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EWS/FLI-1 oncoprotein subtypes impose different requirements for transformation and metastatic activity in a murine model

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Abstract

Ewing sarcoma/primitive neuroectodermal tumors (EWS/PNET) are characterized by specific chromosomal translocations most often generating a chimeric EWS/FLI-1 gene. Depending on the number of juxtaposed exons assembled, several fusion types have been described with different incidences and prognoses. To assess the impact of each fusion type on the specific phenotypic, tumorigenic, and metastatic features of EWS/PNET, we developed an amenable system using a murine mesenchymal multipotent C3H10T1/2 cell line. Upon transduction of EWS/FLI-1, cells acquired dramatic morphological changes in vitro, including a smaller size and “neurite-like” membrane elongations. Chimeric fusion proteins conferred oncogenic properties in vitro, including anchorage-independent growth and an increased rate of proliferation. Furthermore, EWS/FLI-1 expression blocked mineralization, with concomitant repression of osteoblastic genes, and induced a dramatic repression of the adipocytic differentiation program. Moreover, EWS/FLI-1 promoted an aberrant neural phenotype by the de novo expression of specific neural genes. The intramuscular injection of transduced cells led to tumor development and the induction of overt osteolytic lesions. Analogously, to what was observed in human tumors, type 2 EWS/FLI-1 cells formed primary tumors in immunodeficient mice with a higher incidence and a lower latency than cells bearing types 1 and 3 fusions. By contrast, cells expressing types 2 and 3 fusions showed specific metastatic activity with a higher number of macroscopic metastases in soft tissues and osteolytic lesions in the limbs as compared to type-1-expressing cells. Therefore, the structure of each oncoprotein strongly influenced its tumorigenicity and metastagenicity. Thus, this model provides a basis for understanding the genetic determinants involved in Ewing tumor development and metastatic activity and represents a cellular system to analyze other oncoproteins involved in human sarcomagenesis.

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Acknowledgment

We thank Dr. L. Montuenga, Dr. J. A. Martínez-Climent, and Josune Orbe for the useful discussions. We are grateful to Dr. M. San Julián, Dr. L. Sierrasesúmaga, Dr. D. Lozano, Dr. E. Bandrés and especially Dr. A. Patiño for the valuable insights and comments. We also thank S. Martínez and C. Zandueta for the excellent technical assistance, the members of the Morphology Core facility, especially A. Urbiola, D. García-Ros, J. Guillén, and all the members of CIMA’s Animal Core Facility. We are indebted to Prof. R. Jordana for his valuable help in the scanning microscopy. I.G. is a Postdoctoral Fellow of the Government of Navarra and the Foundation for Applied Medical Research (FIMA). E.A. is an Associate Research Professor of the CSIC (National Research Council). F.L. is an investigator from “Ramón y Cajal” Program. This work was supported by “UTE project FIMA” agreement and the Spanish Ministry of Health/Fondo de Investigaciones Sanitarias-Feder (RTICCC C03/10), PI020828 (to EA), ISC-RETIC RD06/0020, and PI042284 (to FL). F.L. is also supported by funds from the “La Caixa Foundation” and is a recipient of the “Ortiz de Landázuri” award (67/2005, Government of Navarra).

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Authors and Affiliations

  1. Molecular Pathology Program, Centro de Investigación del Cáncer/IBMCC, University of Salamanca/CSIC, Salamanca, Spain

    Enrique de Alava

  2. Division of Oncology, Adhesion and Metastasis Laboratory, Center for Applied Medical Research (CIMA), University of Navarra, Pamplona, 31080, Navarra, Spain

    Iranzu González, Silvestre Vicent & Fernando Lecanda

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  1. Iranzu González
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  2. Silvestre Vicent
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  3. Enrique de Alava
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  4. Fernando Lecanda
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Correspondence to Fernando Lecanda.

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González, I., Vicent, S., de Alava, E. et al. EWS/FLI-1 oncoprotein subtypes impose different requirements for transformation and metastatic activity in a murine model. J Mol Med 85, 1015–1029 (2007). https://doi.org/10.1007/s00109-007-0202-5

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  • DOI: https://doi.org/10.1007/s00109-007-0202-5

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