[Frontiers in Bioscience S5, 661-671, January 1, 2013]

Migration of retinal pigment epithelial cells is EGFR/PI3K/AKT dependent

Lei Zhang1, Fenghua Wang2, Yuan Jiang1,2, Shan Xu1,2,Fengqing Lu1,2,Wenqiu Wang1,2, Xiangjun Sun3, Xiaodong Sun1,2

1Department of Ophthalmology, Shanghai First People's Hospital Affiliated Shanghai Jiao Tong University, 100 Haining road, Shanghai, PR China, 200080, 2Shanghai Eye Research Institute, 100 Haining road, Shanghai, PR China, 200080, 3School of Agriculture and Biology, Shanghai Jiao Tong University, 800 Dongchuan RD. Minhang District, Shanghai, PR China, 200080


1. Abstract
2. Introduction
3. Materials and methods
3.1. Reagents
3.2. Cell culture
3.3. MTS cell proliferation assay
3.4. In vitro wound healing assay
3.5. Transwell cell migration Assay
3.6. Western blot
3.7. Immunofluorescence
3.8. Statistical analysis
4. Results
4.1. ARPE-19 cell proliferation assay
4.2. EGF Enhances the migration of ARPE-19 cells via EGFR
4.3. PI3-Kinase activation is required for EGF-mediated ARPE-19 cell migration
4.4. AKT is involved in EGF-mediated ARPE-19 cell migration
4.5. Comparison of the roles of EGFR/PI3K/AKT on ARPE-19 cell migration
5. Discussion
6. Acknowledgements
7. References


Abnormal migration of retinal pigment epithelium (RPE) contributes to a variety of disorders such as proliferative vitreoretinopathy. Here, the effect of epidermal growth factor (EGF), and signaling by its receptor (ERGR)/phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) on RPE cell migration was studied. The in vitro wound healing and migration of the human RPE cell line, ARPE19 cell was accelerated, in a dose dependent manner, in response to EGF stimulation, while pretreatment with EGFR, PI3K or AKT inhibitor, inhibited both events. Exposure of cells to EGF activated the AKT phosphorylation, whereas EGFR and PI3K inhibitors blocked EGF-induced AKT phosphorylation in a dose-dependent manner. These data suggest that EGF mediate ARPE-19 cell migration through EGFR/PI3K/AKT signaling pathway.