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

机译:在体外尤因肉瘤模型中反义寡核苷酸和小干扰RNA的比较活性和特异性

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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 vitro and in vivo 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.
机译:由t(11; 22)易位产生的EWS-Fli1融合基因在尤因肉瘤的发病机理中起着关键作用。过去,人们发现在各种体外和体内尤因肉瘤模型中,通过载体自由表达或在细胞内表达的25mer硫代磷酸酯反义寡聚脱氧核苷酸,结构化的30mer硫代磷酸酯/磷酸二酯反义寡聚核苷酸和反义siRNA是有效的。由于文献中使用的模型之间存在差异,因此很难将各种反义剂彼此进行比较。因此,我们旨在评估NIH / 3T3成纤维细胞中的这三种反义剂,该成纤维细胞稳定表达人EWS-Fli1癌基因作为尤因肉瘤的体外模型。考虑了四个参数,包括致癌基因EWS-Fli1和EWS mRNA表达,细胞增殖和肌动蛋白细胞骨架组织。它们分别说明了后两个的反义功效,特异性和表型逆转。我们表明,结构化的30mer硫代磷酸酯/磷酸二酯反义寡聚脱氧核苷酸和反义siRNA代表了临床试验的最佳选择。然而,反义ODN比siRNA更具特异性,是最有效的反义剂。在选择能够增加细胞渗透并保护其免受核酸酶降解的合适的递送载体之后,可以改善其活性。

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