| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
From the Department of Cellular Biology and Anatomy, Medical College of Georgia, Augusta, Georgia.
PURPOSE. Human corneal epithelial cells (HCECs) were functionally depleted of erbB2 to elucidate its role in epidermal growth factor (EGF) receptor (EGFR) activation-dependent cell migration.
METHODS. The retrovirus pBabe-5R, which encodes an erbB2 single-chain antibody with an endoplasmic reticulum (ER)–targeting sequence, and control pBabe-puro were used to infect THCE cells (an SV40-immortalized HCEC line). Several cell lines expressing 5R were selected along with a pBabe-puro control line. The depletion of erbB2 was verified by cell surface biotinylation of proteins, followed by streptavidin precipitation and subsequent detection of erbB2 by immunoblot analysis. Activation of erbBs was analyzed by immunoprecipitation using the phosphotyrosine antibody pY20, followed by Western blot analysis with erbB1 or erbB2 antibodies. Phosphorylation of extracellular signal-regulated kinase (ERK) and phosphatidylinositol 3'-kinase (PI3K) was analyzed by Western blot with antibodies specific to phosphorylated proteins. Effects of erbB2 depletion on heparin-binding EGF-like growth factor (HB-EGF)–induced cell migration were determined by Boyden chamber migration assay and by scratch wound assay.
RESULTS. Wounding induced erbB2 tyrosine phosphorylation. Expression of 5R encoding an erbB2 single-chain antibody with an endoplasmic reticulum–targeting sequence depleted the cell surface expression of erbB2 in HCECs. Wounding resulted in a rapid increase in the phosphorylation of erbB1 in both 5R-expressing and control cells, whereas wound-induced erbB2 phosphorylation in 5R-expressing cells was not detectable. Depletion of functional erbB2 attenuated the healing of scratch wounds in the presence of HB-EGF and impaired both chemotactic migration stimulated by HB-EGF and haptotactic migration toward a fibronectin-collagen I (3:1; FNC) coating mix. Expression of 5R affected both the intensity and the duration of wound-induced, EGFR-elicited ERK and PI3K activation. Inhibition of ERK and PI3K pathways in cultured porcine corneas impaired ex vivo epithelial wound healing.
CONCLUSIONS. ErbB2 serves as a critical component that couples erbB receptor tyrosine kinase to the migration machinery of corneal epithelial cells.
This article has been cited by other articles:
|
|
 |
|
  L. P. Desai, S. R. White, and C. M. Waters Cyclic Mechanical Stretch Decreases Cell Migration by Inhibiting Phosphatidylinositol 3-Kinase- and Focal Adhesion Kinase-mediated JNK1 Activation J. Biol. Chem., February 12, 2010; 285(7): 4511 - 4519. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K.-P. Xu, Y. Li, A. V. Ljubimov, and F.-S. X. Yu High Glucose Suppresses Epidermal Growth Factor Receptor/Phosphatidylinositol 3-Kinase/Akt Signaling Pathway and Attenuates Corneal Epithelial Wound Healing Diabetes, May 1, 2009; 58(5): 1077 - 1085. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Yin and F.-S. X. Yu ERK1/2 Mediate Wounding- and G-protein-Coupled Receptor Ligands-Induced EGFR Activation via Regulating ADAM17 and HB-EGF Shedding Invest. Ophthalmol. Vis. Sci., January 1, 2009; 50(1): 132 - 139. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Y. Ghoghawala, M. J. Mannis, C. E. Pullar, M. I. Rosenblatt, and R. R. Isseroff {beta}2-Adrenergic Receptor Signaling Mediates Corneal Epithelial Wound Repair Invest. Ophthalmol. Vis. Sci., May 1, 2008; 49(5): 1857 - 1863. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Yin, J. Lu, and F.-S. X. Yu Role of Small GTPase Rho in Regulating Corneal Epithelial Wound Healing Invest. Ophthalmol. Vis. Sci., March 1, 2008; 49(3): 900 - 909. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P Specenier, C Koppen, and J. Vermorken Diffuse punctate keratitis in a patient treated with cetuximab as monotherapy Ann. Onc., May 1, 2007; 18(5): 961 - 962. [Full Text] [PDF]
|
|
|
|
 |
|
 J. Yin, K. Xu, J. Zhang, A. Kumar, and F.-S. X. Yu Wound-induced ATP release and EGF receptor activation in epithelial cells J. Cell Sci., March 1, 2007; 120(5): 815 - 825. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K.-P. Xu, J. Yin, and F.-S. X. Yu Lysophosphatidic Acid Promoting Corneal Epithelial Wound Healing by Transactivation of Epidermal Growth Factor Receptor Invest. Ophthalmol. Vis. Sci., February 1, 2007; 48(2): 636 - 643. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. H. Levin and A. S. Verkman Aquaporin-3-Dependent Cell Migration and Proliferation during Corneal Re-epithelialization. Invest. Ophthalmol. Vis. Sci., October 1, 2006; 47(10): 4365 - 4372. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. E. Pullar, A. Rizzo, and R. R. Isseroff beta-Adrenergic Receptor Antagonists Accelerate Skin Wound Healing: EVIDENCE FOR A CATECHOLAMINE SYNTHESIS NETWORK IN THE EPIDERMIS J. Biol. Chem., July 28, 2006; 281(30): 21225 - 21235. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K.-P. Xu, J. Yin, and F.-S. X. Yu SRC-family tyrosine kinases in wound- and ligand-induced epidermal growth factor receptor activation in human corneal epithelial cells. Invest. Ophthalmol. Vis. Sci., July 1, 2006; 47(7): 2832 - 2839. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T.-H. Huang and S. L. Morrison A Trimeric Anti-HER2/neu ScFv and Tumor Necrosis Factor-{alpha} Fusion Protein Induces HER2/neu Signaling and Facilitates Repair of Injured Epithelia J. Pharmacol. Exp. Ther., March 1, 2006; 316(3): 983 - 991. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
