Linked Life Data

9049977

Source:http://linkedlifedata.com/resource/pubmed/id/9049977

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
6
pubmed:dateCreated
1997-6-12
pubmed:abstractText
Remodelling of the extracellular matrix resulting from increased secretion of metalloproteinase enzymes (MMPs) is implicated in many pathological conditions, including rheumatoid arthritis, restenosis following balloon angioplasty, atherosclerosis and cancer cell invasion and metastasis. Clear definition of the normal and pathological function of individual MMPs will benefit from approaches that use gene transfer to produce increases in MMP levels that mimic those observed in pathological conditions. Similarly, gene transfer methods leading to controlled increases in levels of the tissue inhibitor of metalloproteinases (TIMPs) will help to define the function of MMPs both in vitro and in vivo. Gene transfer of TIMPs may also have therapeutic potential in pathological conditions where inhibition of MMP activity may be beneficial. We have used the adenovirus serotype 5 vector system to generate replication-deficient recombinant adenoviruses capable of expressing the MMP-9, TIMP-1 or -2 genes. High level expression is driven by the cytomegalovirus major immediate early promoter (CMV IEP). Efficient and selective over-production of each recombinant protein was shown by immunofluorescence in either rabbit smooth muscle cells (SMC) or human MCF-7 adenocarcinoma cells. High level secretion directly dependent on the multiplicity of infection (MOI) was observed for each functional transgene by gelatin zymography. Using a quantitative ELISA assay, levels of recombinant TIMP-1 were detected when SMC were infected with as low as three plaque forming units (pfu) of virus per cell in vitro. A linear increase in TIMP-1 secretion was observed up to 1000 pfu/cell of virus (0.75 ng/10(4) cells/24 h at 3 pfu/cell to 1243 ng/10(4) cells/24h at 1000 pfu/cell). Similar levels of secretion of MMP-9 and TIMP-2 were observed by Western blot analysis using the same MOI of adenovirus. Thus, recombinant adenoviruses are an efficient and flexible system for high level expression of MMPs and TIMPs and will be useful tools in the study of matrix remodelling in vivo and in vitro.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Dec
pubmed:issn
0945-053X
pubmed:author
pubmed:issnType
Print
pubmed:volume
15
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
383-95
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed-meshheading:9049977-Adenocarcinoma, pubmed-meshheading:9049977-Adenoviridae, pubmed-meshheading:9049977-Animals, pubmed-meshheading:9049977-Cell Line, pubmed-meshheading:9049977-Cloning, Molecular, pubmed-meshheading:9049977-Collagenases, pubmed-meshheading:9049977-DNA Primers, pubmed-meshheading:9049977-Genetic Vectors, pubmed-meshheading:9049977-Glycoproteins, pubmed-meshheading:9049977-Humans, pubmed-meshheading:9049977-Kidney, pubmed-meshheading:9049977-Matrix Metalloproteinase 9, pubmed-meshheading:9049977-Muscle, Smooth, pubmed-meshheading:9049977-Polymerase Chain Reaction, pubmed-meshheading:9049977-Protein Biosynthesis, pubmed-meshheading:9049977-Proteins, pubmed-meshheading:9049977-Rabbits, pubmed-meshheading:9049977-Recombinant Fusion Proteins, pubmed-meshheading:9049977-Tissue Inhibitor of Metalloproteinase-2, pubmed-meshheading:9049977-Tissue Inhibitor of Metalloproteinases, pubmed-meshheading:9049977-Transfection, pubmed-meshheading:9049977-Tumor Cells, Cultured
pubmed:year
1996
pubmed:articleTitle
Development of recombinant adenoviruses that drive high level expression of the human metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 and -2 genes: characterization of their infection into rabbit smooth muscle cells and human MCF-7 adenocarcinoma cells.
pubmed:affiliation
Department of Cardiology, University of Wales College of Medicine, Cardiff, UK.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't