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Targeting of EWS/FLI‐1 by RNA interference attenuates the tumor phenotype of Ewing's sarcoma cells in vitro
Author(s) -
Chansky Howard A.,
Barahmandpour Fariba,
Mei Qi,
KahnFarooqi Waqqar,
ZielinskaKwiatkowska Anna,
Blackburn Michael,
Chansky Kari,
Conrad Ernest U.,
Bruckner James D.,
Greenlee Theodore K.,
Yang Liu
Publication year - 2004
Publication title -
journal of orthopaedic research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.041
H-Index - 155
eISSN - 1554-527X
pISSN - 0736-0266
DOI - 10.1016/j.orthres.2003.12.008
Subject(s) - gene knockdown , sarcoma , cancer research , gene silencing , rna interference , ewing's sarcoma , fusion gene , small interfering rna , biology , fusion protein , cell culture , microbiology and biotechnology , rna , transfection , gene , pathology , medicine , genetics , recombinant dna
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. © 2004 Orthopaedic Research Society. Published by Elsevier Ltd. All rights reserved.