Poly(lactide-co-glycolide) nanoparticles as a carrier system for delivering cysteine protease inhibitor cystatin into tumor cells
| Autor: | Aleš Premzl, Valentina Zavašnik-Bergant, Mateja Cegnar, Janko Kos, Julijana Kristl |
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| Rok vydání: | 2004 |
| Předmět: |
Proteases
Cell Survival Biology Cysteine Proteinase Inhibitors urologic and male genital diseases Cathepsin B chemistry.chemical_compound Cell Line Tumor Neoplasms Animals Humans Particle Size Polyglactin 910 Cathepsin Drug Carriers technology industry and agriculture Biological activity Cell Biology Cysteine protease Cystatins female genital diseases and pregnancy complications PLGA Biochemistry chemistry Cystatin Chickens Intracellular Cysteine |
| Zdroj: | Experimental cell research. 301(2) |
| ISSN: | 0014-4827 |
| Popis: | 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 μg/ml were cytotoxic towards MCF-10A neoT cells, but not free cystatin at concentrations up to 5 μM. 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 inhibited cathepsin 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 against intracellular targets. |
| Databáze: | OpenAIRE |
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