Tumor necrosis factor-alpha-stimulated membrane type 1-matrix metalloproteinase production is modulated by epidermal growth factor receptor signaling in human gingival fibroblasts.
OBJECTIVE
Membrane type 1-matrix metalloproteinase (MT1-MMP) is a collagenolytic enzyme involved in connective tissue remodeling. In periodontal tissues, either cytokines or growth factors regulate the production of proteolytic enzymes. Mice deficient in epidermal growth factor receptor (EGFR) show a reduced expression of MT1-MMP, suggesting that this receptor may play an important role in MT1-MMP production. The present study evaluated the role of the inflammatory cytokine tumor necrosis factor-alpha (TNF-alpha) and EGFR in the production of MT1-MMP in gingival fibroblasts.


METHODS
Primary cultures of human gingival fibroblasts were cultured over plastic or a type I collagen matrix and stimulated with TNF-alpha and EGF. A selective EGFR inhibitor (AG1478) was used to interfere with this signaling pathway. Production of MT1-MMP and activation of proMMP-2 were studied using Western blot and gelatin zymography, respectively. Activation of EGFR signaling was assessed through immunoprecipitation and Western blot. Expression of EGFR ligands was determined through reverse transcriptase-polymerase chain reaction.


RESULTS
Treatment of gingival fibroblasts cultured over a collagen matrix with TNF-alpha stimulated proMMP-2 activation and MT1-MMP production. However, after using AG1478, both responses were inhibited. Tumor necrosis factor-alpha induced EGFR transactivation and stimulated the expression of the mRNA for the EGFR ligands heparin binding-epidermal growth factor (HB-EGF) and transforming growth factor-alpha (TGF-alpha).


CONCLUSIONS
The present study shows that TNF-alpha may stimulate MT1-MMP production through transactivation of EGFR. Tumor necrosis factor-alpha may also modulate the expression of the EGFR ligands TGF-alpha and HB-EGF. Production of MT1-MMP by TNF-alpha requires interaction with EGFR, suggesting that tissue remodeling is controlled by cross-communication between diverse signaling pathways in gingival fibroblasts.
Smith
P C
PC
Faculty of Odontology, University of Chile, Santiago, Chile. patricio.smith@gmail.com
Guerrero
J
J
Tobar
N
N
Cáceres
M
M
González
M J
MJ
Martínez
J
J
D002478:Cells, Cultured; D024042:Collagen Type I / Q000494:pharmacology; D003470:Culture Media; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004792:Enzyme Precursors / Q000187:drug effects; D004815:Epidermal Growth Factor / Q000494:pharmacology; D066246:ErbB Receptors / Q000037:antagonists & inhibitors / Q000187:drug effects*; D005347:Fibroblasts / Q000187:drug effects* / Q000201:enzymology; D018093:Gelatinases / Q000187:drug effects; D005881:Gingiva / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D006493:Heparin / Q000032:analysis; D066257:Heparin-binding EGF-like Growth Factor; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000032:analysis; D008024:Ligands; D053511:Matrix Metalloproteinase 14 / Q000187:drug effects*; D010766:Phosphorylation; D017027:Protein Tyrosine Phosphatases / Q000037:antagonists & inhibitors; D011799:Quinazolines; D011956:Receptors, Cell Surface / Q000032:analysis; D015398:Signal Transduction / Q000187:drug effects*; D015533:Transcriptional Activation / Q000187:drug effects; D016211:Transforming Growth Factor alpha / Q000187:drug effects; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*; D020032:Tyrphostins / Q000494:pharmacology
D024042:Collagen Type I; D003470:Culture Media; D004791:Enzyme Inhibitors; D004792:Enzyme Precursors; C587883:HBEGF protein, human; C587884:Hbegf protein, mouse; D066257:Heparin-binding EGF-like Growth Factor; D036341:Intercellular Signaling Peptides and Proteins; D008024:Ligands; D011799:Quinazolines; D011956:Receptors, Cell Surface; D016211:Transforming Growth Factor alpha; D014409:Tumor Necrosis Factor-alpha; D020032:Tyrphostins; C101044:RTKI cpd; D004815:Epidermal Growth Factor; D006493:Heparin; D066246:ErbB Receptors; D017027:Protein Tyrosine Phosphatases; D018093:Gelatinases; C069860:progelatinase; D053511:Matrix Metalloproteinase 14