Targeting of EWS/FLI-1 by RNA interference attenuates the tumor phenotype of Ewing's sarcoma cells in vitro
Targeting of EWS/FLI-1 by RNA interference attenuates the tumor phenotype of Ewing's sarcoma cells in vitro
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DOI：
10.1016/j.orthres.2003.12.008
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发表时间：
2004-07-01
影响因子：
2.8
通讯作者：
Yang, L
中科院分区：
文献类型：
--
作者：
Chansky, HA;Barahmand-pour, F;Yang, L
尤文氏肉瘤的细胞遗传学异常表现为表达EWS/FLI-1嵌合融合蛋白的平衡t（11 -.22）易位。EWS/FLI-1的作用似乎是显性负性的，因为EWS的过表达不能克服肉瘤表型。先前的研究表明，EWS/FLI-1以及相关的肉瘤融合蛋白是必要的，足以在体外和体内诱导转化。在这项研究中，我们报告，合成的小干扰RNA（siRNA）特异性抑制EWS/FLI-1融合基因在SK-ES尤文肉瘤细胞的表达。EWS/FLI-1融合蛋白的敲低与细胞增殖降低和细胞凋亡增加相关。我们证明尤文肉瘤肿瘤以及尤文肉瘤细胞系主要表达CXCR 4趋化因子受体。使用体外侵袭测定，CXCR 4的SDF-1配体显示为SK-ES细胞侵袭的有效刺激物。通过RNA干扰敲低EWS/FLI-1可消除SK-ES细胞的侵袭性。这些实验表明，靶向沉默EWS/FLI-1融合基因的siRNA代表了一个有前途的策略，研究EWS/FLI-1蛋白在尤文肉瘤细胞的损失，否则相同的遗传背景。(C)2004骨科研究学会。由爱思唯尔有限公司出版。保留所有权利。
The defining cytogenetic abnormality of Ewing's sarcoma is the presence of a balanced t(11 -.22) translocation expressing the EWS/FLI-1 chimeric fusion protein. The effect of EWS/FLI-1 appears to be dominant negative since over-expression of EWS does not overcome the sarcoma phenotype. Previous studies have shown that EWS/FLI-1 as well as related sarcoma fusion proteins are necessary and sufficient to induce transformation both in vitro and in vivo. In this study we report that synthetic small interfering RNA (siRNA) specifically suppresses EWS/FLI-1 fusion gene expression in SK-ES Ewing's sarcoma cells. Knockdown of the EWS/FLI-1 fusion protein is correlated with decreased cell proliferation and increased apoptosis. We demonstrate that Ewing's sarcoma tumors as well as Ewing's sarcoma cell lines predominantly express the CXCR4 chemokine receptor. Using an in vitro invasion assay, the SDF-1 ligand of CXCR4 was shown to be a potent stimulus of invasion by SK-ES cells. Knockdown of EWS/FLI-1 by RNA interference abrogates the invasiveness of SK-ES cells. These experiments suggest that targeted silencing of the EWS/FLI-1 fusion gene by siRNA represents a promising strategy to study the loss of EWS/FLI-1 protein in Ewing's sarcoma cells of otherwise identical genetic background. (C) 2004 Orthopaedic Research Society. Published by Elsevier Ltd. All rights reserved.