Molecular beacon-decorated polymethylmethacrylate core-shell fluorescent nanoparticles for the detection of survivin mRNA in human cancer cells.
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 60nm, 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±0.20µm2), 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±0.16µm2). 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.
Istituto di Fisica Applicata "Nello Carrara", Consiglio Nazionale delle Ricerche, Via Madonna del Piano 10, 50019 Sesto Fiorentino (Fi), Italy. Electronic address: b.adinolfi@ifac.cnr.it.
Barbara
B
Adinolfi
Dipartimento di Ricerca Traslazionale e delle Nuove Tecnologie in Medicina e Chirurgia, Università di Pisa, Via Savi 10, 56126 Pisa, Italy. Electronic address: mario.pellegrino@unipi.it.
Mario
M
Pellegrino
Istituto di Fisica Applicata "Nello Carrara", Consiglio Nazionale delle Ricerche, Via Madonna del Piano 10, 50019 Sesto Fiorentino (Fi), Italy. Electronic address: a.giannetti@ifac.cnr.it.
Ambra
A
Giannetti
Istituto di Fisica Applicata "Nello Carrara", Consiglio Nazionale delle Ricerche, Via Madonna del Piano 10, 50019 Sesto Fiorentino (Fi), Italy. Electronic address: s.tombelli@ifac.cnr.it.
Sara
S
Tombelli
Istituto di Fisica Applicata "Nello Carrara", Consiglio Nazionale delle Ricerche, Via Madonna del Piano 10, 50019 Sesto Fiorentino (Fi), Italy. Electronic address: c.trono@ifac.cnr.it.
Cosimo
C
Trono
Istituto per la Sintesi Organica e la Fotoreattività, Consiglio Nazionale delle Ricerche, Via P. Gobetti 101, 40129 Bologna, Italy. Electronic address: giovanna.sotgiu@isof.cnr.it.
Giovanna
G
Sotgiu
Istituto per la Sintesi Organica e la Fotoreattività, Consiglio Nazionale delle Ricerche, Via P. Gobetti 101, 40129 Bologna, Italy. Electronic address: greta.varchi@isof.cnr.it.
Greta
G
Varchi
Istituto per la Sintesi Organica e la Fotoreattività, Consiglio Nazionale delle Ricerche, Via P. Gobetti 101, 40129 Bologna, Italy. Electronic address: marco.ballestri@isof.cnr.it.
Marco
M
Ballestri
Istituto per la Sintesi Organica e la Fotoreattività, Consiglio Nazionale delle Ricerche, Via P. Gobetti 101, 40129 Bologna, Italy. Electronic address: tamara.posati@isof.cnr.it.
Tamara
T
Posati
Dipartimento di Farmacia, Università di Pisa, Via Bonanno Pisano 6, 56126 Pisa, Italy. Electronic address: sara.carpi@for.unipi.it.
Sara
S
Carpi
Dipartimento di Farmacia, Università di Pisa, Via Bonanno Pisano 6, 56126 Pisa, Italy. Electronic address: paola.nieri@farm.unipi.it.
Paola
P
Nieri
Istituto di Fisica Applicata "Nello Carrara", Consiglio Nazionale delle Ricerche, Via Madonna del Piano 10, 50019 Sesto Fiorentino (Fi), Italy. Electronic address: f.baldini@ifac.cnr.it.
Francesco
F
Baldini
D000072283:A549 Cells; D015374:Biosensing Techniques / Q000379:methods; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000235:genetics*; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D061848:Optical Imaging / Q000379:methods; D019904:Polymethyl Methacrylate / Q000737:chemistry*; D015347:RNA Probes / Q000737:chemistry* / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics*; D013050:Spectrometry, Fluorescence / Q000379:methods; D000077022:Survivin
C107489:BIRC5 protein, human; D005456:Fluorescent Dyes; D051034:Inhibitor of Apoptosis Proteins; D015347:RNA Probes; D012333:RNA, Messenger; D000077022:Survivin; D019904:Polymethyl Methacrylate
A549 human lung adenocarcinoma epithelial cells; Cellular up-take; Core-shell polymethylmethacrylate nanoparticles; Molecular beacon; Sensing in cell; Survivin