1. GEP100/ARF6 regulates VEGFR2 signaling to facilitate high-glucose-induced epithelial-mesenchymal transition and cell permeability in retinal pigment epithelial cells
- Author
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Shan-Shan Chen, Shu-Hua Fu, Zhi-Peng You, Fan Xiong, Ting Liu, Shu-Rong Li, and Zhong-Yi Yang
- Subjects
0301 basic medicine ,Cell Membrane Permeability ,Epithelial-Mesenchymal Transition ,Physiology ,Cell ,Retinal Pigment Epithelium ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,Pigment ,0302 clinical medicine ,medicine ,Animals ,Guanine Nucleotide Exchange Factors ,Humans ,Epithelial–mesenchymal transition ,Cell permeability ,Cells, Cultured ,Retinal pigment epithelium ,Transition (genetics) ,ADP-Ribosylation Factors ,Chemistry ,Retinal ,Cell Biology ,Diabetic retinopathy ,medicine.disease ,Vascular Endothelial Growth Factor Receptor-2 ,Cell biology ,Mice, Inbred C57BL ,Glucose ,030104 developmental biology ,medicine.anatomical_structure ,ADP-Ribosylation Factor 6 ,030220 oncology & carcinogenesis ,visual_art ,visual_art.visual_art_medium ,Signal Transduction - Abstract
Cell permeability and epithelial-mesenchymal transition (EMT) were found to be enhanced in diabetic retinopathy, and the aim of this study was to investigate the underlying mechanism. ARPE-19 cell line or primary retinal pigment epithelial (RPE) cells were cultured under high or normal glucose conditions. Specific shRNAs were employed to knock down ADP-ribosylation factor 6 (ARF6), GEP100, or VEGF receptor 2 (VEGFR2) in ARPE-19 or primary RPE cells. Cell migration ability was measured using Transwell assay. Western blotting was used to measure indicated protein levels. RPE cells treated with high glucose showed increased cell migration, paracellular permeability, EMT, and expression of VEGF. Knockdown of VEGFR2 inhibited the high-glucose-induced effects on RPE cells via inactivation of ARF6 and MAPK pathways. Knockdown ARF6 or GEP100 led to inhibition of high-glucose-induced effects via inactivation of VEGFR2 pathway. Knockdown of ARF6, but not GEP100, decreased high-glucose-induced internalization of VEGFR2. High-glucose enhances EMT and cell permeability of RPE cells through activation of VEGFR2 and ARF6/GEP100 pathways, which form a positive feedback loop to maximize the activation of VEGF/VEGFR2 signaling.
- Published
- 2019