Takehara Toshiyuki, Teramura Takeshi, Onodera Yuta, Frampton John, Fukuda Kanji
Division of Cell Biology for Regenerative Medicine, Institute of Advanced Clinical Medicine, Kindai University Faculty of Medicine, 377-2 Ohnohigashi, Osaka-sayama, Osaka, Japan 5898511.
School of Biomedical Engineering, Dalhousie University, Halifax, Nova Scotia, Canada B3H 4R2 1-902-494-4175.
Sci Rep. 2015 Sep 30;5:14722. doi: 10.1038/srep14722.
The cell adhesion molecule Cadherin 2 (Cdh2) plays important roles in somatic cell adhesion, proliferation and migration. Cdh2 is also highly expressed in mouse epiblast stem cells (mEpiSCs), but its function in these cells is unknown. To understand the function of Cdh2 in mEpiSCs, we compared the expression of pluripotency-related genes in mEpiSCs and mouse embryonic stem cells (mESCs) after either Cdh2 knockdown or Cdh2 over-expression. Introduction of specific siRNA against Cdh2 led to attenuation of pluripotency-related genes. Pluripotent gene expression was not recovered by over-expression of Cdh1 following Cdh2 knockdown. Western blot analysis and co-immunoprecipitation assays revealed that Cdh2 stabilizes FGFR1 in mEpiSCs. Furthermore, stable transfection of mESCs with Cdh2 cDNA followed by FGF2 supplementation accelerated cell differentiation. Thus, Cdh2 contributes to the establishment and maintenance of FGF signaling-dependent self-renewal in mEpiSCs through stabilization of FGFR1.
细胞粘附分子钙粘蛋白2（Cdh2）在体细胞粘附、增殖和迁移中发挥重要作用。Cdh2在小鼠上胚层干细胞（mEpiSCs）中也高度表达，但其在这些细胞中的功能尚不清楚。为了了解Cdh2在mEpiSCs中的功能，我们比较了Cdh2敲低或过表达后mEpiSCs和小鼠胚胎干细胞（mESCs）中多能性相关基因的表达。引入针对Cdh2的特异性siRNA导致多能性相关基因的表达减弱。Cdh2敲低后，Cdh1的过表达未能恢复多能基因的表达。蛋白质免疫印迹分析和免疫共沉淀试验表明，Cdh2在mEpiSCs中稳定FGFR1。此外，用Cdh2 cDNA稳定转染mESCs，然后补充FGF2可加速细胞分化。因此，Cdh2通过稳定FGFR1有助于在mEpiSCs中建立和维持FGF信号依赖的自我更新。
