Molecular beacon-decorated polymethylmethacrylate core-shell fluorescent nanoparticles for the detection of survivin mRNA in human cancer cells

• Published in: Biosensors & bioelectronics 2017-02, Vol.88, p.15-24
• Main Authors: Adinolfi, Barbara, Pellegrino, Mario, Giannetti, Ambra, Tombelli, Sara, Trono, Cosimo, Sotgiu, Giovanna, Varchi, Greta, Ballestri, Marco, Posati, Tamara, Carpi, Sara, Nieri, Paola, Baldini, Francesco
• Format: Article
• Language: English
• Subjects: A549 Cells; A549 human lung adenocarcinoma epithelial cells; Biosensing Techniques - methods; Bright spots; Cancer; Cellular up-take; Chromium; Copolymerization; Core-shell polymethylmethacrylate nanoparticles; Fluorescence; Fluorescent Dyes - chemistry; Humans; Inhibitor of Apoptosis Proteins - genetics; Molecular beacon; Nanoparticles; Nanoparticles - chemistry; Nanoparticles - ultrastructure; Nanostructure; Optical Imaging - methods; Polymethyl Methacrylate - chemistry; Polymethyl methacrylates; RNA Probes - chemistry; RNA Probes - genetics; RNA, Messenger - analysis; RNA, Messenger - genetics; Sensing in cell; Spectrometry, Fluorescence - methods; Survivin
• ISSN: 0956-5663; 1873-4235
• DOI: 10.1016/j.bios.2016.05.102

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. •We reported a specific molecular beacon for the survivin mRNA detection in living cells.•Polymethylmethacrylate Core-shell Fluorescent Nanoparticles promote the internalization of the molecular beacon in Human cells.•The nanocomplex (MB@PMMA-NPs) is a promising strategy to reduce cancer cell proliferation avoiding detectable consequences on healthy cells.