Activation of caspase-dependent apoptosis by intracellular delivery of cytochrome c-based nanoparticles

Journal of Nanobiotechnology

Research

Yamixa Delgado¹  Moraima Morales-Cruz¹  Myraida Morales²  Jessica Méndez³  Cindy M Figueroa³  Tania González-Robles³  Kai Griebenow³  Anna Molina³ 

[1] Department of Biology, University of Puerto Rico, Río Piedras Campus, 00931, San Juan, PR, USA;Department of Graduate Studies, University of Puerto Rico, Río Piedras Campus, 00931, San Juan, PR, USA;Departments of Chemistry, University of Puerto Rico, Río Piedras Campus, 00931, San Juan, PR, USA;

关键词: Drug delivery;    Protein nanoparticles;    PLGA;    Passive targeting;    Triggered release;   

DOI  :  10.1186/s12951-014-0033-9

 received in 2014-05-30, accepted in 2014-08-23,  发布年份 2014

来源: Springer

【 摘 要 】

BackgroundCytochrome c is an essential mediator of apoptosis when it is released from the mitochondria to the cytoplasm. This process normally takes place in response to DNA damage, but in many cancer cells (i.e., cancer stem cells) it is disabled due to various mechanisms. However, it has been demonstrated that the targeted delivery of Cytochrome c directly to the cytoplasm of cancer cells selective initiates apoptosis in many cancer cells. In this work we designed a novel nano-sized smart Cytochrome c drug delivery system to induce apoptosis in cancer cells upon delivery.ResultsCytochrome c was precipitated with a solvent-displacement method to obtain protein nanoparticles. The size of the Cytochrome c nanoparticles obtained was 100-300nm in diameter depending on the conditions used, indicating good potential to passively target tumors by the Enhanced Permeability and Retention effect. The surface of Cytochrome c nanoparticles was decorated with poly (lactic-co-glycolic) acid-SH via the linker succinimidyl 3-(2-pyridyldithio) propionate to prevent premature dissolution during delivery. The linker connecting the polymer to the protein nanoparticle contained a disulfide bond thus allowing polymer shedding and subsequent Cytochrome c release under intracellular reducing conditions. A cell-free caspase-3 assay revealed more than 80% of relative caspase activation by Cytochrome c after nanoprecipitation and polymer modification when compared to native Cytochrome c. Incubation of HeLa cells with the Cytochrome c based-nanoparticles showed significant reduction in cell viability after 6hours while native Cytochrome c showed none. Confocal microscopy confirmed the induction of apoptosis in HeLa cells when they were stained with 4’,6-diamidino-2-phenylindole and propidium iodide after incubation with the Cytochrome c-based nanoparticles.ConclusionsOur results demonstrate that the coating with a hydrophobic polymer stabilizes Cytochrome c nanoparticles allowing for their delivery to the cytoplasm of target cells. After smart release of Cytochrome c into the cytoplasm, it induced programmed cell death.

【 授权许可】

Unknown   
© Morales-Cruz et al.; licensee BioMed Central Ltd. 2014. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.