title: HaCaT keratinocyte migration is dependent on epidermal growth factor receptor signaling and glycogen synthase kinase-3alpha.
abstract: After epithelial disruption by tissue injury, keratinocytes migrate from the wound edge into a provisional matrix. This process is stimulated by growth factors that signal through epidermal growth factor (EGF) receptor, including EGF, heparin-binding EGF-like growth factor (HB-EGF) and transforming growth factor-alpha (TGF-alpha), and by for example keratinocyte growth factor (KGF) and TGF-beta1 that function through different receptors. We have previously shown that keratinocyte migration induced by EGF or staurosporine is dependent on the activity of glycogen synthase kinase-3 (GSK-3). In the present study, we show that keratinocyte migration induced by TGF-beta1, KGF, EGF, TGF-alpha and staurosporine depends on EGFR signaling, involves autocrine HB-EGF expression and is potently blocked by GSK-3 inhibitors SB-415286 and LiCl. Inhibition of GSK-3 also retards wound reepithelialization in vivo in mice. Moreover, inhibition of GSK-3 activity prevented cell rounding that is an early event in EGFR-mediated keratinocyte migration. Isoform-specific GSK-3alpha and GSK-3beta knockdown and overexpression experiments with siRNAs and adenoviral constructs, respectively, revealed that GSK-3alpha is required for keratinocyte migration, whereas excessive activity of GSK-3beta is inhibitory. Thus, induction of keratinocyte migration is conveyed through EGFR, promoted by endogenous HB-EGF and requires GSK-3alpha activity.
authors.affiliation: University of British Columbia, Faculty of Dentistry, Department of Oral Biological and Medical Sciences, 2199 Wesbrook Mall, Vancouver, Canada BC V6T 1Z3. koivisto@interchange.ubc.ca
authors.forename: Leeni
authors.initials: L
authors.lastname: Koivisto
authors.forename: Guoqiao
authors.initials: G
authors.lastname: Jiang
authors.forename: Lari
authors.initials: L
authors.lastname: Häkkinen
authors.forename: Bosco
authors.initials: B
authors.lastname: Chan
authors.forename: Hannu
authors.initials: H
authors.lastname: Larjava
mesh_terms: D000256:Adenoviridae / Q000235:genetics / Q000378:metabolism; D000627:Aminophenols / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D002460:Cell Line; D002465:Cell Movement; D004815:Epidermal Growth Factor / Q000037:antagonists & inhibitors / Q000378:metabolism; D066246:ErbB Receptors / Q000378:metabolism*; D038362:Glycogen Synthase Kinase 3 / Q000037:antagonists & inhibitors / Q000378:metabolism*; D066257:Heparin-binding EGF-like Growth Factor; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins; D015603:Keratinocytes / Q000166:cytology / Q000201:enzymology / Q000502:physiology*; D018021:Lithium Chloride / Q000378:metabolism / Q000494:pharmacology; D008301:Maleimides / Q000378:metabolism / Q000494:pharmacology; D051379:Mice; D020033:Protein Isoforms / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D051381:Rats; D015398:Signal Transduction; D012867:Skin / Q000166:cytology / Q000378:metabolism; D014162:Transfection; D014945:Wound Healing
chemical_list: C417520:3-(3-chloro-4-hydroxyphenylamino)-4-(4-nitrophenyl)-1H-pyrrole-2,5-dione; D000627:Aminophenols; C587883:HBEGF protein, human; C587884:Hbegf protein, mouse; C587885:Hbegf protein, rat; D066257:Heparin-binding EGF-like Growth Factor; D036341:Intercellular Signaling Peptides and Proteins; D008301:Maleimides; D020033:Protein Isoforms; D034741:RNA, Small Interfering; D004815:Epidermal Growth Factor; D066246:ErbB Receptors; D038362:Glycogen Synthase Kinase 3; C448021:glycogen synthase kinase 3 alpha; D018021:Lithium Chloride