Rho kinases regulate corneal epithelial wound healing.
We have previously shown that Rho small GTPase is required for modulating both cell migration and proliferation through cytoskeleton reorganization and focal adhesion formation in response to wounding. In the present study, we investigated the role of Rho kinases (ROCKs), major effectors of Rho GTPase, in mediating corneal epithelial wound healing. Both ROCK 1 and 2 were expressed and activated in THCE cells, an SV40-immortalized human corneal epithelial cell (HCEC) line, in response to wounding, lysophosphatidic acid, and heparin-binding EGF-like growth factor (HB-EGF) stimulations. The ROCK inhibitor Y-27632 efficiently antagonized ROCK activities without affecting Rho activation in wounded HCECs. Y-27632 promoted basal and HB-EGF-enhanced scratch wound healing and enhanced cell migration and adhesion to matrices, while retarded HB-EGF induced cell proliferation. E-cadherin- and beta-catenin-mediated cell-cell junction and actin cytoskeleton organization were disrupted by Y-27632. Y-27632 impaired the formation and maintenance of tight junction barriers indicated by decreased trans-epithelial resistance and disrupted occludin staining. We conclude that ROCK activities enhance cell proliferation, promote epithelial differentiation, but negatively modulate cell migration and cell adhesion and therefore play a role in regulating corneal epithelial wound healing.
Yin
Jia
J
Department of Ophthalmology, Kresge Eye Institute, Wayne State Univ. School of Medicine, 4717 St. Antoine Blvd, Detroit, MI, 48201, USA.
Yu
Fu-Shin X
FS
D022005:Adherens Junctions / Q000187:drug effects / Q000378:metabolism; D000577:Amides / Q000494:pharmacology; D015153:Blotting, Western; D015820:Cadherins / Q000378:metabolism; D002448:Cell Adhesion / Q000187:drug effects; D002461:Cell Line, Transformed; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D019573:Epithelium, Corneal / Q000166:cytology / Q000378:metabolism / Q000502:physiology; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D066257:Heparin-binding EGF-like Growth Factor; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology; D008246:Lysophospholipids / Q000494:pharmacology; D008565:Membrane Proteins / Q000378:metabolism; D043263:Myosin-Light-Chain Phosphatase / Q000378:metabolism; D062793:Occludin; D010766:Phosphorylation / Q000187:drug effects; D011725:Pyridines / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013314:Stress, Mechanical; D019108:Tight Junctions / Q000187:drug effects / Q000502:physiology; D014945:Wound Healing; D051176:beta Catenin / Q000378:metabolism; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*
D000577:Amides; D015820:Cadherins; D004791:Enzyme Inhibitors; C587883:HBEGF protein, human; D066257:Heparin-binding EGF-like Growth Factor; D036341:Intercellular Signaling Peptides and Proteins; D008246:Lysophospholipids; D008565:Membrane Proteins; C568985:OCLN protein, human; D062793:Occludin; D011725:Pyridines; D051176:beta Catenin; C108830:Y 27632; D054460:rho-Associated Kinases; D048049:Extracellular Signal-Regulated MAP Kinases; D043263:Myosin-Light-Chain Phosphatase; C490372:PPP1R12A protein, human; C032881:lysophosphatidic acid