TY - JOUR
T1 - Polymeric nanoparticles promote endocytosis of a survivin molecular beacon
T2 - Localization and fate of nanoparticles and beacon in human A549 cells
AU - Adinolfi, Barbara
AU - Pellegrino, Mario
AU - Tombelli, Sara
AU - Trono, Cosimo
AU - Giannetti, Ambra
AU - Domenici, Claudio
AU - Varchi, Greta
AU - Sotgiu, Giovanna
AU - Ballestri, Marco
AU - Baldini, Francesco
N1 - Copyright © 2018. Published by Elsevier Inc.
PY - 2018/12/15
Y1 - 2018/12/15
N2 - Polymethylmethacrylate core-shell fluorescent nanoparticles promote, in human lung A549 cancer cells, the internalization of a molecular beacon (MB) specific for survivin mRNA, an anti-apoptotic protein overexpressed in cancer cells. Aims: To design an effective drug delivery system, the knowledge of the uptake mechanism and of the nanoparticles (NPs) and MB fate is required. Materials and methods and key findings: Experiments with dextran as marker for endocytosis showed that in the presence of NPs the number of endocytic vesicles per cell doubled and their mean size significantly (p < 0.001) increased with respect to controls in absence of NPs, indicating an involvement of NPs in the endocytotic process. By using LysoTracker™ Deep Red, as marker of lysosomes, we found that nanoparticles co-localize with lysosomes. Moreover, a cellular release of nanoparticles detected in the culture medium, suggested a role of lysosomal exocytosis in nanoparticle elimination. The MB fluorescence in proximity of the labeled Endoplasmic Reticulum was indicative that the opening of the MB occurs in proximity of its target mRNA. Significance: The results show the involvement of endocytotic pathway in the uptake of NPs, which are an appropriate delivery system capable of being eliminated by cells. Furthermore the data confirm that the MB can be considered an effective tool for the intracellular sensing.
AB - Polymethylmethacrylate core-shell fluorescent nanoparticles promote, in human lung A549 cancer cells, the internalization of a molecular beacon (MB) specific for survivin mRNA, an anti-apoptotic protein overexpressed in cancer cells. Aims: To design an effective drug delivery system, the knowledge of the uptake mechanism and of the nanoparticles (NPs) and MB fate is required. Materials and methods and key findings: Experiments with dextran as marker for endocytosis showed that in the presence of NPs the number of endocytic vesicles per cell doubled and their mean size significantly (p < 0.001) increased with respect to controls in absence of NPs, indicating an involvement of NPs in the endocytotic process. By using LysoTracker™ Deep Red, as marker of lysosomes, we found that nanoparticles co-localize with lysosomes. Moreover, a cellular release of nanoparticles detected in the culture medium, suggested a role of lysosomal exocytosis in nanoparticle elimination. The MB fluorescence in proximity of the labeled Endoplasmic Reticulum was indicative that the opening of the MB occurs in proximity of its target mRNA. Significance: The results show the involvement of endocytotic pathway in the uptake of NPs, which are an appropriate delivery system capable of being eliminated by cells. Furthermore the data confirm that the MB can be considered an effective tool for the intracellular sensing.
KW - Cancer cells
KW - Endocytosis
KW - Lysosomal exocytosis
KW - Molecular beacon
KW - Polymethylmethacrylate nanoparticles
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U2 - 10.1016/j.lfs.2018.11.007
DO - 10.1016/j.lfs.2018.11.007
M3 - Article
C2 - 30412722
AN - SCOPUS:85056213138
VL - 215
SP - 106
EP - 112
JO - Life sciences
JF - Life sciences
SN - 0024-3205
ER -