Vpliv monoklonskih protiteles proti katepsinu B in proteinaznih inhibitorjev na proliferacijo, invazijo in angiogenezo tumorskih celic in vitro : magistrsko delo = The effects of anti-cathepsin B monoclonal antibodies and proteinaze inhibitors on tumor cell proliferation, invasion and angiogenesis in vitro : master thesis
Premzl, Aleš, 1971-
Lysosomal cysteine proteinases cathepsins B and L and aspartic proteinase cathepsin D are associated with the processes of tumor progression. The proteolytic activity of cathepsin B has been ...suggested to facilitate degradation of extracellular matrix, leading to invasion, metastasis and tumorangiogenesis. Several natural and synthetic proteinase inhibitors have not provided, expected antitumor activity in vivo, probably due to insufficient specificity and high toxicity. Monoclonal antibodies neutralizingenzyme activity are more specific and less toxic compared to the inhibitors and may serve as alternative agents. Fusing mouse splenocytes and mieloma cells we prepared neutralizing monoclonal antibody, which is capable to bind an epitope near the active site of cathepsin B, and to inhibit in vitro its proteolytic activity against fluorogenic substrates Z-Arg-Arg-AMC and BODIPY FL casein. Its inhibitory effect was tested also on tumor cell viability, proliferation and invasion and compared to several natural and synthetic inhibitors of cysteine and aspartic proteinases. For this purpose MCF-l0A neoT transformed human breast epithelial cells were used, expressing cathepsins B, L and D as detected by immunofluorescence microscopy. Monoclonalantibody significantly decreased invasion (47.3 +- 0.3 % at 0.5 microM, whereas no effect on cell viability and proliferation was detected in the same concentration range. Among cysteine proteinase inhibitors, the most effective was chicken cystatin (82.8 ` 1.0%, 2 N,M), the next was E-64 (56.5 f1.3%, 10 N,M). Both generai inhibitors of cysteine proteinases showed higher inhibition compared to cathepsin L specific inhibitor CLIK-148, (32.7 microM 1.5%, 10 microM. (Abstract truncated at 2000 characters)
Type of material - master's thesis
Publication and manufacture - Ljubljana : [A. Premzl], 2001
National Institute of Biology and BF, Department of Biology, Ljubljana
BF oddelek za biologijo mg 616-006 PREMZL A. Vpliv IN: 0035426
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