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Pharmaceutical Research

, Volume 32, Issue 12, pp 3813–3826 | Cite as

Effect of siRNA pre-Exposure on Subsequent Response to siRNA Therapy

  • Hamidreza Montazeri AliabadiEmail author
  • Parvin Mahdipoor
  • Cezary Kucharsky
  • Nicole Chan
  • Hasan UludağEmail author
Research Paper

ABSTRACT

Purpose

An alternative cancer therapy based on RNA interference (RNAi) has shown considerable promise but the possibility of resistance development is not known. This study explored the possibility of therapeutic resistance against siRNA nanoparticles in human cancer cells.

Methods

Two approaches to siRNA treatment were undertaken using lipid-modified polyethylenimines, a single high concentration (shock) and repeated increasing concentrations (gradual). The targets were Mcl-1, RPS6KA5 and KSP in MDA-MB-435 cells.

Results

There was no evidence of resistance development in shock-treated cells, while the decrease in mRNA levels of targeted proteins was not as robust in naïve cells in gradual treatment. However, silencing efficiency was restored after a 7-day recovery period when expression of suppressed proteins returned to normal levels. Cellular uptake of siRNA was not affected by pre-treatments. Other mediators involved in cell survival and proliferation were altered in siRNA-treated cells, but only JUN silencing led to a heightened loss of viability. In vivo experiments demonstrated similar silencing efficiency at mRNA level after repeat doses.

Conclusions

Human cancer cells responded to repeat siRNA nanoparticles in a similar fashion after a temporary initial alteration and little, if any, resistance was evident against repeated siRNA treatments.

KEY WORDS

siRNA cancer therapy resistance polymeric carriers 

ABBREVIATIONS

FAM

Fluorescein amidite

KSP

Kinesin spindle protein

Mcl-1

Myeloid leukemia cell differentiation protein

mRNA

Messenger RNA

NT

Non-treated

PEI-LA

Linoleic acid-substituted polyethylenimine

RISC

RNA-induced silencing complex

RQ

Relative quantity

RT-PCR

Real-time polymerase chain reaction

siRNA

Short interfering RNA

Notes

ACKNOWLEDGMENTS AND DISCLOSURES

The authors would like to thank Mrs. Geraldine Barron for her invaluable help with development and optimization of confocal microscopy, and Dr. Robert Clarke (Georgetown University, Washington, DC) for providing the MDA-435 cells. This project was financially supported by a Breast Cancer Research Grant from Canadian Breast Cancer Foundation (CBCF) and a NSERC Discovery Grant.

Supplementary material

11095_2015_1741_MOESM1_ESM.pptx (62 kb)
Supplementary Figure 1 (PPTX 62 kb)

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Hamidreza Montazeri Aliabadi
    • 1
    • 2
    Email author
  • Parvin Mahdipoor
    • 2
  • Cezary Kucharsky
    • 2
  • Nicole Chan
    • 2
  • Hasan Uludağ
    • 2
    • 3
    • 4
    Email author
  1. 1.School of PharmacyChapman UniversityIrvineUSA
  2. 2.Department of Chemical & Material Engineering, Faculty of EngineeringUniversity of AlbertaEdmontonCanada
  3. 3.Faculty of Pharmacy and Pharmaceutical SciencesUniversity of AlbertaEdmontonCanada
  4. 4.Department of Biomedical Engineering, Faculty of Medicine and DentistryUniversity of AlbertaEdmontonCanada

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