人E-钙粘素融合蛋白基质激活脐带间充质干细胞EGF受体及其功能的研究  

Human E-cadherin Fusion Protein Matrix Activates EGF Receptor and its Function in Human Umbilical Cord Mesenchymal Stem Cells

作　　者：曹磊 张妍[1] 杨军[1] Cao Lei;Zhang Yan;Yang Jun(College of Life Sciences,Nankai University,Tianjin 300071,China)

机构地区：[1]南开大学生命科学学院,天津300071

出　　处：《南开大学学报（自然科学版）》2020年第2期91-99,共9页Acta Scientiarum Naturalium Universitatis Nankaiensis

基　　金：国家自然科学基金(31771067);国家自然科学基金青年项目(81703094);国家重点研发计划(2016YFC1101304);天津市应用基础与前沿技术研究计划重点资助项目(17JCZDJC37600)。

摘　　要：上皮细胞钙粘素(E-cadherin)介导细胞间粘附连接,通过多级信号转导调控细胞粘附、增殖、迁移等多种细胞行为.本实验室前期研究中成功构建了基于E-cadherin胞外域的E-cad-Fc融合蛋白作为人间充质干细胞(hMSCs)的培养基质,其能够激活上皮细胞生长因子受体(Epidermal growth factor receptor,EGFR)信号通路,促进hMSCs增殖,但目前为止尚不明确E-cad-Fc基质能否替代EGF在hMSCs培养过程中发挥特异性功能.因此,本研究首先利用基因芯片技术分析了E-cad-Fc对EGF相关信号通路的影响,并筛选出显著差异表达的基因.利用western blot评价了E-cad-Fc和EGF在短时间内对hMSCs中EGFR的激活,并初步评价了两者对hMSCs定向诱导分化的影响.研究结果表明,E-cad-Fc基质可替代EGF持续性激活EGFR的磷酸化,并在定向诱导分化过程中促进hMSCs的转胚层进程.该研究为探讨钙粘素基质对hMSCs细胞行为的调控作用及其信号转导机制奠定了基础.E-cadherin mediates the cell adhesion junction,and regulates the cell adhesion, proliferation,migration and other cellular behaviors through multilevel signal transduction. In our previous studies, Ecad-Fc fusion protein that based on the extracellular domain of E-cadherin was successfully constructed as the culture matrix of human mesenchymal stem cells(hMSCs). It could activate the epidermal growth factor receptor(EGFR) signaling pathway, promote the proliferation of hMSCs. However,it was still unclear whether E-cad-Fc matrix can replace EGF and played a specific role in hMSCs culture. Therefore, this study first applied DNA chip to analyze the influence of E-cad-Fc on EGF-related signaling pathways,and screened out the genes with significantly different expressions. Western blotting was used to evaluate the activation of EGFR in hMSCs by E-cad-Fc and EGF in a short period of time, and the effects of E-cad-Fc and EGF on the directed differentiation of h MSCs were preliminarily evaluated. The results showed that E-cad-Fc could replace EGF to continuously activate EGFR phosphorylation and promote the transdermal process of h MSCs in the process of directed differentiation. This study laid a foundation for exploring the regulation of cadherin matrix on the behavior of hMSCs and its signal transduction mechanism.

关 键 词：E-cad-Fc融合蛋白 EGFR信号通路 间充质干细胞 基因芯片 

分 类 号：R318[医药卫生—生物医学工程]