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Poly(lactide-co-glycolide) nanoparticles as a carrier system for delivering cysteine protease inhibitor cystatin into tumor cellsCegnar, Mateja ...Cystatins are able to inhibit the tumor-associated activity of intracellular cysteine proteases cathepsins B and L and have been suggested as potential anticancer drugs. We have incorporated chicken ... cystatin, a model protein inhibitor of cysteine proteases, in poly(lactide-co-glycolide) (PLGA) nanoparticles (NPs) to improve its bioavailability and delivery into tumor cells. Cystatin-loaded NPs, 300-350 nm in diameter, were prepared by the double emulsion solvent diffusion method using low energy emulsification to preserve the biological activity of the protein. PLGA NPs and cystatin-loaded PLGA NPs at concentrations higher than 80 gžml were cytotoxic towards MCF-10AneoT cells, but not free cystatin at concentrations up to 5 M. To visualize the uptake of cystatin into living MCF-10A neoT cells, NPs loaded with Alexa Fluor 488-labeled cystatin were added to the culture medium. They rapidly internalized into the cells, whereas the uptake of free-labeled cystatin was very slow. Cystatin, released from the NPs, effectively inhibitedcathepsin B activity, as detected by degradation of specific Z-Arg-Arg cresyl violet substrate. In contrast, the same amount of free cystatin showed no inhibition of intracellular cathepsin B. Our results show that PLGA NPs are a useful carrier system for rapid delivery of protein inhibitors into tumor cells, enabling effective inhibition of intracellular proteolysis. The approach can be applied to other protein drugs active againstintracellular targets.Source: Experimental cell research. - ISSN 0014-4827 (Vol. 301, no. 1, 2004, str. 223-231)Type of material - article, component partPublish date - 2004Language - englishCOBISS.SI-ID - 1608305
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Database name | Field | Year |
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Links to authors' personal bibliographies | Links to information on researchers in the SICRIS system |
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Cegnar, Mateja | 22658 |
Premzl, Aleš, 1971- | 19366 |
Zavašnik-Bergant, Tina | 18286 |
Kristl, Julijana | 09032 |
Kos, Janko, 1959- | 04648 |
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and: SICRIS
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