Polymethylmethacrylate nanoparticles as carrier of an oligodeoxynucleotide molecular beacon specific for survivin mRNA in A549 human lung adenocarcinoma epithelial cells

摘要

In cancer research, the use of antisense oligonucleotide molecular beacons, able to generate a fluorescent signal when they hybridize with their target mRNA, may represent an innovative strategy that conjugates the ability of sensing specific mRNA with the pharmacological silencing activity, preventing the overexpression of proteins associated to cancer development. In cancer context, this approach minimizes the non-specific toxicity and addresses the therapy mainly towards the tumor cells by using effective delivery systems. The aim of this work was to investigate the ability of polymethylmethacrylate nanoparticles (PMMA-NPs) to act as carrier of a theranostic agent, an oligonucleotide molecular beacon (MB) targeting survivin mRNA in A549 human lung adenocarcinoma epithelial cells. Moreover, this paper highlights the need for having an appropriate healthy control in in-vitro experiments. This is a problem widely discussed and felt by the scientific community and often represents a limit recognized in many experimental approaches. In particular, the survivin-MB was firstly characterized in solution in order to verify its functionality and then the PMMA-NPs ability to promote the MB internalization was verified in A549 cells by confocal microscopy. Confluent Human Dermal Fibroblasts from adult (HDFa) were used as healthy control. The results obtained allow us to assess that PMMA-NPs promote the survivin-MB cellular up-take and that the use of 10 μg/mL PMMA-NPs as carrier for survivin-MB for 1h 30 mins might be a promising strategy to reduce cancer cell proliferation avoiding detectable consequences on the healthy cells.