Abstract
BackgroundOverexpression of the RAN GTP (RAN) gene has been shown to be linked to metastatic activity of MDA-MB231 human breast cancer cells by increasing Ras/MEK/ERK and PI3K/Akt/mTORC1 signalling. The aim of this study was to investigate the potential of polymeric nanoparticles to deliver two novel shRNA sequences, targeted against the RAN gene, to MDA-MB231 cells grown in culture and to assess their effects in a range of biological assays.MethodsBiodegradable PLGA nanoparticles, loaded with shRNA-1 and shRNA-4, were fabricated using a double emulsion solvent evaporation technique and characterised for size, zeta potential and polydispersity index before testing on the MDA-MB231 cell line in a range of assays including cell viability, migration, invasion and gene knock down.ResultsshRNA-loaded nanoparticles were successfully fabricated and delivered to MDA-MB231 cells in culture, where they effectively released their payload, causing a decrease in both cell invasion and cell migration by knocking down RAN gene expression.ConclusionResults indicate the anti-RAN shRNA-loaded nanoparticles deliver and release biological payload to MDA-MB231 cells in culture. This works paves the way for further investigations into the possible use of anti-RAN shRNA-loaded NP formulations for the treatment of breast cancer in vivo.
Highlights
Genetic instability in cellular function has a significant bearing on the transition from normal to cancerous growth patterns [1]
Of particular interest to our group is the overexpression of the RAN gene in breast cancer cells, which has attracted much attention, with results demonstrating its clear role in the metastatic process [4]
Cell culture studies were performed on the MDA-MB231 breast cancer cell line, which was obtained from SigmaAldrich Ltd. (Dorset, UK)
Summary
Genetic instability in cellular function has a significant bearing on the transition from normal to cancerous growth patterns [1]. It is widely accepted that metastatic disease is associated with high rates of mortality observed in cancer patients and inhibiting metastasis is, subject to intense investigation [3] These investigations focus on the function and mutation in specific genes [3]. Of particular interest to our group is the overexpression of the RAN gene in breast cancer cells, which has attracted much attention, with results demonstrating its clear role in the metastatic process [4]. This functional loss of cell cycle machinery causes cells to acquire aggressive and invasive characteristics, enabling invasion at distant sites. Results shRNA-loaded nanoparticles were successfully fabricated and delivered to MDA-MB231 cells in culture, where they effectively released their payload, causing a decrease in both cell invasion and cell migration by knocking down RAN gene expression
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