RESEARCH ARTICLE


Comparative Activity and Specificity of Antisense Oligodeoxynucleotides and Small Interfering RNA in an in vitro Ewing Sarcoma Model



Marie Villemeur1, Ali Tamaddon1, 2, Jean-Rémi Bertrand1, Claude Malvy*, 1
1 Laboratoire de Vectorologie et Transfert de Gènes, Institut Gustave Roussy, UMR CNRS 8121, 39, rue Camille Desmoulins, 94805 Villejuif Cedex, France
2 Department of Pharmaceutics, School of Pharmacy, Shiraz University of Medical Sciences, P.O. Box 14155-6153,Tehran, Iran

Article Information


Identifiers and Pagination:

Year: 2009
Volume: 2
First Page: 36
Last Page: 45
Publisher Id: TONMJ-2-36
DOI: 10.2174/1875933500902010036

Article History:

Received Date: 29/01/2009
Revision Received Date: 19/05/2009
Acceptance Date: 28/05/2009
Electronic publication date: 26/11/2009
Collection year: 2009

© 2009 Villemeur et al.

open-access license: This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International Public License (CC-BY 4.0), a copy of which is available at: https://creativecommons.org/licenses/by/4.0/legalcode. This license permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

* Address correspondence to this author at the Laboratoire de Vectorologie et Transfert de Gènes, Institut Gustave Roussy, UMR CNRS 8121, 39, rue Camille Desmoulins, 94805 Villejuif Cedex, France; Tel: +33 142 115 402; Fax: +33 142 115 245; E-mail: cmalvy@igr.fr


Abstract

The EWS-Fli1 fusion gene, resulting from a t(11;22) translocation, plays a key role in the Ewing’s sarcoma pathogenesis. In the past, a 25mer phosphorothioate antisense oligodeoxynucleotide, a structured 30mer phosphorothioate/ phosphodiester antisense oligodeoxynucleotide, and an antisense siRNA, delivered either free, by vectors or intracellularly expressed, were found potent in various in and in Ewing sarcoma models. Because of differences among the models used in the literature, the comparison of various antisense agents with each other is difficult. Therefore, we aimed to evaluate these three antisense agents in NIH/3T3 fibroblasts which stably express the human EWS-Fli1 oncogene as an in vitro model of Ewing sarcoma. Fours parameters were considered including oncogene EWS-Fli1 and EWS mRNA expression, cellular proliferation, and actin cytoskeleton organization. They illustrate the antisense efficacy, the specificity and the phenotypic reversion for the last two ones, respectively. We showed that the structured 30mer phosphorothioate/phosphodiester antisense oligodeoxynucleotide and antisense siRNA represent the best choice for clinical trials. Nevertheless, the antisense ODN is more specific than the siRNA and represents the most efficient antisense agent. Its activity may be improved after the selection of an appropriate delivery vector which is able to increase cell penetration and to protect it from nucleases degradation.