Abstract
Purpose: :
Previously we showed the accelerated expression of angiogenetic genes (e.g. VEGF–A, VEGF–B, and αV integrin) in human RPE cell line (ARPE19) cultured on collagen I matrix (ARVO 2005, 3097). The purpose of this study is to demonstrate the pharmacologic regulation of angiogenetic gene expressions induced by collagen I.
Methods: :
Visually confluent ARPE19 cells were grown on collagen I coated flasks, and then serum was withdrawn for 48 hours. Alendronate, which is one of the bisphosphonate and known as an antagonist of αV integrin, was added to each dish (dose range 1µM to 1mM) and incubated for additional 24 hours. Total RNA was extracted and reverse transcribed. The gene expressions among each condition were compared using real–time PCR analysis.
Results: :
Alendronate inhibited the gene expression of αV integrin to 25% of control at 1mM. VEGF–A and VEGF–B was also down–regulated to 12% and 20% of control at a same dose, respectively.
Conclusions: :
Bisphosphonate inhibited the gene expression of αV integrin, VEGF–A and VEGF–B induced by collagen I. It suggests that the antagonists of αV integrin may interrupt the cascade of collagen I–induced angiogenetic gene expressions.
Keywords: retinal pigment epithelium • gene/expression • age-related macular degeneration