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Vitreous Induces Components of the Prostaglandin E₂ Pathway in Human Retinal Pigment Epithelial Cells

From the Department of Pathology and Microbiology, University of South Carolina School of Medicine, Columbia, South Carolina.

PURPOSE. To investigate the alterations in gene expression when human retinal pigment epithelial (RPE) cells in culture are treated with vitreous as a model for the changes that occur in proliferative vitreoretinopathy.

METHODS. Human RPE cells were cultured with or without human vitreous or collagen. RNA was extracted and reverse transcribed. The RNAs expressed were compared by using DNA macroarrays. Messenger RNA levels were also measured using real-time reverse transcription polymerase chain reaction. Protein expression was examined by immunoblot analysis. Immunoassays were used to determine levels of prostaglandin E₂.

RESULTS. Vitreous treatment of RPE cells resulted in increased expression of two critical enzymes in the synthesis of prostaglandin E₂: membrane-associated prostaglandin E-synthase (mPGES) and cyclooxygenase (COX)-2. Increased levels of mPGES RNA and protein were still present at 48 hours of treatment, but the increase in COX-2 mRNA and protein was transient. The increase in the expression of mPGES was associated with an increase in the production of prostaglandin E₂ that was observed at 12 and 24 hours of treatment but not at 48 hours. Treatment with 100 µg collagen I per ml medium did not cause increased expression of mPGES and COX-2, even though both collagen- and vitreous-treatment caused a morphologic change in the RPE cells to a more fibroblast-like phenotype.

CONCLUSIONS. Treatment of human RPE cells with vitreous induces changes in gene expression that are indicative of an inflammatory response.

This article has been cited by other articles:

				R. Ganti, R. C. Hunt, S. K. Parapuram, and D. M. Hunt Vitreous Modulation of Gene Expression in Low-Passage Human Retinal Pigment Epithelial Cells Invest. Ophthalmol. Vis. Sci., April 1, 2007; 48(4): 1853 - 1863. [Abstract] [Full Text] [PDF]