The role of extracellular matrix in human astrocytoma migration and proliferation studied in a microliter scale assay

Berens, Michael E.; Rief, Monique D.; Loo, Melinda A.; Giese, Alf

doi:10.1007/bf01755884

Cited by 109 publications

(61 citation statements)

References 28 publications

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“…Radial Cell Migration Assay-Cell migration was quantified as described previously (27). Glioma cells stably expressing shRNA targeting SGEF, a control empty vector, or cells that were transfected with siRNA targeting luciferase or TRAF2 24 h post-transfection were plated onto 10-well glass slides precoated with 10 g/ml laminin.…”

Section: Methodsmentioning

confidence: 99%

The Src Homology 3 Domain-containing Guanine Nucleotide Exchange Factor Is Overexpressed in High-grade Gliomas and Promotes Tumor Necrosis Factor-like Weak Inducer of Apoptosis-Fibroblast Growth Factor-inducible 14-induced Cell Migration and Invasion via Tumor Necrosis Factor Receptor-associated Factor 2

Ensign

Mathews

Eschbacher

et al. 2013

Journal of Biological Chemistry

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Background: Glioblastoma is characterized by heightened cell invasion and therapeutic resistance. Results: The Src homology 3 domain-containing GEF (SGEF) promotes fibroblast growth factor-inducible 14 (Fn14) proinvasive signaling in glioblastoma via TNF receptor-associated factor 2 (TRAF2). Conclusion: SGEF is an important regulator of glioblastoma cell invasion downstream of Fn14. Significance: Therapy directed at mediators of invasion may confer increased chemotherapeutic and radiologic susceptibility in glioblastoma.

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Section: Methodsmentioning

confidence: 99%

The Src Homology 3 Domain-containing Guanine Nucleotide Exchange Factor Is Overexpressed in High-grade Gliomas and Promotes Tumor Necrosis Factor-like Weak Inducer of Apoptosis-Fibroblast Growth Factor-inducible 14-induced Cell Migration and Invasion via Tumor Necrosis Factor Receptor-associated Factor 2

Ensign

Mathews

Eschbacher

et al. 2013

Journal of Biological Chemistry

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“…Migration assays were done using the microliter scale radial monolayer assay as described (19,20). Briefly, 10-well Tefloncoated slides (CSM Inc., Phoenix, AZ) were coated with 10 or 0.1 Ag/mL laminin and 0.1% BSA.…”

Section: Methodsmentioning

confidence: 99%

Inhibition of Rho-Kinase Affects Astrocytoma Morphology, Motility, and Invasion through Activation of Rac1

et al. 2005

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Malignant astrocytomas are highly invasive neoplasms infiltrating diffusely into regions of normal brain. Whereas the molecular and cellular mechanisms governing astrocytoma invasion remain poorly understood, evidence in other cell systems has implicated a role for the Rho-GTPases in cell motility and invasion. Here, we examine how the inhibition or activation of Rho-kinase (ROCK) affects astrocytoma morphology, motility, and invasion. ROCK was inhibited in astrocytoma cells by using 5 to 100 Mmol/L of Y27632 or by expressing the dominant-negative ROCK mutant, RB/PH TT. ROCK activation was achieved by expressing a constitutively active mutant, CAT. ROCK inhibition led to morphologic and cytoskeletal alterations characterized by an increase in the number and length of cell processes, increased membrane ruffling, and collapse of actin stress fibers. Using twodimensional radial migration and Boyden chamber assays, we show that astrocytoma migration and invasion were increased at least 2-fold by ROCK inhibition. On the contrary, ROCK activation significantly inhibited migration and invasion of astrocytoma cells. Furthermore, using a Rac-GTP pulldown assay, we show that Rac1 is activated as a consequence of ROCK inhibition. Finally, we show that treatment of astrocytoma cells with small interfering RNA duplexes specific for Rac1-reversed stellation, prevented membrane ruffling formation and abrogated the increased motility observed following treatment with Y27632. Our data show that Rac1 plays a major role in astrocytoma morphology, motility, and invasion. These findings warrant further investigation to determine precisely how the modulation of Rac1 and ROCK can be exploited to inhibit glioma invasion.

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“…Migration Assays. Cellular migration was quantified by a microliter scale radial migration assay as described previously (14). Briefly, astrocytomaderived ECM was prepared on 10-well slides (Erie Scientific, Portsmouth, NH) as described above.…”

Section: Cell Culture Conditions and Extracellular Matrix (Ecm) Prepamentioning

confidence: 99%

The Phosphorylation of EphB2 Receptor Regulates Migration and Invasion of Human Glioma Cells

et al. 2004

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Eph receptor tyrosine kinases and their ligands, ephrins, mediate neurodevelopmental processes such as boundary formation, axon guidance, vasculogenesis, and cell migration. We determined the expression profiles of the Eph family members in five glioma cell lines under migrating and nonmigrating conditions. EphB2 mRNA was overexpressed in all five during migration (1.2-2.8-fold). We found abundant EphB2 protein as well as strong phosphorylation of EphB2 in migrating U87 cells. Confocal imaging showed EphB2 localized in lamellipodia of motile U87 cells. Treatment with ephrin-B1/Fc chimera stimulated migration and invasion of U87, whereas treatment with a blocking EphB2 antibody significantly inhibited migration and invasion. Forced expression of EphB2 in U251 cells stimulated cell migration and invasion and diminished adhesion concomitant with the tyrosine phosphorylation of EphB2. U251 stably transfected with EphB2 showed more scattered and more pronounced invasive growth in an ex vivo rat brain slice. In human brain tumor specimens, EphB2 expression was higher in glioblastomas than in lowgrade astrocytomas or normal brain; patterns of phosphorylated EphB2 matched the expression levels. Laser capture microdissection of invading glioblastoma cells revealed elevated EphB2 mRNA (1.5-3.5-fold) in 7 of 7 biopsy specimens. Immunohistochemistry demonstrated EphB2 localization primarily in glioblastoma cells (56 of 62 cases) and not in normal brain. This is the first demonstration that migrating glioblastoma cells overexpress EphB2 in vitro and in vivo; glioma migration and invasion are promoted by activation of EphB2 or inhibited by blocking EphB2. Dysregulation of EphB2 expression or function may underlie glioma invasion.

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The role of extracellular matrix in human astrocytoma migration and proliferation studied in a microliter scale assay

Cited by 109 publications

References 28 publications

Inhibition of Rho-Kinase Affects Astrocytoma Morphology, Motility, and Invasion through Activation of Rac1

The Phosphorylation of EphB2 Receptor Regulates Migration and Invasion of Human Glioma Cells

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