Novel biotinylated chitosan-graft-polyethyleneimine copolymer as a targeted non-viral vector for anti-EGF receptor siRNA delivery in cancer cells.

Abstract

The major impediments to develop an efficient non-viral siRNA-mediated gene silencing method, as a therapeutic approach, are the low cellular uptake and intracellular delivery and release of non-viral vectors. To overcome these problems, designing a proper vector with high transfection efficiency is obviously under scrutiny of various studies. The present study, evaluate a novel biotinylated chitosan-graft-polyethyleneimine (Bio-Chi-g-PEI) copolymer as an appropriate non-viral vector for targeted delivery of siRNA to cancer cells. The composition of the synthesized Bio-Chi-g-PEI copolymer was thoroughly characterized using (1)H NMR and FTIR spectroscopy, besides the hydroxyazobenzene-2-carboxylic acid (HABA) assay. In vitro cytotoxicity assay of the Bio-Chi-g-PEI copolymers was performed by MTT assay. Cytotoxicity evaluations indicated that the new copolymer was markedly less toxic than PEI 25KD. Physicochemical properties of the Bio-Chi-g-PEI/siRNA complexes such as complex stability, size, zeta potential, and their morphology at various weight ratios, investigated by appropriate methods, revealed the suitability of the complexes for the transfection. The efficient cellular internalization of the complexes for HeLa and OVCAR-3 cells in culture media was confirmed by intracellular tracking of the prepared complexes using confocal laser scanning microscopy and Cy3-labeled anti-epidermal growth factor receptor siRNA. Finally, evaluation of the transfection efficiency and gene silencing by flow cytometry and real-time polymerase chain reaction highlighted the significantly higher efficiency of transfection and silencing for biotinylated copolymer compared with the PEI 25KD and non-biotinylated copolymer.

• Authors: Mohammad H Darvishi, Alireza Nomani, Mohsen Amini, Mohammad A Shokrgozar, Rassoul Dinarvand
• Journal: International journal of pharmaceutics (Int J Pharm) Vol. 456 Issue 2 Pg. 408-16 (Nov 18 2013) ISSN: 1873-3476 [Electronic] Netherlands
• PMID: 24012865 (Publication Type: Journal Article)
• Copyright: Copyright © 2013 Elsevier B.V. All rights reserved.

Chemical References

• Polymers
• RNA, Small Interfering
• chitosan-graft-polyethylenimine
• Polyethyleneimine
• Chitosan
• ErbB Receptors

Topics

• Biotinylation (methods)
• Cell Survival (drug effects, genetics)
• Chitosan (administration & dosage, analogs & derivatives, chemistry)
• Dose-Response Relationship, Drug
• ErbB Receptors (antagonists & inhibitors, genetics)
• Gene Targeting (methods)
• Gene Transfer Techniques
• Genetic Vectors (administration & dosage, chemistry)
• HeLa Cells
• Humans
• Polyethyleneimine (administration & dosage, analogs & derivatives, chemistry)
• Polymers (administration & dosage, chemistry)
• RNA, Small Interfering (administration & dosage, chemistry, genetics)