May 2004
Volume 45, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2004
Glucocorticoid Induced Changes in Gene Expression in Human Lens Epithelial Cells.
Author Affiliations & Notes
  • V.K. Gupta
    Biochemistry & Molecular Biology, UMDNJ– Graduate School of Biomedical Sciences, Newark, NJ
  • A.T. Galante
    Center for Applied Genomics, PHRI, Newark, NJ
  • P. Soteropoulos
    Center for Applied Genomics, PHRI, Newark, NJ
    Microbiology & Molecular Genetics,
    UMDNJ–New Jersey Medical School, Newark, NJ
  • B.J. Wagner
    Biochemistry & Molecular Biology, UMDNJ– Graduate School of Biomedical Sciences, Newark, NJ
    Ophthamology,
    UMDNJ–New Jersey Medical School, Newark, NJ
  • Footnotes
    Commercial Relationships  V.K. Gupta, None; A.T. Galante, None; P. Soteropoulos, None; B.J. Wagner, None.
  • Footnotes
    Support  NIH Grant EY02299
Investigative Ophthalmology & Visual Science May 2004, Vol.45, 2650. doi:
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      V.K. Gupta, A.T. Galante, P. Soteropoulos, B.J. Wagner; Glucocorticoid Induced Changes in Gene Expression in Human Lens Epithelial Cells. . Invest. Ophthalmol. Vis. Sci. 2004;45(13):2650.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Abstract: : Purpose: Prolonged use of glucocorticoids can lead to the formation of a cataract, however the mechanism is not known. We recently reported the presence of the functional glucocorticoid receptor in immortalized cultured mammalian lens epithelial cells (LECs), but the biological effect is not known. This study seeks to examine glucocorticoid induced changes in gene expression in LECs and to determine if freshly isolated human LECs respond to glucocorticoid treatment. Methods: RNA isolated from HLE B–3 cells treated with dexamethasone (DEX) or vehicle was used to analyze global changes in gene expression by microarray hybridization. The experiment was done in triplicate. Data and cluster analyses were performed using Microarray Suite 5.0 and Genespring 6.1. Capsulorhexis specimens obtained in surgery from eyes with cataract were cultured. Primary lens and HLE B–3 cultures were transfected with pGRE.Luc, which drives the expression of firefly luciferase, and treated with DEX or vehicle. Results: Microarray data revealed that 397 transcripts are upregulated and 4 transcripts are downregulated by 2 fold or more in LECs treated with DEX compared to vehicle treated cells. The majority of upregulated transcripts were found to be involved in cell growth and maintenance, cell communication, and signal transduction. Transfected primary human LEC cultures treated with DEX demonstrated a glucocorticoid response with a 2.5 to 15 fold increase in firefly luciferase activity over controls. HLE B–3 cells showed a 2 to 5 fold increase in luciferase activity over controls. Conclusions: Microarray studies demonstrate that glucocorticoids modulate gene expression in immortalized human LECs and show novel changes that may lead to a better understanding of pathways involved in a lens glucocorticoid response. The activation of a GRE reporter gene in primary human LEC cultures demonstrates that the glucocorticoid receptor is functional in non–immortalized human lens cells. This study demonstrates that primary lens cultures and microarray technology can be used to determine pathways involved in the formation of a steroid induced cataract.

Keywords: corticosteroids • gene microarray • gene/expression 
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