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A positive circuit of VEGF increases Glut-1 expression by increasing HIF-1α gene expression in human retinal endothelial cells

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Abstract

Treatment of human retinal microvascular endothelial cells (HRMECs) with vascular endothelial growth factor 165 (VEGF165) increased hypoxia-inducible factor 1α (HIF-1α), VEGF, and glucose transporter 1 (Glut-1) mRNA expression and Glut-1 protein localization to the membrane. In contrast, treatment of human retinal pigment epithelium cells with VEGF165 did not induce HIF-1α, VEGF, and Glut-1 gene expression. Microvascular endothelial cells are surrounded by astrocytic end feet in the retina. Astrocyte-derived A-kinase anchor protein 12 overexpression during hypoxia downregulated VEGF secretion, and this conditioned medium reduced VEGF and Glut-1 expression in HRMECs, suggesting that communications between astrocytes and endothelial cells may be the determinants of the blood vessel network. In HRMECs, HIF-1α small interfering RNA transfection blocked the VEGF165-mediated increase in VEGF and Glut-1 gene expression. Inhibition of protein kinase C (PKC) with inhibitor GF109203X or with a small interfering RNA targeting PKCζ attenuated the VEGF165-induced Glut-1 protein expression and VEGF and Glut-1 mRNA expression. In addition, results of an immunoprecipitation assay imply an interaction between VEGF receptor 2 (VEGFR2) and PKCζ in HRMECs. Therefore, VEGF secretion by hypoxic astrocytes may upregulate HIF-1α gene expression, inducing VEGF and Glut-1 expression via the VEGFR2–PKCζ axis in HRMECs.

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Acknowledgements

This paper was supported by Konkuk University in 2017. Dr. Irwin H. Gelman (Roswell Park Cancer Institute, Buffalo, NY, USA) kindly provided the full-length rat Akap12 cDNA, and Dr. Kyu-Won Kim (Seoul National University, Seoul, Republic of Korea) kindly gave us the subcloned vector (pcDNA3/AKAP12).

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  1. Department of Integrative Bioscience and Biotechnology, Konkuk University, Seoul, 05029, Republic of Korea

    Yoon Kyung Choi

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Choi, Y. A positive circuit of VEGF increases Glut-1 expression by increasing HIF-1α gene expression in human retinal endothelial cells. Arch. Pharm. Res. 40, 1433–1442 (2017). https://doi.org/10.1007/s12272-017-0971-5

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