title: Differentiated thyroid cancer cell invasion is regulated through epidermal growth factor receptor-dependent activation of matrix metalloproteinase (MMP)-2/gelatinase A.
abstract: Mechanisms of invasion in thyroid cancer remain poorly understood. We hypothesized that signaling via the epidermal growth factor receptor (EGFR) stimulates thyroid cancer cell invasion by altering the expression and cleavage of matrix metalloproteinases (MMPs). Papillary and follicular carcinoma cell lines were treated with EGF, the EGFR tyrosine kinase inhibitor AG1478, and the MMP inhibitors GM-6001 and Col-3. Flow cytometry was used to detect EGFR. In vitro invasion assays, gelatin zymography, and quantitative reverse transcription-PCR were used to assess the changes in invasive behavior and MMP expression and activation. All cell lines were found to overexpress functional EGFR. EGF stimulated invasion by thyroid cancer cells up to sevenfold (P<0.0001), a process that was antagonized completely by AG1478 and Col-3, partially by GM-6001, but not by the serine protease inhibitor aprotinin. EGF upregulated expression of MMP-9 (2.64- to 8.89-fold, P<0.0001) and membrane type-1 MMP (MT1-MMP, 1.97- to 2.67-fold, P<0.0001). This effect was blocked completely by AG1478 and partially by Col-3. The activation of MMP-2 paralleled MT1-MMP expression. We demonstrate that MMPs are critical effectors of invasion in the papillary and follicular thyroid cancer cell lines studied. Invasion is regulated by signaling through EGFR, an effect mediated by augmentation of gelatinase expression and activation. MMP inhibitors and growth factor antagonists may be effective tumoristatic agents for the treatment of aggressive thyroid carcinomas.
authors.affiliation: Endocrine Surgery Laboratory, UCSF/Mt. Zion Medical Center, San Francisco, California 94115, USA. myeh@mednet.ucla.edu
authors.forename: Michael W
authors.initials: MW
authors.lastname: Yeh
authors.forename: Jean-Philippe
authors.initials: JP
authors.lastname: Rougier
authors.forename: Jin-Woo
authors.initials: JW
authors.lastname: Park
authors.forename: Quan-Yang
authors.initials: QY
authors.lastname: Duh
authors.forename: Mariwil
authors.initials: M
authors.lastname: Wong
authors.forename: Zena
authors.initials: Z
authors.lastname: Werb
authors.forename: Orlo H
authors.initials: OH
authors.lastname: Clark
mesh_terms: D018263:Adenocarcinoma, Follicular / Q000378:metabolism / Q000473:pathology*; D007611:Aprotinin / Q000494:pharmacology; D015153:Blotting, Western; D002291:Carcinoma, Papillary / Q000378:metabolism / Q000473:pathology*; D002454:Cell Differentiation; D004151:Dipeptides / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004815:Epidermal Growth Factor / Q000494:pharmacology; D066246:ErbB Receptors / Q000235:genetics / Q000378:metabolism*; D005434:Flow Cytometry; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000235:genetics / Q000378:metabolism*; D061965:Matrix Metalloproteinase Inhibitors; D009361:Neoplasm Invasiveness; D011480:Protease Inhibitors / Q000494:pharmacology; D054562:Protein Tyrosine Phosphatase, Non-Receptor Type 1; D017027:Protein Tyrosine Phosphatases / Q000037:antagonists & inhibitors; D011799:Quinazolines; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D013754:Tetracyclines / Q000494:pharmacology; D013964:Thyroid Neoplasms / Q000378:metabolism / Q000473:pathology*; D014407:Tumor Cells, Cultured / Q000378:metabolism; D020032:Tyrphostins / Q000494:pharmacology
chemical_list: D004151:Dipeptides; D004791:Enzyme Inhibitors; D061965:Matrix Metalloproteinase Inhibitors; C078131:N-(2(R)-2-(hydroxamidocarbonylmethyl)-4-methylpentanoyl)-L-tryptophan methylamide; D011480:Protease Inhibitors; D011799:Quinazolines; D015842:Serine Proteinase Inhibitors; D013754:Tetracyclines; D020032:Tyrphostins; C117155:tetracycline CMT-3; C101044:RTKI cpd; D004815:Epidermal Growth Factor; D007611:Aprotinin; D066246:ErbB Receptors; D054562:Protein Tyrosine Phosphatase, Non-Receptor Type 1; D017027:Protein Tyrosine Phosphatases; D020778:Matrix Metalloproteinase 2