Regulation of human corneal epithelial cell proliferation and apoptosis by dexamethasone

Invest Ophthalmol Vis Sci. 2000 Dec;41(13):4133-41.

Abstract

Purpose: To investigate whether human corneal epithelial cells express the glucocorticoid receptor (GR) and to assess the influence of dexamethasone (DEX) on these cells.

Methods: Human corneal epithelial cells were cultured in medium supplemented with various concentrations of DEX (ranging from 10(-10) to 10(-4) M). Cell proliferation was analyzed by 3-(4, 5-dimethylthiazol-2-yl)-5-(3-carboxy-methoxyphenyl)-2-(4-sulfop henyl) -2H-tetrazolium inner salt (MTS) assay at 2, 4, and 6 days of culture. Apoptosis was studied by nucleus labeling using a fluorescent dye and immunostaining by APO 2.7 at 6 days of culture. GR mRNA was detected in corneal epithelium and cultured corneal epithelial cells by means of reverse transcription-polymerase chain reaction (RT-PCR). Immunocytochemical staining of the epithelial cells was performed with a monoclonal anti-human GR.

Results: RT-PCR and immunocytochemistry showed the expression of GR (mRNA and protein) in corneal epithelial cells. DEX significantly increased corneal epithelial cell proliferation with concentrations ranging from 10(-10) to 10(-6) M, with a maximum effect at 10(-7) M (P < 0.005). However, DEX also induced apoptosis of cultured corneal epithelial cells at any concentration used.

Conclusions: These results indicate that human corneal epithelial cells express the GR and proliferate in response to DEX stimulation which also induces corneal epithelial cell apoptosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis / drug effects*
  • Cell Division / drug effects*
  • Cells, Cultured
  • DNA Primers / chemistry
  • Dexamethasone / pharmacology*
  • Epithelium, Corneal / cytology*
  • Epithelium, Corneal / drug effects
  • Epithelium, Corneal / metabolism*
  • Gene Expression
  • Glucocorticoids / pharmacology*
  • Humans
  • Immunoenzyme Techniques
  • Methylphenazonium Methosulfate / metabolism
  • RNA, Messenger / metabolism
  • Receptors, Glucocorticoid / biosynthesis
  • Receptors, Glucocorticoid / genetics*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Tetrazolium Salts / metabolism
  • Thiazoles / metabolism

Substances

  • DNA Primers
  • Glucocorticoids
  • RNA, Messenger
  • Receptors, Glucocorticoid
  • Tetrazolium Salts
  • Thiazoles
  • 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium
  • Methylphenazonium Methosulfate
  • Dexamethasone