CD44 expression and MMP-2 secretion by mouse glioma cells: effect of interferon and anti-CD44 antibody.
We have previously reported that invasiveness of mouse glioma G-26, which expresses CD44 adhesion molecule, was inhibited in vitro following treatment with anti-CD44 antibody or mouse interferon alpha/beta (MuIFN alpha/beta). Here, we evaluated whether the expression of transmembrane CD44 adhesion molecule and/or secretion of extracellular matrix metalloproteinases (MMPs) were affected when glioma cell invasion was inhibited. Flow cytometric evaluation of CD44 adhesion molecule expression in G-26 glioma using anti-CD44 antibody, confirmed that G-26 cells were CD44+. Following 3-day treatment with MuIFN alpha/beta at 8 x 10(2) or 8 x 10(3) IU/ml of glioma cells, the expression of CD44 was not significantly affected as reflected by CD44+ cell number and fluorescence intensity. The pretreatment of glioma cells for 1 day with anti-CD44 antibody resulted in a 30-60% decrease of CD44 expression. This coincided with significantly (p < 0.05) lower cell activity as judged by MTT assay for mitochondrial activity. The zymographic evaluation of MMP activity in the G-26 glioma cell culture showed a high level of the active form of MMP-2. This level of MMP-2 was decreased following 3 day treatment of G-26 glioma cells with either 8 x 10(2) or 8 x 10(3) IU/ml of MuIFN alpha/beta but only the latter concentration produced statistically significant 55% decrease. However, following a 1 day treatment of G-26 glioma cells with anti-CD44 antibody, the level of active MMP-2 form was not significantly affected. These findings indicate that while the inhibitory effect of IFN on glioma invasion was accompanied by a decreased level of the active form of MMP-2 released extracellularly, the expression of the transmembrane CD44 adhesion molecule was not affected. Conversely, anti-CD44 antibody pretreatment of G-26 glioma, which led to the inhibition of glioma invasion, resulted in decreased CD44 expression and lower cell activity but had no effect on the MMP-2.
Wiranowska
M
M
Department of Anatomy, H. Lee Moffitt Cancer Center and Research Institute, Neuro-Oncology Program and the Neuroscience Program, University of South Florida, College of Medicine, 12901 Bruce B. Downs Blvd., Tampa, Fl. 33612, USA.
Rojiani
A M
AM
Gottschall
P E
PE
Moscinski
L C
LC
Johnson
J
J
Saporta
S
S
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000494:pharmacology*; D005434:Flow Cytometry; D005910:Glioma / Q000378:metabolism* / Q000473:pathology; D018960:Hyaluronan Receptors / Q000276:immunology / Q000378:metabolism*; D016898:Interferon-alpha / Q000494:pharmacology*; D016899:Interferon-beta / Q000494:pharmacology*; D020778:Matrix Metalloproteinase 2 / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D009361:Neoplasm Invasiveness; D009363:Neoplasm Proteins / Q000276:immunology / Q000378:metabolism*
D000888:Antibodies, Anti-Idiotypic; D018960:Hyaluronan Receptors; D016898:Interferon-alpha; D009363:Neoplasm Proteins; D016899:Interferon-beta; D020778:Matrix Metalloproteinase 2