Effect of mucin 1 aptamer conjugated nanoparticles containing docetaxel and c-Met siRNA in metastatic breast cancer cells (SKBR3)

Abstract

Introduction: Combination therapy of metastatic breast cancer with docetaxel and trastuzumab has been approved in clinic. Overexpression of cMet is one of the resistance mechanisms to trastuzumab. Chitosan nanoparticles are suitable carriers for siRNA and docetaxel delivery. MUC1 is a tumor associated antigen. Thus, anti-MUC1 aptamer can be suitable for targeted delivery of nanocarriers. Objectives: Chitosan nanoparticles were loaded with docetaxel and cMet siRNA and conjugated with MUC1 aptamer. After evaluation of pharmaceutical properties, their cellular and molecular effects in combination with trastuzumab were assessed in SKBR3 cells. Material and methods: After preparation of chitosan nanoparticles, size, zeta potential and PDI were assessed and pharmaceutical evaluations were done. Cellular uptake of nanoparticles was evaluated by immunofluorescence microscopy. For confirmation of the gene silencing, Immunoblotting was used. MTT assay was used for evaluation of cell viability. For studying of the effects of different pharmaceutical groups on genes expression, apoptosis and migration, Real-time PCR, Annexin V assay and scratch test were used, respectively. Results: Pharmaceutical evaluations represented the proper shape, size, zeta potential, loading of siRNA and conjugation of docetaxel. Nanoparticles were stable in serum upto 8 hours and in the presence of heparin didn’t release siRNA. The highest cellular uptake was observed in aptamer conjugated nanoparticles. The cMet silencing was observed after 48 hours of treatment with siRNA loaded nanoparticles. The most decreasing in cell viability, genes expression, and migration and most increasing in apoptosis were observed in co-treatment of trastuzumab and complete nanodrug. Conclusion: Targeted co-delivery of docetaxel and cMet siRNA with trastuzumab can lead to the decreasing cell viability, malignant genes expression and migration and also increasing of the apoptosis in the metastatic tumor cells. In addition, further animal model experiments are needed for validation of this functionalized nanodrug