Role of epidermal growth factor and epidermal growth factor receptor on hemidesmosome complex formation and integrin subunit beta4.
Epidermal growth factor (EGF) stimulates integrin beta4 expression and synthesis in corneal epithelium through ligand binding to the EGF receptor, receptor dimerization and activation of the intracellular domain. We hypothesized that inhibition of EGF receptor messenger RNA (mRNA) would block integrin beta4 expression, which is induced by EGF. We also tested the hypothesis that EGF would cause the degradation of hemidesmosomes in control and injured corneal organ cultures. Primary rabbit corneal epithelial cell cultures or corneas were cultured in keratinocyte medium in the presence or absence of an antisense 20-mer phosphorothioate oligonucleotide complementary to the initiation codon region of EGF receptor mRNA. Cells were also cultured in the presence or absence of EGF. Sense and scrambled oligonucleotides similarly modified were used as controls. The concentration of EGF receptor mRNA was semiquantitatively determined by reverse transcriptase/polymerase chain reaction (RT-PCR). We found that transfection did inhibit EGFR expression and migration of epithelial cells and also demonstrated that EGFR mediated expression of integrin beta4 mRNA. Injury induced a decrease in hemidesmosomes that was enhanced with EGF but was not caused by the presence of growth factor in unwounded tissue. These results indicate that injury causes the activation of EGFR but that EGF alone is not responsible for the degradation of hemidesmosomes and that other growth factors play a role in the complex repair of wounds in an avascular tissue.
Song
Q H
QH
Department of Biochemistry, Boston University School of Medicine, MA 02118, USA.
Gong
H
H
Trinkaus-Randall
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V
D000818:Animals; D015152:Blotting, Northern; D002465:Cell Movement / Q000502:physiology; D003896:Desmosomes / Q000502:physiology* / Q000648:ultrastructure; D015536:Down-Regulation; D004815:Epidermal Growth Factor / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D066246:ErbB Receptors / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D005123:Eye / Q000166:cytology / Q000378:metabolism; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D039663:Integrin beta4 / Q000378:metabolism*; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D016376:Oligonucleotides, Antisense; D009924:Organ Culture Techniques; D015347:RNA Probes; D012333:RNA, Messenger / Q000096:biosynthesis / Q000302:isolation & purification; D011817:Rabbits
D039663:Integrin beta4; D016376:Oligonucleotides, Antisense; D015347:RNA Probes; D012333:RNA, Messenger; D004815:Epidermal Growth Factor; D066246:ErbB Receptors