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cDNA microarray analysis of gene expression changes induced by dexamethasone in cultured human trabecular meshwork
Takeshi Ishibashi1, Yasutaka Takagi, Kazuhiko Mori
1Departments of Ophthalmology, Kyoto Prefectural University of Medicine, Japan. tishibas@ophth.kpu-m.ac.jp
Investigative Ophthalmology & Visual Science
|November 28, 2002
Summary
Dexamethasone treatment significantly alters gene expression in human trabecular meshwork cells, upregulating genes like myocilin (MYOC). This provides insights into steroid-induced glaucoma development.
Area of Science:
- Ophthalmology
- Molecular Biology
- Genetics
Background:
- Steroid-induced glaucoma is a significant ocular condition.
- Understanding gene expression changes in the trabecular meshwork (TM) is crucial for elucidating disease mechanisms.
Purpose of the Study:
- To profile gene expression changes in cultured human TM cells induced by dexamethasone.
- To identify specific genes associated with the development of steroid-induced glaucoma.
Main Methods:
- Human TM cells from normal subjects were treated with dexamethasone or vehicle control.
- Gene expression profiling was performed using a human cDNA microarray containing 2400 genes.
- Upregulation and downregulation of gene expression were analyzed after 7 days of treatment.
Main Results:
- Dexamethasone treatment led to a twofold increase in 30 genes, with myocilin (MYOC), decorin, IGFBP2, ferritin L, and fibulin-1C being most significantly upregulated.
- Semiquantitative RT-PCR confirmed the upregulation of these key genes.
- A decrease in expression (downregulation) was observed in 34 genes, many with previously unreported changes.
Conclusions:
- cDNA microarray analysis is effective for studying gene expression in TM cells.
- The study confirms MYOC upregulation by dexamethasone and suggests other altered genes may contribute to reduced outflow facility.
- These findings offer new perspectives on the pathogenesis of steroid-induced glaucoma.