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Compensation mechanism in tumor cell migration: mesenchymal-amoeboid transition after blocking of pericellular proteolysis.

Wolf K, Mazo I, Leung H, Engelke K, von Andrian UH, Deryugina EI, Strongin AY, Bröcker EB, Friedl P - J. Cell Biol. (2003)

Bottom Line: This process, however, is only incompletely attenuated by protease inhibitor-based treatment, suggesting the existence of migratory compensation strategies.In three-dimensional collagen matrices, spindle-shaped proteolytically potent HT-1080 fibrosarcoma and MDA-MB-231 carcinoma cells exhibited a constitutive mesenchymal-type movement including the coclustering of beta 1 integrins and MT1-matrix metalloproteinase (MMP) at fiber bindings sites and the generation of tube-like proteolytic degradation tracks.Near-total inhibition of MMPs, serine proteases, cathepsins, and other proteases, however, induced a conversion toward spherical morphology at near undiminished migration rates.

View Article: PubMed Central - PubMed

Affiliation: Department of Dermatology, University of Würzburg, 97080 Würzburg, Germany.

ABSTRACT
Invasive tumor dissemination in vitro and in vivo involves the proteolytic degradation of ECM barriers. This process, however, is only incompletely attenuated by protease inhibitor-based treatment, suggesting the existence of migratory compensation strategies. In three-dimensional collagen matrices, spindle-shaped proteolytically potent HT-1080 fibrosarcoma and MDA-MB-231 carcinoma cells exhibited a constitutive mesenchymal-type movement including the coclustering of beta 1 integrins and MT1-matrix metalloproteinase (MMP) at fiber bindings sites and the generation of tube-like proteolytic degradation tracks. Near-total inhibition of MMPs, serine proteases, cathepsins, and other proteases, however, induced a conversion toward spherical morphology at near undiminished migration rates. Sustained protease-independent migration resulted from a flexible amoeba-like shape change, i.e., propulsive squeezing through preexisting matrix gaps and formation of constriction rings in the absence of matrix degradation, concomitant loss of clustered beta 1 integrins and MT1-MMP from fiber binding sites, and a diffuse cortical distribution of the actin cytoskeleton. Acquisition of protease-independent amoeboid dissemination was confirmed for HT-1080 cells injected into the mouse dermis monitored by intravital multiphoton microscopy. In conclusion, the transition from proteolytic mesenchymal toward nonproteolytic amoeboid movement highlights a supramolecular plasticity mechanism in cell migration and further represents a putative escape mechanism in tumor cell dissemination after abrogation of pericellular proteolysis.

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Proteolytic migration of HT-1080/MT1 fibrosarcoma cells in 3D collagen lattice. (A) Morphology of HT-1080/MT1 cell migrating through 3D collagen lattice monitored by video microscopy. (B) Reduction of migration speed (black bars) and induction of detached, nonmobile spherical morphology (inset) by adhesion-perturbing anti–β1 integrin antibody 4B4. (C) Confocal backscatter (matrix fibers) and fluorescence of MT1-MMP (red), β1 integrins (green), and colocalization (yellow; arrowheads) at fiber traction zone of the leading edge. (D) 3D reconstruction of a calcein-stained migrating cell by time-lapse confocal microscopy and (E) backscatter signal of the same cell from the central section (time, 60 min). Fiber bundling (white arrowheads), deposition of cell fragments (black arrowhead), and newly formed matrix defect (asterisk). Black arrows, direction of migration. Bars, 20 μm.
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fig1: Proteolytic migration of HT-1080/MT1 fibrosarcoma cells in 3D collagen lattice. (A) Morphology of HT-1080/MT1 cell migrating through 3D collagen lattice monitored by video microscopy. (B) Reduction of migration speed (black bars) and induction of detached, nonmobile spherical morphology (inset) by adhesion-perturbing anti–β1 integrin antibody 4B4. (C) Confocal backscatter (matrix fibers) and fluorescence of MT1-MMP (red), β1 integrins (green), and colocalization (yellow; arrowheads) at fiber traction zone of the leading edge. (D) 3D reconstruction of a calcein-stained migrating cell by time-lapse confocal microscopy and (E) backscatter signal of the same cell from the central section (time, 60 min). Fiber bundling (white arrowheads), deposition of cell fragments (black arrowhead), and newly formed matrix defect (asterisk). Black arrows, direction of migration. Bars, 20 μm.

Mentions: In 3D collagen matrices, the migration of HT-1080/MT1 cells was characterized by polarized binding of the leading edge to collagen fibers, generating traction and spindle-shaped elongation of the cell body (Fig. 1 A; Video 1 A, available online at http://www.jcb.org/cgi/content/full/jcb.200209006/DC1). Both polarization and migration were abrogated by adhesion-perturbing anti–β1 integrin mAb 4B4 (Fig. 1 B; Video 1 B), confirming β1 integrin–mediated migratory force generation. MT1-MMP and β1 integrins were coclustered at interaction sites to collagen fibers (Fig. 1 C), which represented the location of initial fiber binding, traction, and bundling toward the leading edge (Fig. 1, D and E, white arrowheads; Video 2, available online at http://www.jcb.org/cgi/content/full/jcb.200209006/DC1). Upon forward movement of the cell body, the detachment of the trailing edge generated a circumscribed matrix defect that was bordered by aligned multifibrillar collagen bundles (Fig. 1 E, asterisks; Video 2). Cell detachment was further accompanied by the shedding of surface determinants and cell fragments along the migration track (Fig. 1 D, black arrowhead). This mesenchymal migration type of spindle-shaped cells that produce matrix defects is consistent with the integrin-dependent haptokinetic migration reported for fibroblasts (Doane and Birk, 1991; Palecek et al., 1997) and further supports the concept of focalized pericellular proteolysis for the generation of migration tracks (Murphy and Gavrilovic, 1999).


Compensation mechanism in tumor cell migration: mesenchymal-amoeboid transition after blocking of pericellular proteolysis.

Wolf K, Mazo I, Leung H, Engelke K, von Andrian UH, Deryugina EI, Strongin AY, Bröcker EB, Friedl P - J. Cell Biol. (2003)

Proteolytic migration of HT-1080/MT1 fibrosarcoma cells in 3D collagen lattice. (A) Morphology of HT-1080/MT1 cell migrating through 3D collagen lattice monitored by video microscopy. (B) Reduction of migration speed (black bars) and induction of detached, nonmobile spherical morphology (inset) by adhesion-perturbing anti–β1 integrin antibody 4B4. (C) Confocal backscatter (matrix fibers) and fluorescence of MT1-MMP (red), β1 integrins (green), and colocalization (yellow; arrowheads) at fiber traction zone of the leading edge. (D) 3D reconstruction of a calcein-stained migrating cell by time-lapse confocal microscopy and (E) backscatter signal of the same cell from the central section (time, 60 min). Fiber bundling (white arrowheads), deposition of cell fragments (black arrowhead), and newly formed matrix defect (asterisk). Black arrows, direction of migration. Bars, 20 μm.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2172637&req=5

fig1: Proteolytic migration of HT-1080/MT1 fibrosarcoma cells in 3D collagen lattice. (A) Morphology of HT-1080/MT1 cell migrating through 3D collagen lattice monitored by video microscopy. (B) Reduction of migration speed (black bars) and induction of detached, nonmobile spherical morphology (inset) by adhesion-perturbing anti–β1 integrin antibody 4B4. (C) Confocal backscatter (matrix fibers) and fluorescence of MT1-MMP (red), β1 integrins (green), and colocalization (yellow; arrowheads) at fiber traction zone of the leading edge. (D) 3D reconstruction of a calcein-stained migrating cell by time-lapse confocal microscopy and (E) backscatter signal of the same cell from the central section (time, 60 min). Fiber bundling (white arrowheads), deposition of cell fragments (black arrowhead), and newly formed matrix defect (asterisk). Black arrows, direction of migration. Bars, 20 μm.
Mentions: In 3D collagen matrices, the migration of HT-1080/MT1 cells was characterized by polarized binding of the leading edge to collagen fibers, generating traction and spindle-shaped elongation of the cell body (Fig. 1 A; Video 1 A, available online at http://www.jcb.org/cgi/content/full/jcb.200209006/DC1). Both polarization and migration were abrogated by adhesion-perturbing anti–β1 integrin mAb 4B4 (Fig. 1 B; Video 1 B), confirming β1 integrin–mediated migratory force generation. MT1-MMP and β1 integrins were coclustered at interaction sites to collagen fibers (Fig. 1 C), which represented the location of initial fiber binding, traction, and bundling toward the leading edge (Fig. 1, D and E, white arrowheads; Video 2, available online at http://www.jcb.org/cgi/content/full/jcb.200209006/DC1). Upon forward movement of the cell body, the detachment of the trailing edge generated a circumscribed matrix defect that was bordered by aligned multifibrillar collagen bundles (Fig. 1 E, asterisks; Video 2). Cell detachment was further accompanied by the shedding of surface determinants and cell fragments along the migration track (Fig. 1 D, black arrowhead). This mesenchymal migration type of spindle-shaped cells that produce matrix defects is consistent with the integrin-dependent haptokinetic migration reported for fibroblasts (Doane and Birk, 1991; Palecek et al., 1997) and further supports the concept of focalized pericellular proteolysis for the generation of migration tracks (Murphy and Gavrilovic, 1999).

Bottom Line: This process, however, is only incompletely attenuated by protease inhibitor-based treatment, suggesting the existence of migratory compensation strategies.In three-dimensional collagen matrices, spindle-shaped proteolytically potent HT-1080 fibrosarcoma and MDA-MB-231 carcinoma cells exhibited a constitutive mesenchymal-type movement including the coclustering of beta 1 integrins and MT1-matrix metalloproteinase (MMP) at fiber bindings sites and the generation of tube-like proteolytic degradation tracks.Near-total inhibition of MMPs, serine proteases, cathepsins, and other proteases, however, induced a conversion toward spherical morphology at near undiminished migration rates.

View Article: PubMed Central - PubMed

Affiliation: Department of Dermatology, University of Würzburg, 97080 Würzburg, Germany.

ABSTRACT
Invasive tumor dissemination in vitro and in vivo involves the proteolytic degradation of ECM barriers. This process, however, is only incompletely attenuated by protease inhibitor-based treatment, suggesting the existence of migratory compensation strategies. In three-dimensional collagen matrices, spindle-shaped proteolytically potent HT-1080 fibrosarcoma and MDA-MB-231 carcinoma cells exhibited a constitutive mesenchymal-type movement including the coclustering of beta 1 integrins and MT1-matrix metalloproteinase (MMP) at fiber bindings sites and the generation of tube-like proteolytic degradation tracks. Near-total inhibition of MMPs, serine proteases, cathepsins, and other proteases, however, induced a conversion toward spherical morphology at near undiminished migration rates. Sustained protease-independent migration resulted from a flexible amoeba-like shape change, i.e., propulsive squeezing through preexisting matrix gaps and formation of constriction rings in the absence of matrix degradation, concomitant loss of clustered beta 1 integrins and MT1-MMP from fiber binding sites, and a diffuse cortical distribution of the actin cytoskeleton. Acquisition of protease-independent amoeboid dissemination was confirmed for HT-1080 cells injected into the mouse dermis monitored by intravital multiphoton microscopy. In conclusion, the transition from proteolytic mesenchymal toward nonproteolytic amoeboid movement highlights a supramolecular plasticity mechanism in cell migration and further represents a putative escape mechanism in tumor cell dissemination after abrogation of pericellular proteolysis.

Show MeSH
Related in: MedlinePlus