Downregulation of MMP-9 in ERK-mutated stable transfectants inhibits glioma invasion in vitro.
We previously showed that enhanced expression of MMP-9, an endopeptidase that digests basement-membrane type IV collagen, is related to tumor progression in vitro and in vivo; antisense-MMP-9 stably transfected clones were less invasive than untransfected parental cells and did not form tumors in nude mice. In this study, we examined the role of ERK-1 in the regulation of MMP-9 production and the invasive behavior of the human glioblastoma cell line SNB19, in which ERK1 is constitutively activated. SNB19 cells were stably transfected with mt-ERK, a vector encoding ERK-1 cDNA in which the conserved lysine at codon 71 was changed to arginine, thus impairing the catalytic efficiency of this enzyme. Gelatin zymography showed reduced levels of MMP-9 in the mt-ERK-transfected cell lines relative to those in vector-transfected and parental control cells. Reductions in MMP-9 protein mRNA levels were also detected in the mt-ERK-transfected cells by Western and Northern blotting. The mt-ERK-transfected cells were much less invasive than parental or vector control cells in a Matrigel invasion assay and in a spheroid coculture assay. Thus an ERK-dependent signaling pathway seems to regulate MMP-9 mediated glioma invasion in SNB19 cells; interfering with this pathway could be developed into a therapeutic approach, which aims at a reduction of cancer cell invasion.
Lakka
Sajani S
SS
Division of Cancer Biology, Department of Biomedical and Therapeutic Sciences, University of Illinois College of Medicine at Peoria, 61656, USA.
Jasti
Sushma L
SL
Gondi
Christopher
C
Boyd
Douglas
D
Chandrasekar
Nirmala
N
Dinh
Dzung H
DH
Olivero
William C
WC
Gujrati
Meena
M
Rao
Jasti S
JS
D015152:Blotting, Northern; D015153:Blotting, Western; D001932:Brain Neoplasms / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D002455:Cell Division; D003094:Collagen; D015536:Down-Regulation; D004338:Drug Combinations; D024202:Electrophoretic Mobility Shift Assay; D005780:Gelatin / Q000378:metabolism; D005910:Glioma / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D006801:Humans; D007797:Laminin; D020780:Matrix Metalloproteinase 9 / Q000235:genetics / Q000378:metabolism*; D018613:Microscopy, Confocal; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases / Q000235:genetics* / Q000378:metabolism; D009154:Mutation / Q000235:genetics*; D016328:NF-kappa B / Q000378:metabolism; D009361:Neoplasm Invasiveness; D011401:Promoter Regions, Genetic; D011509:Proteoglycans; D012333:RNA, Messenger / Q000378:metabolism; D015398:Signal Transduction; D018874:Spheroids, Cellular / Q000473:pathology; D018808:Transcription Factor AP-1 / Q000378:metabolism; D015533:Transcriptional Activation; D014162:Transfection; D014407:Tumor Cells, Cultured
D004338:Drug Combinations; D007797:Laminin; D016328:NF-kappa B; D011509:Proteoglycans; D012333:RNA, Messenger; D018808:Transcription Factor AP-1; C059485:matrigel; D005780:Gelatin; D003094:Collagen; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases; D020780:Matrix Metalloproteinase 9