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VEGF-A regulates the expression of VEGF-C in human retinal pigment epithelial cells

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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