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Membrane-type 1 matrix metalloprotease (MT1-MMP) enables invasive migration of glioma cells in central nervous system white matter.

Beliën AT, Paganetti PA, Schwab ME - J. Cell Biol. (1999)

Bottom Line: Plasma membranes of both MT1-MMP-transfected fibroblasts and C6 glioma cells inactivated inhibitory myelin extracts, and this activity was sensitive to the same protease inhibitors.Interestingly, pretreatment of CNS myelin with gelatinase A/MMP-2 could not inactivate its inhibitory property.These data imply an important role of MT1-MMP in spreading and migration of glioma cells on white matter constituents in vitro and point to a function of MT1-MMP in the invasive behavior of malignant gliomas in the CNS in vivo.

View Article: PubMed Central - PubMed

Affiliation: Brain Research Institute, University of Zurich and Swiss Federal Institute of Technology, 8057 Zurich, Switzerland. belien@hifo.unizh.ch

ABSTRACT
Invasive glioma cells migrate preferentially along central nervous system (CNS) white matter fiber tracts irrespective of the fact that CNS myelin contains proteins that inhibit cell migration and neurite outgrowth. Previous work has demonstrated that to migrate on a myelin substrate and to overcome its inhibitory effect, rat C6 and human glioblastoma cells require a membrane-bound metalloproteolytic activity (C6-MP) which shares several biochemical and pharmacological characteristics with MT1-MMP. We show now that MT1-MMP is expressed on the surface of rat C6 glioblastoma cells and is coenriched with C6-MP activity. Immunodepletion of C6-MP activity is achieved with an anti-MT1-MMP antibody. These data suggest that MT1-MMP and the C6-MP are closely related or identical. When mouse 3T3 fibroblasts were transfected with MT1-MMP they acquired the ability to spread and migrate on the nonpermissive myelin substrate and to infiltrate into adult rat optic nerve explants. MT1-MMP-transfected fibroblasts and C6 glioma cells were able to digest bNI-220, one of the most potent CNS myelin inhibitory proteins. Plasma membranes of both MT1-MMP-transfected fibroblasts and C6 glioma cells inactivated inhibitory myelin extracts, and this activity was sensitive to the same protease inhibitors. Interestingly, pretreatment of CNS myelin with gelatinase A/MMP-2 could not inactivate its inhibitory property. These data imply an important role of MT1-MMP in spreading and migration of glioma cells on white matter constituents in vitro and point to a function of MT1-MMP in the invasive behavior of malignant gliomas in the CNS in vivo.

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Effect of various metalloprotease inhibitors on the  spreading ability of MT1-MMP–transfected fibroblasts and C6  glioma cells on CNS myelin. (a) Transiently MT1-MMP–transfected fibroblasts of which only ∼15% express high MT1-MMP  levels. The proportion of spreading cells on CNS myelin substrate  was reduced by 15% by 1 mM EDTA, 200 μM o-phenanthroline  (phen), 300 μM phosphoramidon (phos), 10 μM Cbz-Phe-Ala-Phe-Tyr-amide (FAFY), and 100 nM TIMP2. No inhibition was  observed in presence of 1 μM BB94 or 100 nM TIMP1. (b)  About 80% of the C6 cells spread on the inhibitory CNS myelin  substrate. This was reduced to ∼25% by the same inhibitors  found to affect the MT1-MMP–transfected fibroblasts. Values  are means ± SEM of three experiments, performed in triplicate.  Significance with the two-tailed Student's t test: P < 0.01 (**).
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Figure 6: Effect of various metalloprotease inhibitors on the spreading ability of MT1-MMP–transfected fibroblasts and C6 glioma cells on CNS myelin. (a) Transiently MT1-MMP–transfected fibroblasts of which only ∼15% express high MT1-MMP levels. The proportion of spreading cells on CNS myelin substrate was reduced by 15% by 1 mM EDTA, 200 μM o-phenanthroline (phen), 300 μM phosphoramidon (phos), 10 μM Cbz-Phe-Ala-Phe-Tyr-amide (FAFY), and 100 nM TIMP2. No inhibition was observed in presence of 1 μM BB94 or 100 nM TIMP1. (b) About 80% of the C6 cells spread on the inhibitory CNS myelin substrate. This was reduced to ∼25% by the same inhibitors found to affect the MT1-MMP–transfected fibroblasts. Values are means ± SEM of three experiments, performed in triplicate. Significance with the two-tailed Student's t test: P < 0.01 (**).

Mentions: The effects of several metalloprotease inhibitors on the spreadings of MT1-MMP–transfected fibroblasts (Fig. 6 a) and of C6 glioblastoma cells (Fig. 6 b) plated on an inhibitory CNS protein substrate were studied. In contrast to normal 3T3 fibroblasts (Fig. 2 b, Fig. 4, and Fig. 5 b), MT1-MMP–transfected fibroblasts as well as C6 cells show a large proportion of spreading cells. Addition of the chelators EDTA and o-phenanthroline greatly reduced the spreading ability of both cell types (P < 0.01). The more specific C6-MP inhibitors phosphoramidon and cbz-FAFY-amide (Amberger et al., 1994) also significantly reduced the spreading ability of MT1-MMP–transfected fibroblasts similar to what was observed with the C6 glioblastoma cells.


Membrane-type 1 matrix metalloprotease (MT1-MMP) enables invasive migration of glioma cells in central nervous system white matter.

Beliën AT, Paganetti PA, Schwab ME - J. Cell Biol. (1999)

Effect of various metalloprotease inhibitors on the  spreading ability of MT1-MMP–transfected fibroblasts and C6  glioma cells on CNS myelin. (a) Transiently MT1-MMP–transfected fibroblasts of which only ∼15% express high MT1-MMP  levels. The proportion of spreading cells on CNS myelin substrate  was reduced by 15% by 1 mM EDTA, 200 μM o-phenanthroline  (phen), 300 μM phosphoramidon (phos), 10 μM Cbz-Phe-Ala-Phe-Tyr-amide (FAFY), and 100 nM TIMP2. No inhibition was  observed in presence of 1 μM BB94 or 100 nM TIMP1. (b)  About 80% of the C6 cells spread on the inhibitory CNS myelin  substrate. This was reduced to ∼25% by the same inhibitors  found to affect the MT1-MMP–transfected fibroblasts. Values  are means ± SEM of three experiments, performed in triplicate.  Significance with the two-tailed Student's t test: P < 0.01 (**).
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Figure 6: Effect of various metalloprotease inhibitors on the spreading ability of MT1-MMP–transfected fibroblasts and C6 glioma cells on CNS myelin. (a) Transiently MT1-MMP–transfected fibroblasts of which only ∼15% express high MT1-MMP levels. The proportion of spreading cells on CNS myelin substrate was reduced by 15% by 1 mM EDTA, 200 μM o-phenanthroline (phen), 300 μM phosphoramidon (phos), 10 μM Cbz-Phe-Ala-Phe-Tyr-amide (FAFY), and 100 nM TIMP2. No inhibition was observed in presence of 1 μM BB94 or 100 nM TIMP1. (b) About 80% of the C6 cells spread on the inhibitory CNS myelin substrate. This was reduced to ∼25% by the same inhibitors found to affect the MT1-MMP–transfected fibroblasts. Values are means ± SEM of three experiments, performed in triplicate. Significance with the two-tailed Student's t test: P < 0.01 (**).
Mentions: The effects of several metalloprotease inhibitors on the spreadings of MT1-MMP–transfected fibroblasts (Fig. 6 a) and of C6 glioblastoma cells (Fig. 6 b) plated on an inhibitory CNS protein substrate were studied. In contrast to normal 3T3 fibroblasts (Fig. 2 b, Fig. 4, and Fig. 5 b), MT1-MMP–transfected fibroblasts as well as C6 cells show a large proportion of spreading cells. Addition of the chelators EDTA and o-phenanthroline greatly reduced the spreading ability of both cell types (P < 0.01). The more specific C6-MP inhibitors phosphoramidon and cbz-FAFY-amide (Amberger et al., 1994) also significantly reduced the spreading ability of MT1-MMP–transfected fibroblasts similar to what was observed with the C6 glioblastoma cells.

Bottom Line: Plasma membranes of both MT1-MMP-transfected fibroblasts and C6 glioma cells inactivated inhibitory myelin extracts, and this activity was sensitive to the same protease inhibitors.Interestingly, pretreatment of CNS myelin with gelatinase A/MMP-2 could not inactivate its inhibitory property.These data imply an important role of MT1-MMP in spreading and migration of glioma cells on white matter constituents in vitro and point to a function of MT1-MMP in the invasive behavior of malignant gliomas in the CNS in vivo.

View Article: PubMed Central - PubMed

Affiliation: Brain Research Institute, University of Zurich and Swiss Federal Institute of Technology, 8057 Zurich, Switzerland. belien@hifo.unizh.ch

ABSTRACT
Invasive glioma cells migrate preferentially along central nervous system (CNS) white matter fiber tracts irrespective of the fact that CNS myelin contains proteins that inhibit cell migration and neurite outgrowth. Previous work has demonstrated that to migrate on a myelin substrate and to overcome its inhibitory effect, rat C6 and human glioblastoma cells require a membrane-bound metalloproteolytic activity (C6-MP) which shares several biochemical and pharmacological characteristics with MT1-MMP. We show now that MT1-MMP is expressed on the surface of rat C6 glioblastoma cells and is coenriched with C6-MP activity. Immunodepletion of C6-MP activity is achieved with an anti-MT1-MMP antibody. These data suggest that MT1-MMP and the C6-MP are closely related or identical. When mouse 3T3 fibroblasts were transfected with MT1-MMP they acquired the ability to spread and migrate on the nonpermissive myelin substrate and to infiltrate into adult rat optic nerve explants. MT1-MMP-transfected fibroblasts and C6 glioma cells were able to digest bNI-220, one of the most potent CNS myelin inhibitory proteins. Plasma membranes of both MT1-MMP-transfected fibroblasts and C6 glioma cells inactivated inhibitory myelin extracts, and this activity was sensitive to the same protease inhibitors. Interestingly, pretreatment of CNS myelin with gelatinase A/MMP-2 could not inactivate its inhibitory property. These data imply an important role of MT1-MMP in spreading and migration of glioma cells on white matter constituents in vitro and point to a function of MT1-MMP in the invasive behavior of malignant gliomas in the CNS in vivo.

Show MeSH
Related in: MedlinePlus