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VEGF-A regulates the expression of VEGF-C in human retinal pigment epithelial cells
  1. B Zhao1,*,
  2. A Ma1,
  3. J Cai1,
  4. M Boulton2
  1. 1School of Optometry and Vision Sciences, Cardiff University, Cardiff, UK
  2. 2Department of Ophthalmology and Visual Sciences, The University of Texas Medical Branch, Galveston, TX, USA
  1. Correspondence to: Professor Mike Boulton Department of Ophthalmology and Visual Sciences, The University of Texas Medical Branch, 301 University Blvd, Galveston, TX 77555-1106, USA; boultonm{at}utmb.edu

Abstract

Aim: To determine the expression and regulation of vascular endothelial growth factor C (VEGF-C), and its receptor VEGFR-3, in human retinal pigment epithelial (RPE) cells and to consider their angiogenic role in choroidal neovascularisation (CNV).

Method: The expression of VEGF-C and VEGFR-3 in cultured human RPE was confirmed by immunostaining, PCR, western blotting, and ELISA. Cultured RPE cells were exposed to VEGF-A and glucose and VEGF-C and VEGFR-3 changes in gene expression determined by RT-PCR. Secreted VEGF-C protein in conditioned media from RPE was examined by western blotting and ELISA analysis. The ability of VEGF-C to elicit tube formation in choroidal endothelial cells was assayed by an in vitro Matrigel model.

Result: VEGF-A and glucose upregulated VEGF-C mRNA expression and increased the secretion of VEGF-C protein into the culture medium. VEGF-A, but not glucose alone, stimulated VEGFR-3 mRNA expression. VEGF-C acted synergistically with VEGF-A to promote in vitro tube formation by choroidal endothelial cells.

Conclusion: VEGF-A has a critical role in the orchestration of VEGF-C expression in RPE cells and the synergistic action of VEGF-C with VEGF-A may play an important part in the aetiology of CNV.

  • AMD, age related macular degeneration
  • CNV, choroidal neovascularisation
  • ELISA, enzyme linked immunosorbent assay
  • FCS, fetal calf serum
  • GAPDH, glyceraldehyde-3-phosphate dehydrogenase
  • MAPKs, mitogen activated protein kinases
  • PCR, polymerase chain reaction
  • PDR, proliferative diabetic retinopathy
  • PKC, protein kinase C
  • RPE, retinal pigment epithelium
  • RT-PCR, reverse transcription polymerase chain reaction
  • VEGF, vascular endothelial growth factor
  • vascular endothelial growth factor
  • choroidal neovascularisation
  • vascular endothelial growth factor A
  • retinal pigment epithelial cells
  • choroidal endothelial cells
  • angiogenesis
  • AMD, age related macular degeneration
  • CNV, choroidal neovascularisation
  • ELISA, enzyme linked immunosorbent assay
  • FCS, fetal calf serum
  • GAPDH, glyceraldehyde-3-phosphate dehydrogenase
  • MAPKs, mitogen activated protein kinases
  • PCR, polymerase chain reaction
  • PDR, proliferative diabetic retinopathy
  • PKC, protein kinase C
  • RPE, retinal pigment epithelium
  • RT-PCR, reverse transcription polymerase chain reaction
  • VEGF, vascular endothelial growth factor
  • vascular endothelial growth factor
  • choroidal neovascularisation
  • vascular endothelial growth factor A
  • retinal pigment epithelial cells
  • choroidal endothelial cells
  • angiogenesis

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Footnotes

  • * Current address: Department of Biological Sciences, Lancaster University, Lancaster LA1 4YQ, UK.

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