Artikel
The EphB4 receptor Tyrosine kinase mediates glioma cell migration and invasion
Die Rezeptor-Tyrosine-Kinase EphB4 vermittelt Migration und Invasion von Gliom-Zellen
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Autoren
Veröffentlicht: | 8. Mai 2006 |
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Gliederung
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Objective: The Eph receptors have been primarily discovered as proteins involved in development of the nervous and vascular system, mediating cell migration and cell guidance. The aim of the present study was to investigate the role of EphB4 in glioma cell migration and invasion in vitro.
Methods: Four clones of the SF126 glioma cell line were established: wt (coding for full length EphB4 receptor), dn (signaling defective receptor), siRNA (reduced expression of endogenous EphB4) and mocks (empty vector). EphB4 wt and dn expression were characterized by RT-PCR, immunoblotting and immunocytochemistry. Phosphoblots were performed to determine EphB4 activation levels. Cell migration was assessed by the spheroid migration assay which was performed on plastic as well as on different extracellular matrix coatings (collagen IV, fibronectin, tenascin-C).
Results: Cell clones demonstrated adequate expression of EphB4 wt and dn as well as inhibition of EphB4 signaling in the dn and siRNA group. Immunocytochemistry demonstrated EphB4wt and Eph4dn expression on the cell surface of tumor cells. The migration assays performed on collagen IV, tenascin-C, and fibronectin showed increased migration of wt and dn cells when compared to mock (up to x1.5, x1.8, x3.2 fold increase, respectively, all p<0.05).
Conclusions: The results of this study indicate that the extracellular domain of EphB4 may contribute to the regulation of glioma cell migration. An interplay with ECM molecules may modulate its impact on cell migration.