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Vitreous induces components of the prostaglandin E2 pathway in human retinal pigment epithelial cells.
- Source :
-
Investigative ophthalmology & visual science [Invest Ophthalmol Vis Sci] 2003 Apr; Vol. 44 (4), pp. 1767-74. - Publication Year :
- 2003
-
Abstract
- 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.<br />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 E(2).<br />Results: Vitreous treatment of RPE cells resulted in increased expression of two critical enzymes in the synthesis of prostaglandin E(2): 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 E(2) that was observed at 12 and 24 hours of treatment but not at 48 hours. Treatment with 100 microg 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.<br />Conclusions: Treatment of human RPE cells with vitreous induces changes in gene expression that are indicative of an inflammatory response.
- Subjects :
- Cells, Cultured
Collagen Type I physiology
Cyclooxygenase 2
Humans
Intramolecular Oxidoreductases metabolism
Isoenzymes metabolism
Membrane Proteins
Oligonucleotide Array Sequence Analysis
Pigment Epithelium of Eye cytology
Prostaglandin-E Synthases
Prostaglandin-Endoperoxide Synthases metabolism
RNA, Messenger metabolism
Reverse Transcriptase Polymerase Chain Reaction
Dinoprostone biosynthesis
Gene Expression physiology
Intramolecular Oxidoreductases genetics
Isoenzymes genetics
Pigment Epithelium of Eye metabolism
Prostaglandin-Endoperoxide Synthases genetics
Vitreous Body physiology
Subjects
Details
- Language :
- English
- ISSN :
- 0146-0404
- Volume :
- 44
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- Investigative ophthalmology & visual science
- Publication Type :
- Academic Journal
- Accession number :
- 12657620
- Full Text :
- https://doi.org/10.1167/iovs.02-0528