Glycogen synthase kinase-3 acts upstream of ADP-ribosylation factor 6 and Rac1 to regulate epithelial cell migration

Exp Cell Res. 2006 May 15;312(9):1514-25. doi: 10.1016/j.yexcr.2006.01.018. Epub 2006 Mar 9.


Cell sheet movement during epithelial wound closure is a complex process involving collective cell migration. We have found that glycogen synthase kinase-3 (GSK-3) activity is required for membrane protrusion and crawling of cells at the wound edge and those behind it in wounded Madin-Darby canine kidney (MDCK) epithelial cell monolayers. RNA interference-based silencing of GSK-3alpha and GSK-3beta expression also results in slowed cell sheet migration, with the effect being more pronounced with knockdown of GSK-3beta. Both GSK-3alpha and GSK-3beta are in activated states during the most active phase of cell migration. In addition to having a positive control or permissive, rather than negative, function in MDCK cell migration, GSK-3 appears to act upstream of the small GTPases ADP-ribosylation factor 6 (ARF6) and Rac1. Expression of constitutively active ARF6 restores a protrusive, migratory phenotype in cells treated with GSK-3 inhibitors. It does not, however, restore to normal levels the directional polarization of cells behind the wound edge toward the wound area, implying the existence of a separate ARF6-independent branch of the GSK-3 pathway that regulates proper wound-directed polarization of these cells. Finally, inhibition of GSK-3 also strongly reduces activation of Rac1 and cell scatter in response to hepatocyte growth factor/scatter factor, which triggers dispersal and migration of cells in monolayer culture as fibroblast-like individual cells, a mode of epithelial cell motility distinct from the collective migration of wound closure.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ADP-Ribosylation Factor 6
  • ADP-Ribosylation Factors / metabolism*
  • Aminophenols / pharmacology
  • Androstadienes / pharmacology
  • Animals
  • Cell Line
  • Cell Movement / drug effects
  • Cell Movement / genetics
  • Cell Movement / physiology*
  • Cell Surface Extensions / drug effects
  • Cell Survival / drug effects
  • Dogs
  • Doxycycline / pharmacology
  • Enzyme Inhibitors / pharmacology
  • Epithelial Cells / cytology
  • Epithelial Cells / drug effects
  • Epithelial Cells / metabolism*
  • Gene Expression / drug effects
  • Glycogen Synthase Kinase 3 / antagonists & inhibitors
  • Glycogen Synthase Kinase 3 / genetics
  • Glycogen Synthase Kinase 3 / metabolism*
  • Glycogen Synthase Kinase 3 beta
  • Hepatocyte Growth Factor / pharmacology
  • Maleimides / pharmacology
  • Models, Biological
  • Phosphoinositide-3 Kinase Inhibitors
  • Phosphorylation / drug effects
  • RNA, Small Interfering / genetics
  • Signal Transduction / drug effects
  • Wortmannin
  • rac1 GTP-Binding Protein / metabolism*


  • 3-(3-chloro-4-hydroxyphenylamino)-4-(4-nitrophenyl)-1H-pyrrole-2,5-dione
  • ADP-Ribosylation Factor 6
  • Aminophenols
  • Androstadienes
  • Enzyme Inhibitors
  • Maleimides
  • Phosphoinositide-3 Kinase Inhibitors
  • RNA, Small Interfering
  • Hepatocyte Growth Factor
  • Glycogen Synthase Kinase 3 beta
  • Glycogen Synthase Kinase 3
  • glycogen synthase kinase 3 alpha
  • ADP-Ribosylation Factors
  • rac1 GTP-Binding Protein
  • Doxycycline
  • Wortmannin