Inhibition of matrix metalloproteinase-14 in osteosarcoma cells by clodronate
| Autor: | Heikkilä, P., Teronen, O., Hirn, M.Y., Sorsa, T., Tervahartiala, T., Salo, T., Konttinen, Y.T., Halttunen, T., Moilanen, M., Hanemaaijer, R., Laitinen, M. |
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| Jazyk: | angličtina |
| Rok vydání: | 2003 |
| Předmět: |
Matrix Metalloproteinases
Membrane-Associated Fluorescent Antibody Technique Physiological Sciences phorbol 13 acetate 12 myristate MT1-MMP osteosarcoma Matrix Metalloproteinase 13 Tumor Cells Cultured Bisphosphonate Humans controlled study human Collagenases RNA Messenger Enzyme Inhibitors Biology enzyme inhibition protein expression Inhibition cancer cell Enzyme Precursors Binding Sites MMP-2 human cell article Metalloendopeptidases enzyme activation Blotting Northern Recombinant Proteins cell activation enzyme activity clodronic acid priority journal Gelatinases Culture Media Conditioned immunohistochemistry protein degradation cytotoxicity Northern blotting Matrix Metalloproteinase 2 Tetradecanoylphorbol Acetate matrix metalloproteinase 14 down regulation |
| Zdroj: | Journal of Surgical Research, 1, 111, 45-52 |
| Popis: | Background. Bisphosphonates reduce the bone metastasis formation and angiogenesis but the exact molecular mechanisms involved are unclear. Progelatinase A (proMMP-2; 78 KDa) is activated up during the tumor spread and metastasis by a cell surface-associated matrix metalloproteinase (membrane-type matrix metalloproteinase [MT1-MMP] or MMP-14). Material and methods. We evaluated the effects of a bisphosphonate (clodronate) on MT1-MMP mRNA expression and protein production, catalytic activity and proteolytic activation of proMMP-2 by cultured human MG-63 osteosarcoma cells. Results. Clodronate, at therapeutically attainable noncytotoxic concentrations, dose-dependently inhibited phorbol myristic acetate (PMA)-induced proteolytic activation of proMMP-2 by human MG-63 osteosarcoma cells. Clodronate also downregulated the PMA-induced expression of MT1-MMP mRNA and protein production in human MG-63 osteosarcoma cells, as evidenced by Northern analysis and fluorescent immunohistochemistry. Furthermore, clodronate inhibited directly and dose-dependently MT1-MMP activity, and the MT1-MMP inhibition by clodronate was reduced in the presence of an increased (5 mM) Ca2+ concentrations when compared to physiological (1 mM) Ca2+ concentrations. Conclusion. We conclude that (1) the extracellular/cell-associated mechanism of bisphosphonate involves inhibition of MT1-MMP catalytic activity eventually by chelation, and that (2) intracellular mechanism involves downregulation of induced MT1-MMP mRNA and protein expression. The inhibition and downregulation of MT1-MMP by clodronate can be related to their ability to reduce MG-63 osteosarcoma cell invasion and spread. These findings may, at least in part, explain at molecular level the antitumor and antibone resorption activities of clodronate observed in clinical studies. © 2003 Elsevier Inc. All rights reserved. |
| Databáze: | OpenAIRE |
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