human umbilical cord mesenchymal stem cells, almost double level to controls. siRNA notably inhibited the migration and adhesion of human umbilical cord mesenchymal stem cells through Cdc42 down-regulation, and the extracellular regulated protein kinase 1/2 and phosphorylation form were also decreased simultaneously. In a word, we speculate Cdc42 plays a role in the chemotaxis of human umbilical cord mesenchymal stem cells in vitro.
出处
《中国组织工程研究》
CAS
CSCD
2014年第23期3616-3621,共6页
Chinese Journal of Tissue Engineering Research
基金
国家自然科学基金青年项目(81001106)~~
