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Anti-epidermal growth factor receptor siRNA carried by chitosan-transacylated lipid nanocapsules increases sensitivity of glioblastoma cells to temozolomide

Authors Messaoudi K, Saulnier P, Boesen K, Benoit JP, Lagarce F

Received 13 December 2013

Accepted for publication 22 January 2014

Published 24 March 2014 Volume 2014:9(1) Pages 1479—1490

DOI https://doi.org/10.2147/IJN.S59134

Checked for plagiarism Yes

Review by Single-blind

Peer reviewer comments 5

Khaled Messaoudi,1 Patrick Saulnier,1 Kim Boesen,1 Jean-Pierre Benoit,1,2 Frederic Lagarce1,2

1L'Université Nantes Angers Le Mans, INSERM U1066, Micro et nanomédecines biomimétiques, Angers, France; 2Pharmacy Department, Angers University Hospital, Angers, France

Abstract: Epidermal growth factor receptor (EGFR) is a crucial protein that plays an important role in the maintenance and development of glioblastomas. The silencing or knockdown of EGFR is possible by administering a small interfering ribonucleic acid (siRNA). Lipid nanocapsules (LNCs) covered by chitosan were developed in our laboratory by a transacylation process. The resulting nanocapsules have a positive zeta potential that enables electrostatic interactions with the negatively-charged siRNA. Prior to transfection, the cytotoxicity of the nanocapsules by (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) (MTS) test was performed on the U87MG cell line to determine non-toxic levels of the LNCs to avoid cell mortality. Treatment of the U87MG cells with the chitosan-transacylated LNCs/anti-EGFR siRNA complex resulted in a reduction of EGFR expression by 51.95%±6.03% (P≤0.05) after 96 hours of incubation. It also increased the cellular sensitivity to temozolomide in comparison to untreated cells with siRNA. The largest increase in mortality was 62.55%±3.55% (P<0.05). This successful knockdown provides proof for the concept of surface grafting of siRNA onto LNCs to modify cell sensitivity to temozolomide. The method could be implemented in future clinical models regarding the experimental treatment of glioblastoma cancer.

Keywords: EGFR, glioblastoma, siRNA, lipid nanocapsules, chitosan, temozolomide

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