Eric R James1, Victor M Fresco, Lorie L Robertson. 1. Department of Ophthalmology, Storm Eye Institute, Medical University of South Carolina, Charleston, SC 29403, USA. jameser@musc.edu
Abstract
AIMS: Clinically, steroid use is accompanied by a risk for posterior subcapsular cataracts (PSCs). PSC possibly involves perturbation of lens epithelial cell proliferation and differentiation; however, the underlying mechanism is unknown. In this study, we aimed to characterize changes in gene expression in human lens epithelial cells exposed to glucocorticoid using DNA microarray hybridization. METHODS: Human lens epithelial cells (HLE B-3) were treated with dexamethasone (1 microM) for 24 or 48 hours or with vehicle (0.01% dimethylsulfoxide) and the derived cRNA was hybridized to U133A microarrays. Data were processed using the Affymetrix program Micro Array Suite, and ontological analyses were performed using the software dChip, filtering to exclude transcripts up- or down-regulated by less than 2-fold. RESULTS: At 24 hours, 57 transcripts were upregulated relative to controls by greater than 2- fold and 50 were downregulated by greater than 2-fold. At 48 hours, 92 transcripts were upregulated and 42 were downregulated. Twenty-two upregulated and 2 downregulated transcripts were shared between the 24-hour and 48-hour data sets. The predominant ontological groupings were: signal transduction, transcription factor activity, cytoskeleton/ECM/adhesion, transport, and cell cycle/development. Alternate ontological interpretations are possible. CONCLUSIONS: The data indicate steroid treatment of lens epithelial cells is associated with significant changes in gene expression in several functional categories and these include transcripts related to cell proliferation.
AIMS: Clinically, steroid use is accompanied by a risk for posterior subcapsular cataracts (PSCs). PSC possibly involves perturbation of lens epithelial cell proliferation and differentiation; however, the underlying mechanism is unknown. In this study, we aimed to characterize changes in gene expression in human lens epithelial cells exposed to glucocorticoid using DNA microarray hybridization. METHODS:Human lens epithelial cells (HLE B-3) were treated with dexamethasone (1 microM) for 24 or 48 hours or with vehicle (0.01% dimethylsulfoxide) and the derived cRNA was hybridized to U133A microarrays. Data were processed using the Affymetrix program Micro Array Suite, and ontological analyses were performed using the software dChip, filtering to exclude transcripts up- or down-regulated by less than 2-fold. RESULTS: At 24 hours, 57 transcripts were upregulated relative to controls by greater than 2- fold and 50 were downregulated by greater than 2-fold. At 48 hours, 92 transcripts were upregulated and 42 were downregulated. Twenty-two upregulated and 2 downregulated transcripts were shared between the 24-hour and 48-hour data sets. The predominant ontological groupings were: signal transduction, transcription factor activity, cytoskeleton/ECM/adhesion, transport, and cell cycle/development. Alternate ontological interpretations are possible. CONCLUSIONS: The data indicate steroid treatment of lens epithelial cells is associated with significant changes in gene expression in several functional categories and these include transcripts related to cell proliferation.
Authors: Kimberly F Whelan; Kayla Stratton; Toana Kawashima; John W Waterbor; Robert P Castleberry; Marilyn Stovall; Charles A Sklar; Roger J Packer; Pauline Mitby; Candice L Aitken; Julie Blatt; Leslie L Robison; Ann C Mertens Journal: Pediatr Blood Cancer Date: 2010-01 Impact factor: 3.167