title: Effect of EGF-induced HDAC6 activation on corneal epithelial wound healing.
abstract: OBJECTIVE
  Epidermal growth factor (EGF) stimulates migration in corneal epithelial wound healing. The purpose of this study was to investigate the effect of EGF-induced alpha-tubulin deacetylation through activating HDAC6 on migration in corneal epithelial wound healing.
  
  
  METHODS
  Human corneal epithelial (HCE) cells were cultured in DMEM/F12 medium containing 10% FBS in a 37 degrees C incubator supplied with 5% CO(2). Western blot analysis was used to determine protein expression. Activity of HDAC6 was suppressed by trichostatin A (TSA) and by siRNA specific to HDAC6. Corneal epithelial cell migration was measured by using scratch-induced directional migration assay in cultured cells and by corneal epithelial debridement using a mouse whole-eye organ culture model.
  
  
  RESULTS
  The authors found EGF stimulated corneal epithelial cell migration in wound healing by enhancing HDAC6 activity, resulting in the deacetylation of alpha-tubulin. EGF stimulated HDAC6 enzymatic activity and protein translocation toward the leading edge of the cell. Inhibition of HDAC6 activity by TSA significantly suppressed EGF-induced cell migration and delayed EGF-induced wound healing in epithelially debrided mouse corneas. In the meantime, knockdown of HDAC6 mRNA with specific siRNA effectively abolished EGF-induced deacetylation of alpha-tubulin, resulting in the inhibition of cell migration.
  
  
  CONCLUSIONS
  These results reveal an important mechanism that involves EGF-induced HDAC6 activation and alpha-tubulin deacetylation, subsequently affecting corneal epithelial migration in the wound-healing process.
authors.affiliation: Division of Molecular Medicine, Department of Medicine, David Geffen School of Medicine, University of California Los Angeles, Torrance, California 90502, USA.
authors.forename: Jie
authors.initials: J
authors.lastname: Wang
authors.forename: Andrew
authors.initials: A
authors.lastname: Lin
authors.forename: Luo
authors.initials: L
authors.lastname: Lu
mesh_terms: D015153:Blotting, Western; D002465:Cell Movement / Q000187:drug effects; D002478:Cells, Cultured; D004789:Enzyme Activation; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D019573:Epithelium, Corneal / Q000187:drug effects* / Q000201:enzymology; D000073864:Histone Deacetylase 6; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology; D006655:Histone Deacetylases / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D009924:Organ Culture Techniques; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000494:pharmacology; D014404:Tubulin / Q000378:metabolism; D014945:Wound Healing / Q000187:drug effects*
chemical_list: D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D014404:Tubulin; C012589:trichostatin A; D004815:Epidermal Growth Factor; C433811:HDAC6 protein, human; D000073864:Histone Deacetylase 6; D006655:Histone Deacetylases