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Amanda-Jayne F. Carr, Anthony A. Vugler, Stephen E. Moss, John Greenwood, Pete Coffey; Changes in Extracellular Matrix Components of ARPE-19 Cells by Fenretinide. Invest. Ophthalmol. Vis. Sci. 2011;52(14):870.
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Components of the extracellular matrix have an important role in many developmental and regenerative processes, and have also been implicated in neural retinal regeneration from retinal pigment epithelium. We have previously shown that the human retinal pigment epithelial cell line ARPE-19 is capable of differentiating towards a neuroretinal-like phenotype in culture after treatment with fenretinide. Here we have examined the effect of fenretinide on extracellular matrix components in ARPE-19 cells.
ARPE-19 cells were cultured in DMEM/F12 (1:1) supplemented with 3% charcoal/dextran treated fetal calf serum, 1x penicillin/streptomycin and either 3uM fenretinide or an equal volume of DMSO as a control for 7 days. Cells were then processed for analysis by microarray hybridisation, Q-PCR, immunocytochemistry and Western blot.
Our experiments have identified a number of extracellular matrix proteins, such as fibronectin, laminin, collagen type IV and heparin sulphate proteoglycan, whose expression is altered after treatment with fenretinide in ARPE-19 cells. Furthermore, these changes were accompanied by an increase in tissue plasminogen activator (PLAT), urokinase plaminogen activator (PLAU) and its receptor (PLAUR).
Changes in the expression levels of extracellular matrix components and the proteolytic pathway associated with its degradation suggests that reorganisation of the ECM may be an important factor in the differentiation of human RPE cells towards a neuroretinal-like phenotype.
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