DDAB cationic lipid‑mPEG, PCL copolymer hybrid nano‑carrier synthesis and application for delivery of siRNA targeting IGF‑1R into breast cancer cells

• Autores: M. Khodaei, F. Rostamzadeh, A. H. Taromchi, H. Monirinasab, H. Fathi
• Localización: Clinical & translational oncology, ISSN 1699-048X, Vol. 23, Nº. 6, 2021, págs. 1167-1178
• Idioma: inglés
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• Resumen

  • Background and objectiveTo use siRNA molecule as a therapeutic agent in gene silencing, an efficient delivery system is necessary. Stability and clearance by reticuloendothelial of siRNA still remains the major challenges for clinical application. Herein, we could develop new lipid-polymer hybrid nanoparticles (LPHNP) as a siRNA carrier to silence insulin-like growth factor type I (IGF-1R) gene overexpression in MCF-7 human breast cancer cell line.MethodsDimethyldioctadecylammonium bromide-methoxy poly(ethylene glycol)-poly (ε-caprolactone) (DDAB-mPEG-PCL) LPHNPs were synthesized using a single step nanoprecipitation method and characterized by dynamic light scattering (DLS) and atomic force microscopy (AFM) microscope. Cytotoxicity of the nanoparticles was assessed in the MCF7 cell line using 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Desired LPHNP-siRNA complex was determined using different Nitrogen:Phosphate ratio (N/P) ratios and gel retardation. To determine the encapsulation efficiency of siRNA (%) in LPHNP, its absorbance was measured. The effect of the siRNA-LPHNP complex on IGF-1Rsilencing was assessed by reverse transcription-polymerase chain reaction (RT-PCR)Results LPHNP was synthesized using a single-step sonication method with a size below 100 nM. The viability of cells treated with hybrid nanoparticles was significantly greater than the corresponding cationic lipid (P < 0.01). As demonstrated by gel retardation assay, efficient siRNA binding to LPHNP occurred at N/P equal to 40 and siRNA encapsulation efficiency was found to be 95% ± 4 at this ratio. LPHNP-IGF-1R siRNA complex could be able to down-regulate the target more effi-ciently when it compared with the corresponded controls (P < 0.001).ConclusionIn conclusion, our results suggest that DDAB cationic lipid and mPEG-PCL copolymer hybrid nanoparticle may be a good candidate for efficient siRNA delivery.