Research Article

Sterically stabilized siRNA:gold nanocomplexes enhance c-MYC silencing in a breast cancer cell model

Department of Biochemistry, Nano-Gene & Drug Delivery Laboratory, School of Life Sciences, College of Agriculture, Engineering & Science, University of KwaZulu-Natal (Westville Campus), Private Bag X54001, Durban 4000, Kwa-Zulu Natal, South Africa

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Moganavelli Singh

*Author for correspondence:

E-mail Address: singhm1@ukzn.ac.za

https://orcid.org/0000-0002-9985-6567

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Published Online:5 Jun 2019https://doi.org/10.2217/nnm-2018-0462

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Abstract

Aim: To produce sterically stabilized and functionalized gold nanoparticles (AuNPs) for efficient delivery of siRNA for c-MYC silencing in vitro. Materials & methods: Synthesized AuNPs were functionalized with chitosan and PEG₄₀₀ and PEG₂₀₀₀, morphologically and chemically characterized, and assessed for cytotoxicity and gene silencing in vitro. Results & discussion: AuNPs presented as spherical particles in the nanometer size range and successfully bound and protected the siRNA against degradation and were well tolerated in the breast adenocarcinoma (MCF-7) cell line. Nanoparticle-mediated gene knockdown studies revealed enhanced levels of c-MYC gene silencing with more than 90% reduction of MYC protein levels. Conclusion: These nanoformulations show enhanced potential for siRNA-mediated gene silencing in human breast cancer cells in vitro.

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References

1. Zhao ZX, Gao SY, Wang JC et al. Self-assembly nanomicelles based on cationic mPEG–PLA-b-Polyarginine (RI5) triblack co-polymer for siRNA delivery. Biomaterials 33(28), 6793–6807 (2010).
Google Scholar
2. Croce CM. Oncogenes and cancer. N. Engl. J. Med. 358(5), 502–511 (2008).
Medline CASGoogle Scholar
3. Ashworth A, Lord CJ, Reis-Filho JS. Genetic interactions in cancer progression and treatment. Cell 145(1), 30–38 (2011).
Medline CASGoogle Scholar
4. Dang CV, O'Donnell KA, Zeller KI, Nguyen T, Ostus RC, Li F. The c-Myc target gene network. Sem. Cancer Biol. 16(4), 253–264 (2006).
Medline CASGoogle Scholar
5. Whitehead KA, Langer R, Anderson DG. Knocking down barriers: advances in siRNA delivery. Nat. Rev. Drug Discov. 8(2), 129–138 (2009).
Medline CASGoogle Scholar
6. Sanvicens W, Marco MP. Multifunctional nanoparticles – properties and prospects for their use in human medicine. Trends Biotechnol. 26(8), 425–433 (2008).
Medline CASGoogle Scholar
7. El-Ansary A, Al-Daihan S. On the toxicity of therapeutically used nanoparticles: an overview. J. Toxicol. 2009(754180), 1–9 (2009).
Google Scholar
8. Conner EE, Mwamuka J, Gole A, Murphy CJ, Wyatt MD. Gold nanoparticles are taken up by human cells but do not cause acute toxicity. Small 1(3), 325–327 (2005). • Provides a review of the applications of gold nanoparticles (AuNPs).
MedlineGoogle Scholar
9. Ghosh P, Han G, De M, Kim CK, Rotelo VM. Gold nanoparticles in delivery applications. Adv. Drug Deliv. Rev. 60(11), 1307–1315 (2008).
Medline CASGoogle Scholar
10. Pissuwan D, Niidome T, Cortie MB. The forthcoming applications of gold nanoparticles in drug and gene delivery. J. Control. Rel. 149(1), 65–71 (2009).
MedlineGoogle Scholar
11. Cornejo-Monroy D, Acosta-Torres LS, Morena-Vega AI, Saldana C, Morales-Tlalpan V, Castaño VM. Gold nanostructures in medicine: past, present and future. J. Nanosci. Lett. 3(25), 1–9 (2013).
Google Scholar
12. Ragelle H, Vandermeulen G, Préat V. Chitosan based siRNA delivery systems. J. Control. Rel. 172(1), 207–218 (2013).
Medline CASGoogle Scholar
13. Kah JC, Wong KY, Neoh KG et al. Critical parameters in the pegylation of gold nanoshells for biomedical applications: an in vitro macrophage study. J. Drug Target. 17(3), 181–193 (2009).
Medline CASGoogle Scholar
14. Niidome T, Yamagata M, Okamoto Y et al. PEG-modified gold nanorods with a stealth character for in vivo applications. J. Control. Rel. 114(3), 343–347 (2006).
Medline CASGoogle Scholar
15. Lai WC, Liao WB. Thermo-oxidative degradation of poly (ethylene glycol)/ poly (L-lactic acid) blends. Polymer (Guildf) 44, 8103–8109 (2003).
CASGoogle Scholar
16. Vonarbourg A, Passirani C, Saulnier P, Benoit JP. Parameters influencing the stealthiness of colloidal drug delivery systems. Biomaterials 27(24), 4356–4373 (2006).
Medline CASGoogle Scholar
17. Harris JM, Martin VE, Modi M. Pegylation: a novel process for modifying pharmacokinetics. Clin. Pharmacokinet. 40(7), 539–551 (2001). •• Describes the citrate reduction synthesis of AuNPs.
Medline CASGoogle Scholar
18. Turkevitch J, Stevenson PC, Hillier J. A study of the nucleation and growth processes in the synthesis of colloidal gold. Discuss. Faraday Soc. 11, 55–75 (1951). • Provides details on the synthesis and functionalization of AuNPs.
Google Scholar
19. Lazarus GG, Revaprasadu N, López-Viota J, Singh M. The electrokinetic characterization of gold nanoparticles, functionalized with cationic functional groups, and its' interaction with DNA. Colloids Surf. B Biointerfaces 121, 425–431 (2014). •• Describes the process of polyethylene glycol(PEG)ylation of AuNPs.
Medline CASGoogle Scholar
20. Manson J, Kumar D, Meenan BJ. Polyethylene glycol functionalized gold nanoparticles: the influence of capping density on stability in various media. Gold Bull. 44(2), 99–105 (2011).
CASGoogle Scholar
21. Akinyelu J, Singh M. Chitosan stabilized gold–folate–poly(lactide-co-glycolide) nanoplexes facilitate efficient gene delivery in hepatic and breast cancer cells. J. Nanosci. Nanotechnol. 18(7), 4478–4486 (2018).
Medline CASGoogle Scholar
22. Ribble D, Goldstein N, Norris D, Shellman Y. A simple technique for quantifying apoptosis in 96-well plates. BMC Biotechnol. 5(12), 1–7 (2005).
MedlineGoogle Scholar
23. Renvoize C, Biola A, Pallardy M, Breard J. Apoptosis: identification of dying cells. Cell Biol. Toxicol. 14(2), 111–120 (1998). • Provides a description of the analysis of gene expression following real-time PCR.
Medline CASGoogle Scholar
24. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2^—ΔΔCt method. Methods 25(4), 402–408 (2001).
Medline CASGoogle Scholar
25. Thermo Scientific. ELISA Technical Guide and Protocols. Tech tip #65. Thermo Fisher Scientific Inc., MA, USA (2010).
Google Scholar
26. Wolfgang H, Nguyen TR, Thanh J, David GF. Determination of size and concentration of gold nanoparticles from UV–vis spectra. Anal. Chem. 79, 4215–4221 (2007).
MedlineGoogle Scholar
27. Nikhil RJ, Latha G, Catherine JM. Seeding growth for size control of 5–40 nm diameter gold nanoparticles. Langmuir 17, 6782–6786 (2001).
Google Scholar
28. Brown KR, Walter DG, Natan MJ. Seeding of colloidal gold nanoparticle solutions 2. Improved control of particle size and shape. Chem. Mater. 12(2), 306–313 (2000).
CASGoogle Scholar
29. Martínez JC, Chequer NA, González JL, Cardova T. Alternative methodology for gold nanoparticle diameter characterization using PCA technique and UV–vis spectrophotometry. Nanosci. Nanotechnol. 2(6), 184–189 (2012).
Google Scholar
30. Honary S, Zahir F. Effect of zeta potential on the properties of nano-drug delivery systems – a review (Part 2). Trop. J. Pharm. Res. 12(2), 265–273 (2013).
Google Scholar
31. Honary S, Zahir F. Effect of zeta potential on the properties of nano-drug delivery systems – a review (Part 1). Trop. J. Pharm. Res. 12(2), 255–264 (2013). • Provides details on the properties and applications of AuNPs.
Google Scholar
32. Mazjika A, Fülöpb L, Csapòa E et al. Functionalization of gold nanoparticles with amino acid–amyloid peptides and fragments. Colloids Surf. B Biointerfaces 81(1), 235–241 (2010).
MedlineGoogle Scholar
33. Daniel MC, Astruc D. Gold nanoparticles: assembly supramolecular chemistry, quantum-size related properties and applications toward biology, catalysis and nanotechnology. Chem. Rev. 104(1), 293–346 (2004).
Medline CASGoogle Scholar
34. Griffiths D, Bernt W, Hole P, Smith J, Mallay A, Carr B. Zeta potential measurement of nanoparticles by nanoparticle tracking analysis (NTA). NSTI-Nanotech. 1 (2011).
Google Scholar
35. Kim TH, Choi H, Yu GS, Lee J, Choi JS. Novel hyperbranched polyethyleneimine conjugate as an efficient, non-viral gene delivery vector. Macromol. Res. 21(10), 1097–1104 (2013).
CASGoogle Scholar
36. Rajam M, Pulovendran S, Rose C, Mandal AB. Chitosan nanoparticles as a dual growth factor delivery system for tissue engineering applications. Int. J. Pharm. 410(1–2), 145–152 (2011).
Medline CASGoogle Scholar
37. Boca SC, Potara M, Toderas F, Stephan O, Baldeck PL, Astilean S. Uptake and biological effects of chitosan-capped gold nanoparticles on Chinese hamster ovary cells. Mater. Sci. Eng. C 31, 184–189 (2010).
Google Scholar
38. Mbatha LS, Maiyo FC, Singh M. Dendrimer functionalized folate-targeted gold nanoparticles for luciferase gene silencing in vitro: a proof of principle study. Acta Pharm. 69(1), 49–61 (2019)
Medline CASGoogle Scholar
39. Oh KS, Kim RS, Lee J, Kim D, Cho SH, Yuk SH. Gold/chitosan/pluronic composite nanoparticles for drug delivery. J. Appl. Polymer Sci. 108(5), 3239–3244 (2008).
CASGoogle Scholar
40. Huefner A, Septiadi D, Wilts BD et al. Gold nanoparticles explore cells: cellular uptake and their use as intracellular probes. Methods 68(2), 354–363 (2014).
Medline CASGoogle Scholar
41. Huang C, Ozdemir T, Xu L-C et al. The role of substrate topography on the cellular uptake of nanoparticles. J. Biomed. Mater. Res. 104(3), 488–495 (2016).
CASGoogle Scholar

Vol. 14, No. 11

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History

Received 29 November 2018

Accepted 20 February 2019

Published online 5 June 2019

Published in print June 2019

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Financial & competing interests disclosure

The authors thank the National Research Foundation of South Africa for partial funding (M Singh, Grant ID: 113850). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

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