Protein kinase C-mediated in vitro invasion of human glioma cells through extracellular-signal-regulated kinase and ornithine decarboxylase
| Autor: | A. B. da Rocha, C. de Lima, Vitor Monteiro, Jose Claudio Fonseca Moreira, Richard Rodnight, Andreia Kist Fernandes, Guido Lenz, Danielle Goncalves, Dennis Ricardo August Mans, Algemir Lunardi Brunetto, Gilberto Schwartsmann |
|---|---|
| Rok vydání: | 2001 |
| Předmět: |
Eflornithine
Indoles Matrix Metalloproteinases Membrane-Associated Mitogen-activated protein kinase kinase Naphthalenes Ornithine Decarboxylase Pathology and Forensic Medicine MAP2K7 Maleimides chemistry.chemical_compound Tumor Cells Cultured Humans Calphostin Neoplasm Invasiveness RNA Messenger Molecular Biology Protein kinase C Protein Kinase C Flavonoids biology Chemistry Cyclin-dependent kinase 4 Cyclin-dependent kinase 2 Metalloendopeptidases Cell Biology General Medicine Glioma Molecular biology Cell biology Calphostin C Culture Media Conditioned biology.protein Matrix Metalloproteinase 2 Tetradecanoylphorbol Acetate Cyclin-dependent kinase 9 Mitogen-Activated Protein Kinases |
| Zdroj: | Pathobiology : journal of immunopathology, molecular and cellular biology. 68(3) |
| ISSN: | 1015-2008 |
| Popis: | We investigated the involvement of protein kinase C (PKC) in the in vitro invasiveness of the A-172, U-87 and U-373 human glioma cell lines, as well as the role of ornithine decarboxylase (ODC) and/or extracellular-signal-regulated kinase (ERK) in the actions of PKC. Thus, cells were treated under serum-free conditions with the PKC activator phorbol 12-myristate 13-acetate (PMA), or with the PKC inhibitors bisindolylmaleimide I (GF 109203X) or calphostin C in the absence or presence of the ODC inhibitor D,L-α-difluoromethylornithine (DFMO), and/or the mitogen-activated protein kinase/extracellular-signal-regulated kinase inhibitor 2′-amino-3′-methoxyflavone (PD 098059). Subsequently, cells were assessed for membrane-type 1 matrix metalloproteinase (MT1-MMP) mRNA contents, 72-kD latent, and 59/62-kD activated matrix metalloproteinase 2 (MMP-2) in conditioned media, as well as invasiveness. For these purposes, we used Northern blot analysis, gelatine zymography, and an in vitro filter invasion assay, respectively. Data were related to those found with untreated cells. PKC activity was 2- to 3-fold stimulated by PMA (100 nM for 30 min), and about 2-fold inhibited by calphostin C (40 nM for 2 h) or GF 109203X (5 µM for 20 min). This was accompanied by a similar increase or decrease, respectively, in MT1-MMP mRNA expression, 59/62-kD MMP-2 activity, and in vitro invasion. Inhibition of ODC activity (about 2-fold by 24 h DFMO 5 mM), ERK activation (almost completely by 20 min PD 098059 50 µM), or both these enzymes simultaneously led to a reduction by about half in levels of MT1-MMP mRNA, 59/62-kD MMP-2 activity, and invasion in untreated as well as PMA-stimulated cells. The use of these compounds did not significantly alter the inhibitory effects of GF 109203X or calphostin C. Modulation of PKC and/or ERK activity resulted in corresponding changes in ERK and/or ODC activities, but interference with ODC affected neither ERK nor PKC. Our data suggest a regulatory role for PKC, in co-operation with ERK and ODC, in glioma cell invasion, by modulation of MT1-MMP mRNA expression and MMP-2 activation. |
| Databáze: | OpenAIRE |
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