Abstract

PURPOSE
Inhibition of VEGF in the eye is an important treatment modality for reducing proliferation and migration of retinal pigment epithelium (RPE) in age-related macular degeneration (AMD). Additionally, previous studies suggest calcium-independent phospholipase A(2) group VIA (iPLA(2)-VIA) to be a potential regulator of cell proliferation and migration, and evidence show abundant expression of iPLA(2)-VIA in RPE cells. The aim of the present study was to evaluate the potential role of iPLA(2)-VIA in VEGF-induced proliferation and migration of RPE cells.
MATERIALS AND METHODS
The human RPE cell line, ARPE-19, was used in all assays. To explore the role of iPLA(2)-VIA in VEGF-induced RPE proliferation and migration, iPLA(2)-VIA inhibition by the iPLA(2)-VIA specific inhibitor, bromoenol lactone, was done. RPE cell proliferation and migration were evaluated by measurements of incorporated radioactive thymidine in DNA and by a Boyden chamber technique, respectively. A luciferase assay monitored the VEGF-induced iPLA(2)-VIA transcriptional activity. Western blot analysis and an activity assay were used to detect the protein levels and activity of iPLA(2)-VIA respectively after treatment with VEGF.
RESULTS
RPE cells treated with VEGF showed significant increased proliferation and migration. Furthermore, inhibition of iPLA(2)-VIA significantly reduced the spontaneous proliferation and migration as well as the VEGF-induced proliferation and migration. Finally, inhibition of iPLA(2)-VIA reduced the VEGF-induced iPLA(2)-VIA-activity, -protein level, and -promoter activity.
CONCLUSIONS
A significant interaction between VEGF and iPLA(2)-VIA in the regulation of RPE cells appears to be relevant in elucidating the exact mechanisms of action in the proliferative and migratory phenotype of RPE cells in AMD.

Links

Publisher Full Text

Authors

Kehler AK, Andersen C, Andreasen JR, Vohra R, Junker N, Poulsen KA, Kolko M

Institution

Department of Neuroscience and Pharmacology, The Panum Institute, University of Copenhagen, Copenhagen, Denmark. akkehler@yahoo.com

Source

Current eye research 37:6 2012 Jun pg 500-7

MeSH

Blotting, Western
Cell Line
Cell Movement
Cell Proliferation
DNA Replication
Group VI Phospholipases A2
Humans
Naphthalenes
Phosphodiesterase Inhibitors
Pyrones
Retinal Pigment Epithelium
Transcriptional Activation
Up-Regulation
Vascular Endothelial Growth Factor A

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

22577768