Limits...
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.

Show MeSH

Related in: MedlinePlus

Fibroblast MT1-MMP mediates collagen-invasive activity. (A–C) Wild-type and  fibroblasts were cultured atop type I collagen gels (2.2 mg/ml; top two rows, 2-D) or within three-dimensional collagen gels (2.2 mg/ml; bottom row) in 10% serum and stimulated with a PDGF gradient (10 ng/ml) for 6 d in the absence or presence of TIMP-2. The arrow to the left of the panels marks the surface of the collagen monolayer or the edge of the embedded island of fibroblasts. Arrows within the gel indicate positions of invading fibroblasts. Bars, 100 μm. Invasion (B) and collagen degradation (C) were quantified in three-dimensional collagen gels in the presence of 10% wild-type serum save for experiments with MMP-2−/−, MMP-9−/−, or TIMP-2−/− fibroblasts, where  serum was isolated from each of the respective mouse strains. Results are expressed as the mean ± 1 SEM of three or more experiments.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2172570&req=5

fig4: Fibroblast MT1-MMP mediates collagen-invasive activity. (A–C) Wild-type and fibroblasts were cultured atop type I collagen gels (2.2 mg/ml; top two rows, 2-D) or within three-dimensional collagen gels (2.2 mg/ml; bottom row) in 10% serum and stimulated with a PDGF gradient (10 ng/ml) for 6 d in the absence or presence of TIMP-2. The arrow to the left of the panels marks the surface of the collagen monolayer or the edge of the embedded island of fibroblasts. Arrows within the gel indicate positions of invading fibroblasts. Bars, 100 μm. Invasion (B) and collagen degradation (C) were quantified in three-dimensional collagen gels in the presence of 10% wild-type serum save for experiments with MMP-2−/−, MMP-9−/−, or TIMP-2−/− fibroblasts, where serum was isolated from each of the respective mouse strains. Results are expressed as the mean ± 1 SEM of three or more experiments.

Mentions: Although fibroblasts and tumor cells both use MT1-MMP as a pericellular collagenolysin, the invasive phenotype displayed by cells negotiating three-dimensional ECM barriers is a more complex form of cellular behavior and conceivably requires complementation by other proteolytic systems (Murphy and Gavrilovic, 1999). Hence, monolayers of wild-type and fibroblasts were established atop three-dimensional collagen gels and the cells exposed to a chemotactic gradient of PDGF. Under these conditions, MMP-13−/−, MMP-8−/−, MMP-2−/−, TIMP-2−/−, MMP-3−/−, and MMP-9−/− fibroblasts each invade the collagen gels comparably to wild-type cells while degrading similar amounts of collagen (Fig. 4, A–C). Likewise, though plasmin can activate secreted MMPs, fibroblasts isolated from plasminogen- mice and suspended in plasminogen- sera display an invasive phenotype indistinguishable from controls (i.e., 98 ± 10% of control; n = 3). Recent papers have promoted a potential role for the transmembrane serine prolyl peptidase, fibroblast activation protein/seprase, in collagen remodeling events (Ghersi et al., 2002), but the fibroblasts also retained full invasive activity (i.e., and 95 ± 6% of control; n = 3). In contrast, MT1-MMP−/− fibroblasts are unable to express an invasive phenotype over the 6-d culture period (Fig. 4, A–C). Though cell behavior can be altered when cells are embedded within, as opposed to cultured atop, a three-dimensional matrix (Cukierman et al., 2002, Hotary et al., 2003), MT1-MMP−/− cells sustain a similar, and complete, deficiency in three-dimensional invasive behavior relative to wild-type fibroblasts (Fig. 4 A). However, under either two- or three-dimensional conditions, the MT1-MMP−/− invasion- phenotype is rescued when cells are transfected with an expression vector encoding wild-type (Fig. 4 A), but not either soluble ΔMT1-MMP, inactive MT1-MMP E240→A, mColA, human MMP-1RXKR, human MMP-13RXKR, or human MMP-2RXKR (not depicted). However, defects in MT1-MMP−/− fibroblasts are confined to the type I collagen substrate as the cells migrate across collagen-coated surfaces comparably to controls, exert a normal collagen-contractile response (Fig. S4, available at http://www.jcb.org/cgi/content/full/jcb.200408028/DC1), and, as reported previously, invade three-dimensional matrices of cross-linked fibrin via an MT3-MMP–dependent process (Hotary et al., 2002).


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)

Fibroblast MT1-MMP mediates collagen-invasive activity. (A–C) Wild-type and  fibroblasts were cultured atop type I collagen gels (2.2 mg/ml; top two rows, 2-D) or within three-dimensional collagen gels (2.2 mg/ml; bottom row) in 10% serum and stimulated with a PDGF gradient (10 ng/ml) for 6 d in the absence or presence of TIMP-2. The arrow to the left of the panels marks the surface of the collagen monolayer or the edge of the embedded island of fibroblasts. Arrows within the gel indicate positions of invading fibroblasts. Bars, 100 μm. Invasion (B) and collagen degradation (C) were quantified in three-dimensional collagen gels in the presence of 10% wild-type serum save for experiments with MMP-2−/−, MMP-9−/−, or TIMP-2−/− fibroblasts, where  serum was isolated from each of the respective mouse strains. Results are expressed as the mean ± 1 SEM of three or more experiments.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2172570&req=5

fig4: Fibroblast MT1-MMP mediates collagen-invasive activity. (A–C) Wild-type and fibroblasts were cultured atop type I collagen gels (2.2 mg/ml; top two rows, 2-D) or within three-dimensional collagen gels (2.2 mg/ml; bottom row) in 10% serum and stimulated with a PDGF gradient (10 ng/ml) for 6 d in the absence or presence of TIMP-2. The arrow to the left of the panels marks the surface of the collagen monolayer or the edge of the embedded island of fibroblasts. Arrows within the gel indicate positions of invading fibroblasts. Bars, 100 μm. Invasion (B) and collagen degradation (C) were quantified in three-dimensional collagen gels in the presence of 10% wild-type serum save for experiments with MMP-2−/−, MMP-9−/−, or TIMP-2−/− fibroblasts, where serum was isolated from each of the respective mouse strains. Results are expressed as the mean ± 1 SEM of three or more experiments.
Mentions: Although fibroblasts and tumor cells both use MT1-MMP as a pericellular collagenolysin, the invasive phenotype displayed by cells negotiating three-dimensional ECM barriers is a more complex form of cellular behavior and conceivably requires complementation by other proteolytic systems (Murphy and Gavrilovic, 1999). Hence, monolayers of wild-type and fibroblasts were established atop three-dimensional collagen gels and the cells exposed to a chemotactic gradient of PDGF. Under these conditions, MMP-13−/−, MMP-8−/−, MMP-2−/−, TIMP-2−/−, MMP-3−/−, and MMP-9−/− fibroblasts each invade the collagen gels comparably to wild-type cells while degrading similar amounts of collagen (Fig. 4, A–C). Likewise, though plasmin can activate secreted MMPs, fibroblasts isolated from plasminogen- mice and suspended in plasminogen- sera display an invasive phenotype indistinguishable from controls (i.e., 98 ± 10% of control; n = 3). Recent papers have promoted a potential role for the transmembrane serine prolyl peptidase, fibroblast activation protein/seprase, in collagen remodeling events (Ghersi et al., 2002), but the fibroblasts also retained full invasive activity (i.e., and 95 ± 6% of control; n = 3). In contrast, MT1-MMP−/− fibroblasts are unable to express an invasive phenotype over the 6-d culture period (Fig. 4, A–C). Though cell behavior can be altered when cells are embedded within, as opposed to cultured atop, a three-dimensional matrix (Cukierman et al., 2002, Hotary et al., 2003), MT1-MMP−/− cells sustain a similar, and complete, deficiency in three-dimensional invasive behavior relative to wild-type fibroblasts (Fig. 4 A). However, under either two- or three-dimensional conditions, the MT1-MMP−/− invasion- phenotype is rescued when cells are transfected with an expression vector encoding wild-type (Fig. 4 A), but not either soluble ΔMT1-MMP, inactive MT1-MMP E240→A, mColA, human MMP-1RXKR, human MMP-13RXKR, or human MMP-2RXKR (not depicted). However, defects in MT1-MMP−/− fibroblasts are confined to the type I collagen substrate as the cells migrate across collagen-coated surfaces comparably to controls, exert a normal collagen-contractile response (Fig. S4, available at http://www.jcb.org/cgi/content/full/jcb.200408028/DC1), and, as reported previously, invade three-dimensional matrices of cross-linked fibrin via an MT3-MMP–dependent process (Hotary et al., 2002).

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