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Molecular beacon-decorated polymethylmethacrylate core-shell fluorescent nanoparticles for the detection of survivin mRNA in human cancer cells
閱讀：896 發布時間：2018-1-30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.