Human malignant glioblastomas are highly invasive tumors. Increased cell motility and degradation of the surrounding extracellular matrix are essential for tumor invasion. PI3K/Akt signaling pathway emerges as a common pathway regulating cellular proliferation, migration and invasion; however, its contribution to particular process and downstream cascades remain poorly defined. We have previously demonstrated that Cyclosporin A (CsA) affects glioblastoma invasion in organotypic brain slices and tumorigenicity in mice. Here we show that CsA impairs migration and invasion of human glioblastoma cells by downregulation of Akt phosphorylation. Interference with PI-3K/Akt signaling was crucial for CsA effect on invasion, because overexpression of constitutively active myr-Akt antagonized drug action. Furthermore, the drug was not effective in T98G glioblastoma cells with constitutively high level of phosphorylated Akt. CsA, comparably to pharmacological inhibitors of PI3K/Akt signaling (LY294002, A443654), reduced motility of glioblastoma cells, diminished MMP-2 gelatinolytic activity and MMP-2 and MT1-MMP expression. The latter effect was mimicked by overexpression of dominant negative Akt mutants. We demonstrate that CsA and LY294002 reduced MMP transcription partly via modulation of IkappaB phosphorylation and NFkappaB transcriptional activity. Those effects were not mediated by inhibition of calcineurin, a classical CsA target. Additionally, CsA reduced phosphorylation and activity of focal adhesion kinase that was associated with rapid morphological alterations, rearrangement of lamellipodia and impairment of MT1-MMP translocation to membrane protrusions. Our results document novel, Akt-dependent mechanisms of interference with motility/invasion of human glioblastoma cells: through a rapid modulation of cell adhesion and MT1-MMP translocation to membrane protrusions and delayed, partly NFkappaB-dependent, downregulation of MMP-2 and MT1-MMP expression. This article is part of a Special Issue entitled: 11th European Symposium on Calcium.
Downregulation of Akt and FAK phosphorylation reduces invasion of glioblastoma cells by impairment of MT1-MMP shuttling to lamellipodia and downregulates MMPs expression
Kwiatkowska, A.; Kijewska, M.; Lipko, M.; Hibner, U.; Kaminska, B.
Biochim Biophys Acta
2011-05 / vol 1813 / pages 655-67
10.1016/j.bbamcr.2011.01.020 S0167-4889(11)00028-0 [pii]
0006-3002 (Print) 0006-3002 (Linking)
Cell Line, Tumor; Humans; Animals; Mice; Cell Shape/drug effects; *Down-Regulation/drug effects; Actins/metabolism; Cell Movement/drug effects; Cyclosporine/pharmacology; Focal Adhesion Kinase 1/*metabolism; Glioblastoma/*enzymology/pathology; Matrix Metalloproteinase 14/*metabolism; Matrix Metalloproteinase 2/*metabolism; Neoplasm Invasiveness; NF-kappa B/genetics; Phosphatidylinositol 3-Kinases/antagonists & inhibitors/metabolism; Phosphorylation/drug effects; Protein Transport/drug effects; Proto-Oncogene Proteins c-akt/antagonists & inhibitors/*metabolism; Pseudopodia/drug effects/*enzymology; Signal Transduction/drug effects; Transcription, Genetic/drug effects