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L. Lu; Effect of EGF-induced Deacetylation on Corneal Epithelial Wound Healing. Invest. Ophthalmol. Vis. Sci. 2009;50(13):6295.
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Epidermal Growth factor (EGF) plays important roles in regulating corneal epithelial proliferation/differentiation during wound healing. The purpose of the study is to investigate effects of EGF-induced deacetylation on corneal epithelial migration and wound healing.
Human and rabbit corneal epithelial cells (HCE and RCE cells) were cultured in DMEM/F12 medium contained 10% FBS in 37 °C incubator supplied with 5% CO2. Northern blot and Western analysis were used to determine target gene and protein expressions. Activity of HDAC6 was suppressed by trichostatin A (TSA) and by siRNA specific to HDAC6 mRNA. Corneal epithelial cell proliferation was measured by corneal epithelial migration and debridement wound healing model.
We found EGF-stimulated corneal epithelial cell migration and wound healing through enhancing HDAC6 activity and deacetylation of -tubulin. We report in the study: 1) EGF induced an increase in HDAC6 activity and enhanced decetylation of -tubulin; 2) EGF-induced HCE cell migration was suppressed by TSA-induced inhibition of HDAC6 activity; 3) knockdown of HDAC6 mRNA with specific siRNA effectively abolished EGF-induced deacetylation of -tubulin and significantly inhibited HCE cell migration; 4) inhibition of Erk signaling pathway with PD98059 suppressed EGF-induced HDAC6 activation; and 5) suppression of HDAC6 activity with TSA significantly delayed EGF-induced wound healing in epithelial debrided corneas.
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