Polymeric nanoparticles promote endocytosis of a survivin molecular beacon: localization and fate of nanoparticles and beacon in human A549 cells

Year: 2018

Authors: Adinolfi B., Pellegrino M., Tombelli S., Trono C., Giannetti A., Domenici C., Varchi G., Sotgiu G., Ballestri M., Baldini F.

Autors Affiliation: CNR, Ist Fis Applicata Nello Carrara, Via Madonna del Piano 10, I-50019 Sesto Fiorentino, FI, Italy; CNR, Ist Nazl Ott, Via G Moruzzi 1, I-56124 Pisa, PI, Italy; CNR, Ist Fisiol Clin, Via G Moruzzi 1, I-56124 Pisa, PI, Italy; CNR, Ist Sintesi Organ & Fotoreattivita, Via P Gobetti 101, I-40129 Bologna, BO, Italy.

Abstract: 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 NP 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 nanoparticles the number of endocytic vesicles per cell doubled and their mean size significantly (p<0.001) increased, indicating a promotion of endocytosis. 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 labelled 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. Journal/Review: LIFE SCIENCES

Volume: 215      Pages from: 106  to: 112

More Information: This study was supported by the National Flagship Project NANOMAX ENCODER. S. Tombelli was supported by the National project “Improving therapy for breast cancer and melanoma by transcriptome-methylome profiling, integrative network inference, and design of novel theranostic tools” funded by the Istituto Toscano Tumori (ITT).
KeyWords: Polymethylmethacrylate nanoparticles, Endocytosis, Lysosomal exocytosis, Molecular beacon, Cancer cells
DOI: 10.1016/j.lfs.2018.11.007

ImpactFactor: 3.448
Citations: 9
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