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

Collagen-invasive and -degradative activities of fibroblasts and tumor cells. (A) Transmission electron micrographs of fibroblast monolayers cultured atop three-dimensional collagen gels (2.2 mg/ml) with PDGF-BB (10 ng/ml) in the presence of 10% serum at the start of the culture period (0 d) or after 6 d in either the absence or presence of 5 μM BB-94. Zones of collagen clearing are observed in areas surrounding invading cells (middle, arrows). Double-headed arrow indicates the position of the underlying gel. Bar, 10 μm. (B) Laser confocal micrographs of rhodamine-labeled collagen gels (2.2 mg/ml) incubated with fibroblasts and PDGF in 10% serum for 0 or 6 d in the absence or presence of BB-94. Bar, 10 μm. (C) Immunofluorescent staining of collagen cross sections reveals zones of denatured collagen (stained green, arrowheads) surrounding invading fibroblasts (stained red, arrow) in the absence, but not the presence, of BB-94. Bar, 100 μm. (D) Light micrographs of SCC-1 monolayers cultured atop collagen gels (2.2 mg/ml) and stimulated with hepatocyte growth factor (HGF; 50 ng/ml) in 10% serum for 0 or 4 d in the absence or presence of BB-94. Arrows indicate islands of invasive cells. Bar, 100 μm. (E) HGF-stimulated SCC-1 cells were cultured atop three-dimensional gels of rhodamine-labeled collagen gels for 4 d in the presence of 10% serum, and tunneling behavior was assessed by laser confocal microscopy. Tunnels were not observed in the presence of BB-94. SCC-1 cells (stained with DAPI and phalloidin) invade rhodamine-labeled, pepsin-extracted collagen gels without generating tunnels. (F and G) Fibroblast (F) or tumor cell (HT1080 and SCC-1; G) invasion and collagenolytic activities were monitored in three-dimensional collagen gels (2.2 mg/ml) supplemented with 10% serum in the absence or presence of inhibitors for 6 or 4 d, respectively. 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

fig1: Collagen-invasive and -degradative activities of fibroblasts and tumor cells. (A) Transmission electron micrographs of fibroblast monolayers cultured atop three-dimensional collagen gels (2.2 mg/ml) with PDGF-BB (10 ng/ml) in the presence of 10% serum at the start of the culture period (0 d) or after 6 d in either the absence or presence of 5 μM BB-94. Zones of collagen clearing are observed in areas surrounding invading cells (middle, arrows). Double-headed arrow indicates the position of the underlying gel. Bar, 10 μm. (B) Laser confocal micrographs of rhodamine-labeled collagen gels (2.2 mg/ml) incubated with fibroblasts and PDGF in 10% serum for 0 or 6 d in the absence or presence of BB-94. Bar, 10 μm. (C) Immunofluorescent staining of collagen cross sections reveals zones of denatured collagen (stained green, arrowheads) surrounding invading fibroblasts (stained red, arrow) in the absence, but not the presence, of BB-94. Bar, 100 μm. (D) Light micrographs of SCC-1 monolayers cultured atop collagen gels (2.2 mg/ml) and stimulated with hepatocyte growth factor (HGF; 50 ng/ml) in 10% serum for 0 or 4 d in the absence or presence of BB-94. Arrows indicate islands of invasive cells. Bar, 100 μm. (E) HGF-stimulated SCC-1 cells were cultured atop three-dimensional gels of rhodamine-labeled collagen gels for 4 d in the presence of 10% serum, and tunneling behavior was assessed by laser confocal microscopy. Tunnels were not observed in the presence of BB-94. SCC-1 cells (stained with DAPI and phalloidin) invade rhodamine-labeled, pepsin-extracted collagen gels without generating tunnels. (F and G) Fibroblast (F) or tumor cell (HT1080 and SCC-1; G) invasion and collagenolytic activities were monitored in three-dimensional collagen gels (2.2 mg/ml) supplemented with 10% serum in the absence or presence of inhibitors for 6 or 4 d, respectively. Results are expressed as the mean ± 1 SEM of three or more experiments.

Mentions: After in vitro fibrillogenesis, type I collagen forms a gel-like network of interlocking fibrils stabilized by intermolecular, aldolimine cross-links similar to those generated in vivo (Hay, 1991). When cultured atop this three-dimensional substratum and exposed to a chemotactic gradient of PDGF-BB in the presence of serum, fibroblasts invade the collagen gel (Fig. 1 A). Tumor cells of mesenchymal or epithelial origin (e.g., HT-1080 and SCC-1 cells, respectively) also infiltrate the three-dimensional collagen barriers over the course of the 4-d culture period (Fig. 1 D). Coincident with the invasive process, both fibroblasts and tumor cells remodel the collagen substratum, leaving a network of tunnels that crisscross the traversed matrix (Fig. 1, B and E). Significantly, the channels formed are not the products of mechanical remodeling alone (Sawhney and Howard, 2002), as tunnel walls are lined by immuno-detectable collagen denaturation products (Fig. 1 C). Likewise, expression of the collagen-invasive phenotype by fibroblasts or tumor cells correlates with the accumulation of type I collagen degradation products (Fig. 1, F and G).


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)

Collagen-invasive and -degradative activities of fibroblasts and tumor cells. (A) Transmission electron micrographs of fibroblast monolayers cultured atop three-dimensional collagen gels (2.2 mg/ml) with PDGF-BB (10 ng/ml) in the presence of 10% serum at the start of the culture period (0 d) or after 6 d in either the absence or presence of 5 μM BB-94. Zones of collagen clearing are observed in areas surrounding invading cells (middle, arrows). Double-headed arrow indicates the position of the underlying gel. Bar, 10 μm. (B) Laser confocal micrographs of rhodamine-labeled collagen gels (2.2 mg/ml) incubated with fibroblasts and PDGF in 10% serum for 0 or 6 d in the absence or presence of BB-94. Bar, 10 μm. (C) Immunofluorescent staining of collagen cross sections reveals zones of denatured collagen (stained green, arrowheads) surrounding invading fibroblasts (stained red, arrow) in the absence, but not the presence, of BB-94. Bar, 100 μm. (D) Light micrographs of SCC-1 monolayers cultured atop collagen gels (2.2 mg/ml) and stimulated with hepatocyte growth factor (HGF; 50 ng/ml) in 10% serum for 0 or 4 d in the absence or presence of BB-94. Arrows indicate islands of invasive cells. Bar, 100 μm. (E) HGF-stimulated SCC-1 cells were cultured atop three-dimensional gels of rhodamine-labeled collagen gels for 4 d in the presence of 10% serum, and tunneling behavior was assessed by laser confocal microscopy. Tunnels were not observed in the presence of BB-94. SCC-1 cells (stained with DAPI and phalloidin) invade rhodamine-labeled, pepsin-extracted collagen gels without generating tunnels. (F and G) Fibroblast (F) or tumor cell (HT1080 and SCC-1; G) invasion and collagenolytic activities were monitored in three-dimensional collagen gels (2.2 mg/ml) supplemented with 10% serum in the absence or presence of inhibitors for 6 or 4 d, respectively. 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

fig1: Collagen-invasive and -degradative activities of fibroblasts and tumor cells. (A) Transmission electron micrographs of fibroblast monolayers cultured atop three-dimensional collagen gels (2.2 mg/ml) with PDGF-BB (10 ng/ml) in the presence of 10% serum at the start of the culture period (0 d) or after 6 d in either the absence or presence of 5 μM BB-94. Zones of collagen clearing are observed in areas surrounding invading cells (middle, arrows). Double-headed arrow indicates the position of the underlying gel. Bar, 10 μm. (B) Laser confocal micrographs of rhodamine-labeled collagen gels (2.2 mg/ml) incubated with fibroblasts and PDGF in 10% serum for 0 or 6 d in the absence or presence of BB-94. Bar, 10 μm. (C) Immunofluorescent staining of collagen cross sections reveals zones of denatured collagen (stained green, arrowheads) surrounding invading fibroblasts (stained red, arrow) in the absence, but not the presence, of BB-94. Bar, 100 μm. (D) Light micrographs of SCC-1 monolayers cultured atop collagen gels (2.2 mg/ml) and stimulated with hepatocyte growth factor (HGF; 50 ng/ml) in 10% serum for 0 or 4 d in the absence or presence of BB-94. Arrows indicate islands of invasive cells. Bar, 100 μm. (E) HGF-stimulated SCC-1 cells were cultured atop three-dimensional gels of rhodamine-labeled collagen gels for 4 d in the presence of 10% serum, and tunneling behavior was assessed by laser confocal microscopy. Tunnels were not observed in the presence of BB-94. SCC-1 cells (stained with DAPI and phalloidin) invade rhodamine-labeled, pepsin-extracted collagen gels without generating tunnels. (F and G) Fibroblast (F) or tumor cell (HT1080 and SCC-1; G) invasion and collagenolytic activities were monitored in three-dimensional collagen gels (2.2 mg/ml) supplemented with 10% serum in the absence or presence of inhibitors for 6 or 4 d, respectively. Results are expressed as the mean ± 1 SEM of three or more experiments.
Mentions: After in vitro fibrillogenesis, type I collagen forms a gel-like network of interlocking fibrils stabilized by intermolecular, aldolimine cross-links similar to those generated in vivo (Hay, 1991). When cultured atop this three-dimensional substratum and exposed to a chemotactic gradient of PDGF-BB in the presence of serum, fibroblasts invade the collagen gel (Fig. 1 A). Tumor cells of mesenchymal or epithelial origin (e.g., HT-1080 and SCC-1 cells, respectively) also infiltrate the three-dimensional collagen barriers over the course of the 4-d culture period (Fig. 1 D). Coincident with the invasive process, both fibroblasts and tumor cells remodel the collagen substratum, leaving a network of tunnels that crisscross the traversed matrix (Fig. 1, B and E). Significantly, the channels formed are not the products of mechanical remodeling alone (Sawhney and Howard, 2002), as tunnel walls are lined by immuno-detectable collagen denaturation products (Fig. 1 C). Likewise, expression of the collagen-invasive phenotype by fibroblasts or tumor cells correlates with the accumulation of type I collagen degradation products (Fig. 1, F and G).

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