May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
Oxidative Stress Induces Epithelial-Mesenchymal Transition-Like Changes in Cultured Corneal Epithelial Cells
Author Affiliations & Notes
  • N. Kato
    Keio Univ School of Medicine, Shinjuku-ku, Japan
    Ophthalmology,
  • S. Shimmura
    Keio Univ School of Medicine, Shinjuku-ku, Japan
    Ophthalmology,
  • T. Kawakita
    Keio Univ School of Medicine, Shinjuku-ku, Japan
    Ophthalmology,
  • S. Yoshida
    Keio Univ School of Medicine, Shinjuku-ku, Japan
    Ophthalmology,
  • H. Okano
    Keio Univ School of Medicine, Shinjuku-ku, Japan
    Physiology,
  • K. Tsubota
    Keio Univ School of Medicine, Shinjuku-ku, Japan
    Ophthalmology,
  • Footnotes
    Commercial Relationships  N. Kato, None; S. Shimmura, None; T. Kawakita, None; S. Yoshida, None; H. Okano, None; K. Tsubota, None.
  • Footnotes
    Support  Advanced and Innovational Research Program in Life Sciences from the Ministry of Education, Culture, Sports, Science and Technology
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 5038. doi:https://doi.org/
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      N. Kato, S. Shimmura, T. Kawakita, S. Yoshida, H. Okano, K. Tsubota; Oxidative Stress Induces Epithelial-Mesenchymal Transition-Like Changes in Cultured Corneal Epithelial Cells. Invest. Ophthalmol. Vis. Sci. 2008;49(13):5038. doi: https://doi.org/.

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

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Abstract

Purpose: : Pteryium is known to correlate with cumurative ultraviolet exposure on the corneal limbus. We previously reported epithelial cells at the leading edge of pterygial head lost epithelial characteristics, such as E-cadherin expression, and simultaneously upregulated mesenchymal markers, such as α-smooth muscle actin (SMA) and vimentin (Kato et al, IOVS 2007). This phenomenon is called epithelial-mesenchymal transition (EMT). We speculated that oxidative stress caused by ultraviolet exposure may induce EMT in the pterygial epithelium. Therefore, we investigated whether oxidative stress induces epithelial-mesenchymal transition (EMT) in cultured corneal epithelial cells.

Methods: : TKE2 cells (mouse corneal epithelial cell line) were harvested in 75 mm2 flasks and cultured in 5% CO2 at 37oC using the defined KSFM medium with or without hydrogen peroxide (H2O2; 10~50 µM). After 7 days, RT-PCR, western blotting for α-SMA, E-cadherin, and immunohistochemistry for α-SMA, E-cadherin, and β-catenin were performed.

Results: : By phasecontrast microscopy, TKE2 cells revealed enlarged, flattened and fusiform-like elongated contour after 7 days H2O2 stimulation. RT-PCR and western blotting showed upregulated expression of α-SMA in the cells stimulated with H2O2. Immunohistochemistry showed declined membrane staining for E-cadherin and β-catenin and increased staining for α-SMA in the H2O2 stimulated cells.

Conclusions: : H2O2 caused EMT-like changes in cultured corneal epithelial cells. The present results may indicate that oxidative stress by ultraviolet exposure may cause EMT in the pathogenesis of pterygium.

Keywords: EMT (epithelial mesenchymal transition) • oxidation/oxidative or free radical damage • cornea: epithelium 
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