The effect of epidermal growth factor on matrix metalloproteinases and tissue inhibitors of metalloproteinase gene expression in cultured human gingival fibroblasts.
OBJECTIVE
Matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) play a role in the breakdown of the extracellular matrix during normal physiological processes, and in pathological processes, including periodontitis. The aim of this study was to evaluate the effect of epidermal growth factor (EGF) on the expression of MMPs and TIMPs in cultured human gingival fibroblasts.


METHODS
Fibroblasts were stimulated with 10(-3), 10(-6) or 10(-12)M EGF for 24h; untreated fibroblasts served as controls. Alterations in the expression of MMP-1, 2, 3, 7, 11, TIMP-1 and 2 were evaluated using real-time PCR and Western blotting. beta-Actin expression was used as a reference to normalize gene expression.


RESULTS
Increased MMP-1, 3, 7 and 11 expressions were observed at all EGF concentrations (p<0.05). At the lowest EGF concentration, MMP-1, 3 and 7 presented the lowest expression and MMP-11 presented the greatest expression; at higher EGF concentrations, MMP-1, 3 and 7 presented greater up-regulation, and MMP-11 lower up-regulation (p<0.05). Protein expression was similarly regulated by EGF: increased up-regulation of MMP-1, 3 and 7 was observed with increasing EGF concentrations, except for MMP-11 that exhibited greater up-regulation at the lower EGF concentration. The gene expression of MMP-2, TIMP-1 and 2 was not affected by EGF (p<0.05).


CONCLUSIONS
We conclude that EGF regulates expression for MMP-1, 3, 7 and 11 in a dose-dependent manner, suggesting that EGF may play a role in periodontal destruction and wound repair.
Cury
Patricia R
PR
Department of Oral Pathology, University of São Paulo, School of Dentistry, São Paulo, Brazil. patcury@yahoo.com
de Araújo
Vera C
VC
Canavez
Flávio
F
Furuse
Cristiane
C
Leite
Kátia R M
KR
de Araújo
Ney S
NS
D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D005347:Fibroblasts / Q000187:drug effects* / Q000201:enzymology; D005786:Gene Expression Regulation / Q000187:drug effects; D005881:Gingiva / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D006801:Humans; D020781:Matrix Metalloproteinase 1 / Q000032:analysis / Q000187:drug effects; D053507:Matrix Metalloproteinase 11 / Q000032:analysis / Q000187:drug effects; D020778:Matrix Metalloproteinase 2 / Q000032:analysis / Q000187:drug effects; D019278:Matrix Metalloproteinase 3 / Q000032:analysis / Q000187:drug effects; D020783:Matrix Metalloproteinase 7 / Q000032:analysis / Q000187:drug effects; D020782:Matrix Metalloproteinases / Q000032:analysis / Q000187:drug effects*; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000032:analysis / Q000187:drug effects; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000032:analysis / Q000187:drug effects; D019714:Tissue Inhibitor of Metalloproteinases / Q000032:analysis / Q000187:drug effects*; D015854:Up-Regulation / Q000187:drug effects
D019715:Tissue Inhibitor of Metalloproteinase-1; D019714:Tissue Inhibitor of Metalloproteinases; D019716:Tissue Inhibitor of Metalloproteinase-2; D004815:Epidermal Growth Factor; C511771:MMP11 protein, human; D053507:Matrix Metalloproteinase 11; D020782:Matrix Metalloproteinases; C511768:MMP3 protein, human; D019278:Matrix Metalloproteinase 3; C060932:MMP7 protein, human; D020783:Matrix Metalloproteinase 7; C522361:MMP2 protein, human; D020778:Matrix Metalloproteinase 2; C522360:MMP1 protein, human; D020781:Matrix Metalloproteinase 1