May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
Small GTPase Rho Regulates Corneal Epithelial Wound Healing Independently of EGFR
Author Affiliations & Notes
  • J. Yin
    Anatomy & Cell Biology, Wayne State Univ Sch of Med, Detroit, Michigan
  • F.-S. X. Yu
    Anatomy & Cell Biology, Wayne State Univ Sch of Med, Detroit, Michigan
  • Footnotes
    Commercial Relationships  J. Yin, None; F.X. Yu, None.
  • Footnotes
    Support  NIH/NEI EY10869 and 14080, MidWest Eye banks Student Stipend
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 3378. doi:https://doi.org/
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    • Get Citation

      J. Yin, F.-S. X. Yu; Small GTPase Rho Regulates Corneal Epithelial Wound Healing Independently of EGFR. Invest. Ophthalmol. Vis. Sci. 2008;49(13):3378. doi: https://doi.org/.

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

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Abstract

Purpose: : To determine the role of small GTPase Rho and its relation with epidermal growth factor receptor (EGFR) in mediating corneal epithelial wound healing.

Methods: : Rho activity in THCE cells, an SV40-immortalized human corneal epithelial cell (HCEC) line, was assessed by Western blotting. Rho functions were inhibited with specific inhibitor exoenzyme C3 (C3) and confirmed by knockdown with small interference RNA (siRNA) transfection. Effects of Rho inhibition on wound healing were determined in porcine corneal organ culture and HCEC scratch wound models. Effects of C3 on cell proliferation and focal adhesion formation were determined by BrdU incorporation assay and immunocytochemistry, respectively.

Results: : Wounding, lysophosphatidic acid and heparin-binding EGF-like growth factor (HB-EGF) induced rapid and strong RhoA activation. HB-EGF-, but not wounding-, enhanced RhoA activity was sensitive to EGFR inhibition. In corneal organ and cell culture models, C3 attenuated both spontaneous and HB-EGF-induced wound closures, confirmed by delayed wound healing in cells transfected with RhoA siRNA. C3 also retarded spontaneous wound healing in the presence of hydroxyurea, a cell cycle blocker. C3 significantly reduced the number of BrdU-positive cells near the leading edge. Treatment with C3 resulted in the disruption of the cortical actin cytoskeleton and in the disappearance of paxillin-containing focal adhesion and lamellipodia.

Conclusions: : Wounding induces RhoA activation via an EGFR-independent pathway. Rho activity is required for modulating both cell migration and proliferation, through cytoskeleton reorganization and focal adhesion formation in response to wounding.

Keywords: cornea: epithelium • wound healing • signal transduction 
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