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Suppression of MMP-2 attenuates TNF-α induced NF-κB activation and leads to JNK mediated cell death in glioma.
- Source :
-
PloS one [PLoS One] 2011 May 04; Vol. 6 (5), pp. e19341. Date of Electronic Publication: 2011 May 04. - Publication Year :
- 2011
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Abstract
- Background: Abrogation of apoptosis for prolonged cell survival is essential in cancer progression. In our previous studies, we showed the MMP-2 downregulation induced apoptosis in cancer cell lines. Here, we attempt to investigate the exact molecular mechanism of how MMP-2 depletion leads to apoptosis in glioma xenograft cell lines.<br />Methodology/principal Findings: MMP-2 transcriptional suppression by MMP-2siRNA (pM) induces apoptosis associated with PARP, caspase-8 and -3 cleavage in human glioma xenograft cells 4910 and 5310. Western blotting and cytokine array showed significant decrease in the cellular and secreted levels of TNF-α with concomitant reduction in TNFR1, TRADD, TRAF2, RIP, IKKβ and pIκBα expression levels resulting in inhibition of p65 phosphorylation and nuclear translocation in pM-treated cells when compared to mock and pSV controls. In addition MMP-2 suppression led to elevated Fas-L, Fas and FADD expression levels along with increased p38 and JNK phosphorylation. The JNK-activity assay showed prolonged JNK activation in pM-transfected cells. Specific inhibition of p38 with SB203580 did not show any effect whereas inhibition of JNK phosphorylation with SP600125 notably reversed pM-induced cleavage of PARP, caspase-8 and -3, demonstrating a significant role of JNK in pM-induced cell death. Supplementation of rhMMP-2 counteracted the effect of pM by remarkably elevating TNF-α, TRADD, IKKβ and pIκBα expression and decreasing FADD, Fas-L, and phospho-JNK levels. The EMSA analysis indicated significant reversal of pM-inhibited NF-κB activity by rhMMP-2 treatment which rescued cells from pM-induced cell death. In vivo studies indicated that pM treatment diminished intracranial tumor growth and the immuno histochemical analysis showed decreased phospho-p65 and enhanced phospho-JNK levels that correlated with increased TUNEL-positive apoptotic cells in pM-treated tumor sections.<br />Conclusion/significance: In summary, our study implies a role of MMP-2 in the regulation of TNF-α mediated constitutive NF-κB activation and Fas-mediated JNK mediated apoptosis in glioma xenograft cells in vitro and in vivo.
- Subjects :
- Animals
Anthracenes pharmacology
Apoptosis genetics
Blotting, Western
Cell Line, Tumor
Electrophoresis, Polyacrylamide Gel
Electrophoretic Mobility Shift Assay
Enzyme Inhibitors pharmacology
Flow Cytometry
Glioma genetics
Humans
Imidazoles pharmacology
Immunohistochemistry
Immunoprecipitation
In Situ Nick-End Labeling
JNK Mitogen-Activated Protein Kinases antagonists & inhibitors
Matrix Metalloproteinase 2 genetics
Mice
Pyridines pharmacology
RNA, Small Interfering
Reverse Transcriptase Polymerase Chain Reaction
Tissue Inhibitor of Metalloproteinase-3 genetics
Tissue Inhibitor of Metalloproteinase-3 metabolism
Apoptosis drug effects
Glioma metabolism
Glioma therapy
JNK Mitogen-Activated Protein Kinases metabolism
Matrix Metalloproteinase 2 metabolism
NF-kappa B metabolism
Tumor Necrosis Factor-alpha pharmacology
Subjects
Details
- Language :
- English
- ISSN :
- 1932-6203
- Volume :
- 6
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- PloS one
- Publication Type :
- Academic Journal
- Accession number :
- 21573233
- Full Text :
- https://doi.org/10.1371/journal.pone.0019341