Glioma invasion in vitro: regulation by matrix metalloprotease-2 and protein kinase C.
A hallmark of invasive tumors is their ability to effect degradation of the surrounding extracellular matrix (ECM) by the local production of proteolytic enzymes, such as the matrix metalloproteases (MMPs). In this paper, we demonstrate that the invasion of human gliomas is mediated by a 72 kDa MMP, referred to as MMP-2, and provide further evidence that the activity of MMP-2 is regulated by protein kinase C (PKC). The invasiveness of five human glioma cell lines (A172, U87, U118, U251, U563) was assessed in an in vitro invasion assay and was found to correlate with the level of MMP-2 activity (r2 = 0.95); in contrast, the activity of this 72 kDa metalloprotease was barely detectable in non-invasive control glial cells (non-transformed human astrocytes and oligodendrocytes). Treatment with 1,10-phenanthroline, a metalloprotease inhibitor, or with a synthetic dipeptide, containing a blocking sequence (ala-phe) specific for MMPs, resulted in a > 90% reduction in glioma invasion. Furthermore, this MMP-2 activity could be inhibited by the treatment of tumor cells with calphostin C, a specific inhibitor of PKC. Glioma cell lines treated with calphostin C demonstrated a dose-dependent decrease (IC50 = 30 nM) in tumor invasiveness with a concomitant reduction in the activity of the MMP-2. Conversely, treatment of non-invasive control astrocytes with a PKC activator (phorbol ester) led to a corresponding increase in their invasiveness and metalloprotease activity. These findings support the postulate that MMP-2 activity constitutes an important effector of human glioma invasion and that the regulation of this proteolytic activity can be modulated by PKC.
Uhm
J H
JH
Brain Tumor Research Group, Montreal Neurological Institute, Quebec, Canada.
Dooley
N P
NP
Villemure
J G
JG
Yong
V W
VW
D002455:Cell Division; D003094:Collagen / Q000378:metabolism; D017364:Collagenases / Q000378:metabolism; D004338:Drug Combinations; D004791:Enzyme Inhibitors / Q000494:pharmacology; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D018093:Gelatinases / Q000037:antagonists & inhibitors / Q000502:physiology*; D005910:Glioma / Q000201:enzymology / Q000473:pathology*; D006801:Humans; D007797:Laminin / Q000378:metabolism; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors / Q000502:physiology*; D009281:Naphthalenes / Q000494:pharmacology; D009361:Neoplasm Invasiveness; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000502:physiology*; D011509:Proteoglycans / Q000378:metabolism; D014407:Tumor Cells, Cultured
D004338:Drug Combinations; D004791:Enzyme Inhibitors; D016326:Extracellular Matrix Proteins; D007797:Laminin; D009281:Naphthalenes; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen; D011493:Protein Kinase C; D017364:Collagenases; D018093:Gelatinases; D008666:Metalloendopeptidases; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; C058819:calphostin C