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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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(a) Spreading of MT1-MMP–transfected fibroblasts on  culture dishes coated with inhibitory CNS membrane protein extracts. 1 h after plating ∼35% of the MT1-MMP–transfected cells  were well attached and showed a flat appearance (arrow). The  nonspreading cells appear as round, phase-bright cells. (b) Quantification of the spreading of MT1-MMP–, mock-, and nontransfected fibroblast on CNS inhibitory substrate and control  substrate (PLYS). Significantly more MT1-MMP–transfected fibroblasts spread on CNS myelin as compared with mock- or nontransfected fibroblasts. In all three conditions, cells spread  equally well on the control substrate PLYS. 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 5: (a) Spreading of MT1-MMP–transfected fibroblasts on culture dishes coated with inhibitory CNS membrane protein extracts. 1 h after plating ∼35% of the MT1-MMP–transfected cells were well attached and showed a flat appearance (arrow). The nonspreading cells appear as round, phase-bright cells. (b) Quantification of the spreading of MT1-MMP–, mock-, and nontransfected fibroblast on CNS inhibitory substrate and control substrate (PLYS). Significantly more MT1-MMP–transfected fibroblasts spread on CNS myelin as compared with mock- or nontransfected fibroblasts. In all three conditions, cells spread equally well on the control substrate PLYS. Values are means ± SEM of three experiments, performed in triplicate. Significance with the two-tailed Student's t test: P < 0.01 (**).

Mentions: To evaluate the ability of MT1-MMP–transfected, mock-transfected, and nontransfected fibroblasts to spread on an inhibitory CNS membrane protein extract (Spillmann et al., 1998), the cells were plated on this substrate, fixed after 1 h of incubation, and then the percentage of flat, well-attached, spread cells was determined (Fig. 5). Of the nontransfected and mock-transfected fibroblasts 19 ± 2 and 20 ± 2%, respectively, of the cells were able to spread on the CNS protein substrate. Upon transfection with MT1-MMP the percentage of spread cells increased to 35 ± 3% (Fig. 5 b). This increase of 15% correlates well with the transfection rate of about 15% as seen in immunofluorescence for MT1-MMP. On the control substrate (PLYS), MT1-MMP–, mock-, and nontransfected fibroblasts spread equally well.


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)

(a) Spreading of MT1-MMP–transfected fibroblasts on  culture dishes coated with inhibitory CNS membrane protein extracts. 1 h after plating ∼35% of the MT1-MMP–transfected cells  were well attached and showed a flat appearance (arrow). The  nonspreading cells appear as round, phase-bright cells. (b) Quantification of the spreading of MT1-MMP–, mock-, and nontransfected fibroblast on CNS inhibitory substrate and control  substrate (PLYS). Significantly more MT1-MMP–transfected fibroblasts spread on CNS myelin as compared with mock- or nontransfected fibroblasts. In all three conditions, cells spread  equally well on the control substrate PLYS. 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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Related In: Results  -  Collection

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Figure 5: (a) Spreading of MT1-MMP–transfected fibroblasts on culture dishes coated with inhibitory CNS membrane protein extracts. 1 h after plating ∼35% of the MT1-MMP–transfected cells were well attached and showed a flat appearance (arrow). The nonspreading cells appear as round, phase-bright cells. (b) Quantification of the spreading of MT1-MMP–, mock-, and nontransfected fibroblast on CNS inhibitory substrate and control substrate (PLYS). Significantly more MT1-MMP–transfected fibroblasts spread on CNS myelin as compared with mock- or nontransfected fibroblasts. In all three conditions, cells spread equally well on the control substrate PLYS. Values are means ± SEM of three experiments, performed in triplicate. Significance with the two-tailed Student's t test: P < 0.01 (**).
Mentions: To evaluate the ability of MT1-MMP–transfected, mock-transfected, and nontransfected fibroblasts to spread on an inhibitory CNS membrane protein extract (Spillmann et al., 1998), the cells were plated on this substrate, fixed after 1 h of incubation, and then the percentage of flat, well-attached, spread cells was determined (Fig. 5). Of the nontransfected and mock-transfected fibroblasts 19 ± 2 and 20 ± 2%, respectively, of the cells were able to spread on the CNS protein substrate. Upon transfection with MT1-MMP the percentage of spread cells increased to 35 ± 3% (Fig. 5 b). This increase of 15% correlates well with the transfection rate of about 15% as seen in immunofluorescence for MT1-MMP. On the control substrate (PLYS), MT1-MMP–, mock-, and nontransfected fibroblasts spread equally well.

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