Stable interference of EWS-FLI1 in an Ewing sarcoma cell line impairs IGF-1/IGF-1R signalling and reveals TOPK as a new target

被引:86
作者
Herrero-Martin, D. [1 ]
Osuna, D. [1 ]
Ordonez, J. L. [1 ]
Sevillano, V. [1 ]
Martins, A. S. [1 ]
Mackintosh, C. [1 ]
Campos, M. [1 ]
Madoz-Gurpide, J. [1 ]
Otero-Motta, A. P. [1 ]
Caballero, G. [1 ]
Amaral, A. T. [1 ]
Wai, D. H. [2 ]
Braun, Y. [3 ]
Eisenacher, M. [4 ]
Schaefer, K-L [3 ]
Poremba, C. [3 ]
de Alava, E. [1 ]
机构
[1] USAL CSIC, Ctr Invest Canc IBMCC, Mol Pathol Program, Salamanca 37007, Spain
[2] Childrens Hosp Los Angeles, Dept Pathol, Los Angeles, CA 90027 USA
[3] Univ Dusseldorf, Inst Pathol, D-40225 Dusseldorf, Germany
[4] Univ Munster, Dept Integrated Funct Gen, D-48149 Munster, Germany
关键词
Ewing sarcoma; EWS-FLI1; stable shRNAi model; IGF-1/IGF-1R; TOPK; ORIGINATED PROTEIN-KINASE; PDZ-BINDING KINASE; MITOTIC KINASE; EXPRESSION; EWS/FLI; GROWTH; TUMOR; GENE; PHOSPHORYLATION; TRANSFORMATION;
D O I
10.1038/sj.bjc.6605104
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
BACKGROUND: Ewing sarcoma is a paradigm of solid tumour-bearing chromosomal translocations resulting in fusion proteins that act as deregulated transcription factors. Ewing sarcoma translocations fuse the EWS gene with an ETS transcription factor, mainly FLI1. Most of the EWS-FLI1 target genes still remain unknown and many have been identified in heterologous model systems. METHODS: We have developed a stable RNA interference model knocking down EWS-FLI1 in the Ewing sarcoma cell line TC71. Gene expression analyses were performed to study the effect of RNA interference on the genetic signature of EWS-FLI1 and to identify genes that could contribute to tumourigenesis. RESULTS: EWS-FLI1 inhibition induced apoptosis, reduced cell migratory and tumourigenic capacities, and caused reduction in tumour growth. IGF-1 was downregulated and the IGF-1/IGF-1R signalling pathway was impaired. PBK/TOPK (T-LAK cell-originated protein kinase) expression was decreased because of EWS-FLI1 inhibition. We showed that TOPK is a new target gene of EWS-FLI1. TOPK inhibition prompted a decrease in the proliferation rate and a dramatic change in the cell's ability to grow in coalescence. CONCLUSION: This is the first report of TOPK activity in Ewing sarcoma and suggests a significant role of this MAPKK-like protein kinase in the Ewing sarcoma biology. British Journal of Cancer ( 2009) 101, 80-90. doi: 10.1038/sj.bjc.6605104 www.bjcancer.com Published online 2 June 2009 (C) 2009 Cancer Research UK
引用
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页码:80 / 90
页数:11
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