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Tumor cell traffic through the extracellular matrix is controlled by the membrane-anchored collagenase MT1-MMP.

Sabeh F, Ota I, Holmbeck K, Birkedal-Hansen H, Soloway P, Balbin M, Lopez-Otin C, Shapiro S, Inada M, Krane S, Allen E, Chung D, Weiss SJ - J. Cell Biol. (2004)

Bottom Line: As cancer cells traverse collagen-rich extracellular matrix (ECM) barriers and intravasate, they adopt a fibroblast-like phenotype and engage undefined proteolytic cascades that mediate invasive activity.Herein, we find that fibroblasts and cancer cells express an indistinguishable pericellular collagenolytic activity that allows them to traverse the ECM.Thus, MT1-MMP serves as the major cell-associated proteinase necessary to confer normal or neoplastic cells with invasive activity.

View Article: PubMed Central - PubMed

Affiliation: Division of Molecular Medicine and Genetics, Department of Internal Medicine, University of Michigan, Ann Arbor, MI 48109, USA.

ABSTRACT
As cancer cells traverse collagen-rich extracellular matrix (ECM) barriers and intravasate, they adopt a fibroblast-like phenotype and engage undefined proteolytic cascades that mediate invasive activity. Herein, we find that fibroblasts and cancer cells express an indistinguishable pericellular collagenolytic activity that allows them to traverse the ECM. Using fibroblasts isolated from gene-targeted mice, a matrix metalloproteinase (MMP)-dependent activity is identified that drives invasion independently of plasminogen, the gelatinase A/TIMP-2 axis, gelatinase B, collagenase-3, collagenase-2, or stromelysin-1. In contrast, deleting or suppressing expression of the membrane-tethered MMP, MT1-MMP, in fibroblasts or tumor cells results in a loss of collagenolytic and invasive activity in vitro or in vivo. Thus, MT1-MMP serves as the major cell-associated proteinase necessary to confer normal or neoplastic cells with invasive activity.

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CAM invasion by wild-type and MMP- fibroblasts. (A) Fibroblasts from wild-type, MMP-deficient, or FAP- mice were labeled with fluorescent microbeads (green) and seeded atop the CAM of 11-d-old chicks and cultured for 3 d. Fluorescent micrographs of CAM cross sections demonstrate the ability of mouse fibroblasts to penetrate the CAM surface (DAPI-stained, blue nuclei of chick cells at the CAM surface are highlighted by arrows). The MT1-MMP−/− defect was reversed by transient transfection of MT1-MMP−/− cells with MT1-MMP. Results are representative of four performed. Bar, 100 μm. (B) Invasion was quantified as the percentage of wild-type or  fibroblasts that traversed the CAM surface and the mean depth of invasion of the leading front of three or more fibroblasts. Results are expressed as the mean ± 1 SD of at least three experiments.
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fig5: CAM invasion by wild-type and MMP- fibroblasts. (A) Fibroblasts from wild-type, MMP-deficient, or FAP- mice were labeled with fluorescent microbeads (green) and seeded atop the CAM of 11-d-old chicks and cultured for 3 d. Fluorescent micrographs of CAM cross sections demonstrate the ability of mouse fibroblasts to penetrate the CAM surface (DAPI-stained, blue nuclei of chick cells at the CAM surface are highlighted by arrows). The MT1-MMP−/− defect was reversed by transient transfection of MT1-MMP−/− cells with MT1-MMP. Results are representative of four performed. Bar, 100 μm. (B) Invasion was quantified as the percentage of wild-type or fibroblasts that traversed the CAM surface and the mean depth of invasion of the leading front of three or more fibroblasts. Results are expressed as the mean ± 1 SD of at least three experiments.

Mentions: Although fibroblasts use MT1-MMP to cross barriers of type I collagen assembled in vitro, the extracellular matrix in vivo is a more complex mix of collagens, glycosaminoglycans, glycoproteins, and peptidoglycans (Hay, 1991). To determine whether or not MT1-MMP likewise regulates tissue-invasive activity in vivo, wild-type or fibroblasts labeled with fluorescent nanobeads were applied to the surface of the chicken chorioallantoic membrane (CAM), a type I collagen-rich ECM barrier commonly used to study invasive processes (Kim et al., 1998). As reported previously (Armstrong et al., 1982), wild-type fibroblasts readily invade the CAM in the course of a 3-d culture period (Fig. 5 A). Though the pattern of distribution of the invading fibroblasts is variable between tissue sections and individual chick CAMs (unpublished data), both the percentage of fibroblasts that cross the CAM surface and the depth of the leading front of invasion is highly reproducible. By either of these criteria, neither MMP-13−/−, MMP-8−/−, MMP-2−/−, MMP-3−/−, MMP-9−/−, plasminogen−/−, nor fibroblast activation protein−/− fibroblasts display discernible defects in invasive activity (Fig. 5, A and B). In contrast, MT1-MMP- fibroblasts are completely unable to penetrate the CAM surface unless the cells are transfected with wild-type, but not a MT1-MMP E240→A, expression vector (Fig. 5, B and C).


Tumor cell traffic through the extracellular matrix is controlled by the membrane-anchored collagenase MT1-MMP.

Sabeh F, Ota I, Holmbeck K, Birkedal-Hansen H, Soloway P, Balbin M, Lopez-Otin C, Shapiro S, Inada M, Krane S, Allen E, Chung D, Weiss SJ - J. Cell Biol. (2004)

CAM invasion by wild-type and MMP- fibroblasts. (A) Fibroblasts from wild-type, MMP-deficient, or FAP- mice were labeled with fluorescent microbeads (green) and seeded atop the CAM of 11-d-old chicks and cultured for 3 d. Fluorescent micrographs of CAM cross sections demonstrate the ability of mouse fibroblasts to penetrate the CAM surface (DAPI-stained, blue nuclei of chick cells at the CAM surface are highlighted by arrows). The MT1-MMP−/− defect was reversed by transient transfection of MT1-MMP−/− cells with MT1-MMP. Results are representative of four performed. Bar, 100 μm. (B) Invasion was quantified as the percentage of wild-type or  fibroblasts that traversed the CAM surface and the mean depth of invasion of the leading front of three or more fibroblasts. Results are expressed as the mean ± 1 SD of at least three experiments.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2172570&req=5

fig5: CAM invasion by wild-type and MMP- fibroblasts. (A) Fibroblasts from wild-type, MMP-deficient, or FAP- mice were labeled with fluorescent microbeads (green) and seeded atop the CAM of 11-d-old chicks and cultured for 3 d. Fluorescent micrographs of CAM cross sections demonstrate the ability of mouse fibroblasts to penetrate the CAM surface (DAPI-stained, blue nuclei of chick cells at the CAM surface are highlighted by arrows). The MT1-MMP−/− defect was reversed by transient transfection of MT1-MMP−/− cells with MT1-MMP. Results are representative of four performed. Bar, 100 μm. (B) Invasion was quantified as the percentage of wild-type or fibroblasts that traversed the CAM surface and the mean depth of invasion of the leading front of three or more fibroblasts. Results are expressed as the mean ± 1 SD of at least three experiments.
Mentions: Although fibroblasts use MT1-MMP to cross barriers of type I collagen assembled in vitro, the extracellular matrix in vivo is a more complex mix of collagens, glycosaminoglycans, glycoproteins, and peptidoglycans (Hay, 1991). To determine whether or not MT1-MMP likewise regulates tissue-invasive activity in vivo, wild-type or fibroblasts labeled with fluorescent nanobeads were applied to the surface of the chicken chorioallantoic membrane (CAM), a type I collagen-rich ECM barrier commonly used to study invasive processes (Kim et al., 1998). As reported previously (Armstrong et al., 1982), wild-type fibroblasts readily invade the CAM in the course of a 3-d culture period (Fig. 5 A). Though the pattern of distribution of the invading fibroblasts is variable between tissue sections and individual chick CAMs (unpublished data), both the percentage of fibroblasts that cross the CAM surface and the depth of the leading front of invasion is highly reproducible. By either of these criteria, neither MMP-13−/−, MMP-8−/−, MMP-2−/−, MMP-3−/−, MMP-9−/−, plasminogen−/−, nor fibroblast activation protein−/− fibroblasts display discernible defects in invasive activity (Fig. 5, A and B). In contrast, MT1-MMP- fibroblasts are completely unable to penetrate the CAM surface unless the cells are transfected with wild-type, but not a MT1-MMP E240→A, expression vector (Fig. 5, B and C).

Bottom Line: As cancer cells traverse collagen-rich extracellular matrix (ECM) barriers and intravasate, they adopt a fibroblast-like phenotype and engage undefined proteolytic cascades that mediate invasive activity.Herein, we find that fibroblasts and cancer cells express an indistinguishable pericellular collagenolytic activity that allows them to traverse the ECM.Thus, MT1-MMP serves as the major cell-associated proteinase necessary to confer normal or neoplastic cells with invasive activity.

View Article: PubMed Central - PubMed

Affiliation: Division of Molecular Medicine and Genetics, Department of Internal Medicine, University of Michigan, Ann Arbor, MI 48109, USA.

ABSTRACT
As cancer cells traverse collagen-rich extracellular matrix (ECM) barriers and intravasate, they adopt a fibroblast-like phenotype and engage undefined proteolytic cascades that mediate invasive activity. Herein, we find that fibroblasts and cancer cells express an indistinguishable pericellular collagenolytic activity that allows them to traverse the ECM. Using fibroblasts isolated from gene-targeted mice, a matrix metalloproteinase (MMP)-dependent activity is identified that drives invasion independently of plasminogen, the gelatinase A/TIMP-2 axis, gelatinase B, collagenase-3, collagenase-2, or stromelysin-1. In contrast, deleting or suppressing expression of the membrane-tethered MMP, MT1-MMP, in fibroblasts or tumor cells results in a loss of collagenolytic and invasive activity in vitro or in vivo. Thus, MT1-MMP serves as the major cell-associated proteinase necessary to confer normal or neoplastic cells with invasive activity.

Show MeSH
Related in: MedlinePlus