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

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Abstract

Background and objective

To 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.

Methods

Dimethyldioctadecylammonium 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-1R silencing 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 efficiently when it compared with the corresponded controls (P < 0.001).

Conclusion

In conclusion, our results suggest that DDAB cationic lipid and mPEG-PCL copolymer hybrid nanoparticle may be a good candidate for efficient siRNA delivery.

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Abbreviations

LPHNPs:

Lipid polymer hybrid nanoparticles

PEG:

Poly(ethylene glycol)

PCL:

Poly(ε-caprolactone)

DDAB:

Dimethyldioctadecylammonium bromide

IGF-1R:

Insulin-like growth factor type I

siRNA:

Small interfering RNA

FT-IR:

Fourier transform infrared spectroscopy

1H NMR:

Proton nuclear magnetic resonance spectroscopy

AFM:

Atomic force microscopy

DLS:

Dynamic light scattering

RT-PCR:

Reverse transcription-polymerase chain reaction

PDI:

Polydispersity index

N:P ratio:

Nitrogen:Phosphate ratio

PBS:

Phosphate-buffered saline

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Acknowledgements

The authors are hereby grateful to the deputy of research and technology, and Cancer Gene Therapy Research Center, Zanjan University of Medical Sciences, Zanjan, Iran.

Funding

This study was funded by a grant (No: A-12-802-7) and ethics number (ZUMS.REC.1394.08) from Department of Clinical Biochemistry, School of Medicine, Zanjan University of Medical Sciences (ZUMS), Zanjan, Iran.

Author information

Authors and Affiliations

  1. Department of Biotechnology, School of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran

    M. Khodaei & A. H. Taromchi

  2. Department of Medicinal Chemistry, School of Pharmacy, Zanjan University of Medical Sciences, Zanjan, Iran

    K. Rostamizadeh

  3. Department of Clinical Biochemistry, School of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran

    H. Monirinasab & M. Fathi

  4. Cancer Gene Therapy Research Center, Zanjan University of Medical Sciences, Zanjan, Iran

    M. Fathi

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  1. M. Khodaei
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  2. K. Rostamizadeh
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  3. A. H. Taromchi
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  4. H. Monirinasab
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  5. M. Fathi
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Correspondence to M. Fathi.

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Ethical approval

The study was aprooved by ethical committee of Zanjan University of Medical Sciences with a number of ZUMS.REC.1394.08.

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Informed consent was obtained from all individuals participants included in the study.

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Khodaei, M., Rostamizadeh, K., Taromchi, A.H. et al. DDAB cationic lipid-mPEG, PCL copolymer hybrid nano-carrier synthesis and application for delivery of siRNA targeting IGF-1R into breast cancer cells. Clin Transl Oncol 23, 1167–1178 (2021). https://doi.org/10.1007/s12094-020-02507-3

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  • DOI: https://doi.org/10.1007/s12094-020-02507-3

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