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CD44 expression and MMP-2 secretion by mouse glioma cells: effect of interferon and anti-CD44 antibody
M Wiranowska1, A M Rojiani, P E Gottschall
1Department 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.
Abstract:
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.
Insights
Mouse interferon (MuIFN alpha/beta) inhibited glioma invasion by reducing matrix metalloproteinase-2 (MMP-2) without affecting CD44 expression. Anti-CD44 antibody also inhibited invasion, decreasing CD44 expression and cell activity but not MMP-2 levels.
Area of Science:
- Neuroscience
- Immunology
- Biochemistry
Background:
- Glioma cell invasion is a critical factor in tumor progression and metastasis.
- The CD44 adhesion molecule and matrix metalloproteinases (MMPs) are implicated in cancer cell invasion.
- Previous studies showed that mouse interferon alpha/beta (MuIFN alpha/beta) and anti-CD44 antibody inhibit G-26 mouse glioma cell invasiveness.
Purpose of the Study:
- To investigate the effects of MuIFN alpha/beta and anti-CD44 antibody on CD44 expression and MMP secretion in G-26 glioma cells.
- To elucidate the molecular mechanisms underlying the inhibition of glioma cell invasion by these agents.
Main Methods:
- Flow cytometry was used to assess CD44 expression on G-26 glioma cells.
- Zymography was employed to evaluate the activity of MMP-2 in cell culture supernatants.
- MTT assay was performed to measure cell activity.
Main Results:
- MuIFN alpha/beta treatment (3 days) significantly decreased active MMP-2 levels in a dose-dependent manner but did not affect CD44 expression.
- Anti-CD44 antibody treatment (1 day) reduced CD44 expression and cell activity but had no significant impact on active MMP-2 levels.
- Both treatments inhibited G-26 glioma cell invasion in vitro.
Conclusions:
- The inhibitory effect of MuIFN alpha/beta on glioma invasion is associated with reduced extracellular MMP-2 activity, independent of CD44 expression changes.
- Anti-CD44 antibody inhibits glioma invasion by downregulating CD44 expression and reducing cell activity, without affecting MMP-2 levels.
- These findings highlight distinct mechanisms by which IFN and anti-CD44 antibody can impede glioma cell invasion.
