Nanolipoparticles-mediated MDR1 siRNA delivery: preparation, characterization and cellular uptake

Document Type : Research Paper

Authors

1 Biotechnology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran

2 Nanotechnology Research Center, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

3 Charite´ Campus Mitte, Institute of Pathology, Charite´platz 1, D-10117 Berlin, Germany

4 Pharmaceutical Research Center, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

5 Department of Pharmaceutical Biotechnology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

Abstract

Objective(s):
Lipid-based nanoparticles (NLP) are PEGylated carriers composed of lipids and encapsulated nucleic acids with a diameter less than 100 nm. The presence of PEG in the NLP formulation improves the particle pharmacokinetic behavior. The purpose of this study was to prepare and characterize NLPs containing MDR1 siRNA and evaluate their cytotoxicity and cellular uptake. MDR1 siRNA could be used in multidrug resistance reversal in cancer therapy.
Materials and Methods:
siRNAs were encapsulated into NLPs consisted of mPEG-DSPE/DOTAP/DOPE (10:50:40 molar ratio) by the detergent dialysis method. The particle diameters of NLPs and their surface charge were measured using dynamic light scattering. siRNA encapsulation efficiency was determined by an indirect method via filtration and free siRNA concentration determination. NLPs cytotoxicity was investigated by MTT assay. The ability of NLPs for siRNA delivery checked in two human cell lines (MCF-7/ADR and EPP85-181/RDB) by fluorescence microscopy and compared with oligofectamine.
Results:
NLPs containing MDR1 siRNA were prepared with the stable size of 80-90 nm and the zeta potential near to neutral. The siRNA encapsulation efficacy was more than 80%. These properties are suitable for in vivo siRNA delivery. NLPs cytotoxicity studies demonstrated they were non-toxic at the doses used. NLPs improved siRNA localization in both cell lines.
Conclusion:
NLPs containing MDR1 siRNA can be a good candidate for in vivo siRNA delivery studies.

Keywords

References

  1. Abbasi M, Lavasanifar A, Uludaˇ H. Recent attempts at RNAi‐mediated P‐glycoprotein downregulation for reversal of multidrug resistance in cancer. Med Res Rev. 2013; 33(1): 33-53.
  2. Li W, Szoka FC, Jr. Lipid-based nanoparticles for nucleic acid delivery. Pharm Res. 2007; 24(3): 438-449.
  3. Seow Y, Wood MJ. Biological gene delivery vehicles: beyond viral vectors. Mol Ther. 2009; 17(5): 767-777.
  4. Mo RH, Zaro JL, Ou J-HJ, Shen W-C. Effects of Lipofectamine 2000/siRNA complexes on autophagy in hepatoma cells. Mol Biotechnol. 2012; 51(1): 1-8.
  5. Stierle V, Laigle A, Jolles B. The reduction of P-glycoprotein expression by small interfering RNAs is improved in exponentially growing cells. Oligonucleotides. 2004; 14(3): 191-198.
  6. Huang L, Liu Y. In vivo delivery of RNAi with lipid-based nanoparticles. Annu Rev Biomed Eng. 2011; 13: 507-530.
  7. Fenske DB, Cullis PR. Liposomal nanomedicines. Expert Opin Drug Deliv. 2008; 5(1): 25-44.
  8. Wheeler JJ, Palmer L, Ossanlou M, MacLachlan I, Graham RW, Zhang YP, et al. Stabilized plasmid-lipid particles: construction and characterization. Gene Ther. 1999; 6(2): 271-281.
  9. Monck MA, Mori A, Lee D, Tam P, Wheeler JJ, Cullis PR, et al. Stabilized plasmid-lipid particles: pharmacokinetics and plasmid delivery to distal tumors following intravenous injection. J Drug Target. 2000; 7(6): 439-452.
  10. Ambegia E, Ansell S, Cullis P, Heyes J, Palmer L, MacLachlan I. Stabilized plasmid-lipid particles containing PEG-diacylglycerols exhibit extended circulation lifetimes and tumor selective gene expression. Biochim Biophys Acta. 2005; 1669(2): 155-163.
  11. Li W, Huang Z, MacKay JA, Grube S, Szoka FC, Jr. Low-pH-sensitive poly(ethylene glycol) (PEG)-stabilized plasmid nanolipoparticles: effects of PEG chain length, lipid composition and assembly conditions on gene delivery. J Gene Med. 2005; 7(1): 67-79.
  12. Morandat S, El Kirat K. Solubilization of supported lipid membranes by octyl glucoside observed by time-lapse atomic force microscopy. Colloids Surf B Biointerfaces. 2007; 55(2): 179-184.
  13. Iman M, Huang Z, Szoka FC, Jr., Jaafari MR. Characterization of the colloidal properties, in vitro antifungal activity, antileishmanial activity and toxicity in mice of a di-stigma-steryl-hemi-succinoyl-glycero-phosphocholine liposome-intercalated amphotericin B. Int J Pharm. 2011; 408(1-2): 163-172.
  14. Kowalski P, Surowiak P, Lage H. Reversal of different drug-resistant phenotypes by an autocatalytic multitarget multiribozyme directed against the transcripts of the ABC transporters MDR1/P-gp, MRP2, and BCRP. Mol Ther. 2005; 11(4): 508-522.
  15. Ebrahimnejad P, Dinarvand R, Sajadi A, Jaafari MR, Nomani AR, Azizi E, et al. Preparation and in vitro evaluation of actively targetable nanoparticles for SN-38 delivery against HT-29 cell lines. Nanomed Nanotech Biol Med. 2010; 6(3): 478-485.
  16. WI M. Label IT® siRNA Tracker Intracellular Localization Kit- Mirus Bio LLC. ww.mirusbio.com/assets/protocols/ml035_label_it_sirna_tracker_kit.pdf10.4.2012. 
Nanomedicine Journal
Volume 2, Issue 1 - Serial Number 1
January 2015
Pages 39-45

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