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Molecularbeacon-decoratedpolymethylmethacrylatecore-shellfluorescentnanoparticlesforthedetectionofsurvivinmRNAinhumancancercells

时间:2018-01-30      阅读:787

作者 Barbara Adinolfia. Mario Pellegrinob. Ambra Giannettia. Sara Tombellia. Cosimo Tronoa. Giovanna Sotgiuc. Greta Varchic. Marco Ballestric. Tamara Posatic. Sara Carpid. Paola Nierid. Francesco Baldinia.

a

Istituto di Fisica Applicata “Nello Carrara”, Consiglio Nazionale delle Ricerche, Via Madonna del Pia0, 50019 Sesto Fiorentino (Fi), Italy

b

Dipartimento di Ricerca Traslazionale e delle Nuove Tecnologie in Medicina e Chirurgia, Università di Pisa, Via Savi 10, 56126 Pisa, Italy

c

Istituto per la Sintesi Organica e la Fotoreattività, Consiglio Nazionale delle Ricerche, Via P. Gobetti 101, 40129 Bologna, Italy

d

Dipartimento di Farmacia, Università di Pisa, Via Bonanno Pisano 6, 56126 Pisa, Italy

 

摘要:One of the main goals of nanomedicine in cancer is the development of effective drug delivery systems, primarily nanoparticles.

 

Survivin, an overexpressed anti-apoptotic protein in cancer, represents a pharmacological target for therapy and a Molecular Beacon (MB) specific for survivin mRNA is available. In this study, the ability of polymethylmethacrylate nanoparticles (PMMA-NPs) to promote survivin MB uptake in human A549 cells was investigated.

 

Fluorescent and positively charged core PMMA-NPs of nearly 60 nm, obtained through an emulsion co-polymerization reaction, and the MB alone were evaluated in solution, for their analytical characterization; then, the MB specificity and functionality were verified after adsorption onto the PMMA-NPs.

The carrier ability of PMMA-NPs in A549 was examined by confocal microscopy. With the optimized protocol, a hardly detectable fluorescent signal was obtained after incubation of the cells with the MB alone (fluorescent spots per cell of 1.90±0.40 with a mean area of 1.04±0.20 µm2), while bright fluorescent spots inside the cells were evident by using the MB loaded onto the PMMA-NPs. (27.50±2.30 fluorescent spots per cell with a mean area of 2.35±0.16 µm2). These results demonstrate the ability of the PMMA-NPs to promote the survivin-MB internalization, suggesting that this complex might represent a promising strategy for intracellular sensing and for the reduction of cancer cell proliferation.

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