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Tissue inhibitor of metalloproteinase 3 suppresses tumor angiogenesis in matrix metalloproteinase 2-down-regulated lung cancer.
- Source :
-
Cancer research [Cancer Res] 2008 Jun 15; Vol. 68 (12), pp. 4736-45. - Publication Year :
- 2008
-
Abstract
- Matrix metalloproteinase-2 (MMP-2) expression is often up-regulated in advanced cancers and known to play an important role in tumor angiogenesis. We previously showed that adenoviral-mediated delivery of siRNA for MMP-2 (Ad-MMP-2-Si) inhibited lung cancer growth, angiogenesis, and metastasis. In this study, we investigated the signaling mechanisms involved in Ad-MMP-2-Si-mediated inhibition of angiogenesis. Ad-MMP-2-Si treatment inhibited neovascularization in vivo as determined by mouse dorsal air sac model, and conditioned medium from Ad-MMP-2-Si-infected A549 lung cancer cells (Ad-MMP-2-Si-CM) inhibited endothelial tube formation in vitro. Ad-MMP-2-Si-CM decreased proliferation as determined by Ki-67 immunofluorescence and induced apoptosis in endothelial cells as determined by terminal deoxynucleotidyl-transferase-mediated dUTP nick-end labeling (TUNEL) assay. Furthermore, Ad-MMP-2-Si-CM inhibited AKT phosphorylation and induced phosphorylation of extracellular signal-regulated kinase (ERK)/mitogen-activated protein kinase in endothelial cells. Overexpression of constitutively active AKT reversed the Ad-MMP-2-Si-CM-mediated inhibition of tube formation and induction of ERK phosphorylation. Conversely, Ad-MMP-2-Si-CM induced tissue inhibitor of metalloproteinase (TIMP) 3 expression, and the interaction of vascular endothelial growth factor 2 and TIMP-3 was determined by coimmunoprecipitation experiments. TIMP-3 induction was mediated by ERK activation. In addition, electrophoretic mobility shift and chromatin immunoprecipitation assays show that Sp1 transcription factor mediated Ad-MMP-2-Si-CM-stimulated increase of TIMP-3. Vasculature destruction was confirmed with colocalization studies with TUNEL and an endothelial marker, CD31, in tumor sections of Ad-MMP-2-Si-treated mice. Our data collectively suggest that MMP-2 inhibition induces endothelial apoptosis in vivo and inhibits endothelial tube formation. These experiments provide the first evidence that inhibition of p-AKT and induction of p-ERK1/2 are crucial events in the induction of TIMP-3-mediated endothelial apoptosis in MMP-2 inhibited lung tumors.
- Subjects :
- Adenoviridae genetics
Animals
Apoptosis
Cell Proliferation
Chromatin Immunoprecipitation
Culture Media, Conditioned pharmacology
Down-Regulation
Electrophoretic Mobility Shift Assay
Endothelium, Vascular
Extracellular Signal-Regulated MAP Kinases metabolism
Fluorescent Antibody Technique
Humans
Immunoblotting
Immunoprecipitation
Matrix Metalloproteinase 2 genetics
Mice
Mice, SCID
Mitogen-Activated Protein Kinases metabolism
Phosphorylation
Proto-Oncogene Proteins c-akt metabolism
RNA, Small Interfering pharmacology
Signal Transduction
Tumor Cells, Cultured
Vascular Endothelial Growth Factor A metabolism
Xenograft Model Antitumor Assays
Lung Neoplasms blood supply
Lung Neoplasms enzymology
Matrix Metalloproteinase 2 metabolism
Neovascularization, Pathologic prevention & control
Tissue Inhibitor of Metalloproteinase-3 pharmacology
Subjects
Details
- Language :
- English
- ISSN :
- 1538-7445
- Volume :
- 68
- Issue :
- 12
- Database :
- MEDLINE
- Journal :
- Cancer research
- Publication Type :
- Academic Journal
- Accession number :
- 18559520
- Full Text :
- https://doi.org/10.1158/0008-5472.CAN-07-6612