Rat liver myofibroblasts (MFB) were isolated by repeated passaging of nonparenchymal liver cell fraction. They were cultured on polystyrene Petri dishes, on fibrin or on type I collagen gels for 5 days. Quantitative RT-PCR, Western blotting, zymography and immunocytochemistry were used to study differences in cell morphology and protein expression. MFB were large and spread on plastic substrate, with prominent alpha-smooth muscle (alpha-SMA) fibres. They turned much smaller and elongated on collagen which was accompanied by the rearrangement of the cytoskeleton and a decrease in alpha-SMA and beta-actin content. Collagen gel induced the expression of a group of metalloproteinases (MMP-2, -3, -9, -13), on mRNA and protein level which resulted in the degradation of the gel. This response was accompanied by changes in the mRNA expression of cytokines of TGF-beta family, CTGF and interleukin-6, as well as of osteopontin and thrombospondin-2 that are involved in metalloproteinases (MMPs) regulation. The expression of MMPs substrates, collagen types I, IV and XII did not change or decreased. The effects of fibrin gels on MFB were milder than those of collagen. MFB assumed to deposit collagen and other ECM components in fibrotic liver, besides hepatic stellate cells, also possess a great collagenolytic potential.
- MeSH
- aktiny metabolismus MeSH
- biologické markery metabolismus MeSH
- časové faktory MeSH
- cytokiny metabolismus MeSH
- cytoskelet enzymologie MeSH
- fibrin metabolismus MeSH
- imunohistochemie MeSH
- játra cytologie enzymologie MeSH
- kolagen typu I metabolismus MeSH
- kolagenasy genetika metabolismus MeSH
- krysa rodu rattus MeSH
- kultivované buňky MeSH
- messenger RNA metabolismus MeSH
- myofibroblasty enzymologie MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- potkani Sprague-Dawley MeSH
- separace buněk metody MeSH
- tvar buňky MeSH
- western blotting MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The newly established breast cancer cell line G3S1, derived from EM-G3 breast cancer progenitors, was analyzed for functional changes related to neoplastic progression manifested by elevated invasiveness and enhanced capability to degrade gelatin. Degradation of gelatin and invasiveness of G3S1 cells was found to be dependent on the activity of matrix proteinases and actin cytoskeletal dynamics. Therefore, the expression and activity of these proteases was compared in G3S1 and EM-G3 cells. Despite enhanced capability of G3S1 cells to degrade gelatin, these cells exhibited lower levels of secreted extracellular matrix degrading proteases than parental EM-G3 cells. However, the expression of membrane-bound MT1-MMP was strongly elevated in G3S1 cells. While the degradation of gelatin was associated with invadopodia-like structures in both EM-G3 and G3S1 cells, the cytoskeletal remodeling dynamics was greatly elevated in G3S1 cells, suggesting that upregulation of MT1-MMP, together with elevation of cytoskeletal remodeling dynamics can effectively cause elevated invasiveness and enhanced matrix degrading capability in G3S1 cells.
- MeSH
- aktiny metabolismus MeSH
- aprotinin farmakologie MeSH
- cytoskelet účinky léků enzymologie patologie MeSH
- dipeptidy farmakologie MeSH
- inhibitory proteas farmakologie MeSH
- invazivní růst nádoru MeSH
- leucin analogy a deriváty farmakologie MeSH
- lidé MeSH
- matrixová metaloproteinasa 14 antagonisté a inhibitory metabolismus MeSH
- metaloproteinasy secernované do matrix metabolismus MeSH
- mořské toxiny farmakologie MeSH
- nádorové buněčné linie MeSH
- nádorové kmenové buňky enzymologie patologie MeSH
- nádory prsu enzymologie patologie MeSH
- pohyb buněk účinky léků MeSH
- progrese nemoci MeSH
- pseudopodia enzymologie MeSH
- upregulace MeSH
- želatina metabolismus MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
