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Oncogenes, Fusion Genes and Tumor Suppressor Genes

Continuous Fli-1 expression plays an essential role in the proliferation and survival of F-MuLV-induced erythroleukemia and human erythroleukemia

Leukemia volume 23, pages 1311–1319 (2009)Cite this article

Abstract

Erythroleukemia induced by Friend Murine Leukemia Virus (F-MuLV) serves as a powerful tool for the study of multistage carcinogenesis and hematological malignancies in mice. Fli-1, a proto-oncogene and member of the Ets family, is activated through viral integration in F-MuLV-induced erythroleukemia, and is the most critical event in the induction of this disease. Fli-1 aberrant regulation is also observed in human malignancies, including Ewing's sarcoma, which is often linked to expression of the EWS/Fli-1 fusion oncoprotein. Here we examined the effects of Fli-1 inhibition to further elucidate its role in these pathological occurrences. The constitutive suppression of Fli-1, through RNA interference (RNAi), inhibits growth and induces death in F-MuLV-induced erythroleukemia cells. Expression of a dominant negative protein Engrailed (En)/Fli-1 reduces proliferation of EWS/Fli-1-transformed NIH-3T3 cells, and both F-MuLV-induced and human erythroleukemia cells. F-MuLV-induced erythroleukemia cells also display increased apoptosis, associated with reduced expression of bcl-2, a known fli-1 target gene. Introduction of En/Fli-1 into an F-MuLV-infected erythroblastic cell line induces differentiation, as shown by increased α-globin expression. These results suggest, for the first time, an essential role for continuous Fli-1 overexpression in the maintenance and survival of the malignant phenotype in murine and human erythroleukemias.

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References

  1. Oikawa T, Yamada T . Molecular biology of the Ets family of transcription factors. Gene 2003; 303: 11–34.

    Article  CAS  PubMed  Google Scholar 

  2. Ben-David Y, Giddens EB, Bernstein A . Identification and mapping of a common proviral integration site Fli-1 in erythroleukemia cells induced by Friend murine leukemia virus. Proc Natl Acad Sci USA 1990; 87: 1332–1336.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Klemsz MJ, Maki RAPT, Moore J, Hromas R . Characterization of the ets oncogene family member, fli-1. J Biol Chem 1994; 268: 5769–5773.

    Google Scholar 

  4. Lee CR, Cervi D, Truong AH, Li YJ, Sarkar A, Ben David Y . Friend virus-induced erythroleukemias: a unique and well-defined mouse model for the development of leukemia. Anticancer Res 2003; 23: 2159–2166.

    CAS  PubMed  Google Scholar 

  5. Tamir A, Howard J, Higgins RR, Li YJ, Berger L, Zacksenhaus E et al. Fli-1, an ets-related transcription factor, regulates erythropoietin-induced erythroid proliferation and differentiation: evidence for direct transcriptional repression of the Rb gene during differentiation. Mol Cell Biol 1999; 19: 4452–4464.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Lesault I, Quang CT, Frampton J, Ghysdael J . Direct regulation of BCL-2 by FLI-1 is involved in the survival of FLI-1-transformed erythroblasts. EMBO J 2002; 21: 694–703.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Truong AH, Cervi D, Lee J, Ben David Y . Direct transcriptional regulation of MDM2 by Fli-1. Oncogene 2005; 24: 962–969.

    Article  CAS  PubMed  Google Scholar 

  8. Pereira R, Quang CT, Lesault I, Dolznig H, Beug H, Ghysdael J . FLI-1 inhibits differentiation and induces proliferation of primary erythroblasts. Oncogene 1999; 18: 1597–1608.

    Article  CAS  PubMed  Google Scholar 

  9. Zochodne B, Truong AH, Stetler K, Higgins RR, Howard J, Dumont D et al. Epo regulates erythroid proliferation and differentiation through distinct signaling pathways: implication for erythropoiesis and Friend virus-induced erythroleukemia. Oncogene 2000; 19: 2296–2304.

    Article  CAS  PubMed  Google Scholar 

  10. Delattre O, Zucman J, Plougastel B, Desmaze C, Melot T, Peter M et al. Gene fusion with an ETS DNA-binding domain caused by chromosome translocation in human tumours. Nature 1992; 359: 162–165.

    Article  CAS  PubMed  Google Scholar 

  11. May WA, Gishizky ML, Lessnick SL, Lunsford LB, Lewis BC, Delattre O et al. Ewing sarcoma 11;22 translocation produces a chimeric transcription factor that requires the DNA-binding domain encoded by FLI1 for transformation. Proc Natl Acad Sci USA 1993; 90: 5752–5756.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Mhawech-Fauceglia P, Herrmann F, Penetrante R, Beck A, Sait S, Block AM et al. Diagnostic utility of FLI-1 monoclonal antibody and dual-color, break-apart probe fluorescence in situ (FISH) analysis in Ewing's sarcoma/primitive neuroectodermal tumor (EWS/PNET). A comparative study with CD99 and FLI-1 polyclonal antibodies. Histopathology 2006; 49: 569–575.

    Article  CAS  PubMed  Google Scholar 

  13. Cui JW, Li YJ, Sarkar A, Brown J, Tan YH, Premyslova M et al. Retroviral insertional activation of the Fli-3 locus in erythroleukemias encoding a cluster of microRNAs that convert Epo-induced differentiation to proliferation. Blood 2007; 110: 2631–2640.

    Article  CAS  PubMed  Google Scholar 

  14. Funk WD, Wright WE . Cyclic amplification and selection of targets for multicomponent complexes: myogenin interacts with factors recognizing binding sites for basic helix-loop-helix, nuclear factor 1, myocyte-specific enhancer-binding factor 2, and COMP1 factor. Proc Natl Acad Sci USA 1992; 89: 9484–9488.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Howard JC, Berger L, Bani MR, Hawley R, Ben-David Y . Activation of the erythropoietin gene in the majority of F-MuLV-induced erythroleukemias results in growth factor independence and enhanced tumorigenicity. Oncogene 1996; 12: 1405–1415.

    CAS  PubMed  Google Scholar 

  16. Wang N, Kudryavtseva E, Ch'en IL, McCormick J, Sugihara TM, Ruiz R et al. Expression of an engrailed-LMO4 fusion protein in mammary epithelial cells inhibits mammary gland development in mice. Oncogene 2004; 23: 1507–1513.

    Article  CAS  PubMed  Google Scholar 

  17. Markel H, Chandler J, Werr W . Translational fusions with the engrailed repressor domain efficiently convert plant transcription factors into dominant-negative functions. Nucleic Acids Res 2002; 30: 4709–4719.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Montross WT, Ji H, McCrea PD . A beta-catenin/engrailed chimera selectively suppresses Wnt signaling. J Cell Sci 2000; 113: 1759–1770.

    CAS  PubMed  Google Scholar 

  19. Lyon JJ, Watson RJ . Interference of Myb transactivation activity by a conditional dominant negative protein: functional interference in a cytotoxic T-cell line results in G1 arrest. Gene 1996; 182: 123–128.

    Article  CAS  PubMed  Google Scholar 

  20. Watson DK, Smyth FE, Thompson DM, Cheng JQ, Testa JR, Papas TS et al. The ERGB/fli-1 gene: isolation and characterization of a new member of the family of human ETS transcription factors. Cell Growth Differ 1992; 3: 705–713.

    CAS  PubMed  Google Scholar 

  21. Rao VN, Ohno T, Prasad DD, Bhattacharya G, Reddy ES . Analysis of the DNA-binding and transcriptional activation functions of human Fli-1 protein. Oncogene 1993; 8: 2167–2173.

    CAS  PubMed  Google Scholar 

  22. Zhang L, Lemarchandel V, Romeo P-H, Ben-David Y, Bernstein A . The Fli-1 proto-oncogene involved in erythroleukemia and Ewing's sarcoma, encodes a transcriptional activator with DNA-binding specificities distinct from other Ets family members. Oncogene 1993; 8: 1621–1630.

    CAS  PubMed  Google Scholar 

  23. Ben-David Y, Giddens EG, Letwin K, Bernstein A . Erythroleukemia induction by Friend murine leukemia virus: insertional activation of a new member of the ets gene family, Fli-1, closely linked to c-ets-1. Genes Dev 1991; 5: 908–918.

    Article  CAS  PubMed  Google Scholar 

  24. Howard JC, Li Q, Chu W, Zochodne B, Kapoor M, Ung Y et al. Bcl-2 expression in F-MuLV-induced erythroleukemias: a role for the anti-apoptotic action of Bcl-2 during tumor progression. Oncogene 2001; 20: 2291–2300.

    Article  CAS  PubMed  Google Scholar 

  25. Ben-David Y, Bernstein A . Friend virus-induced erythroleukemia and the multistage nature of cancer. Cell 1991; 66: 831–834.

    Article  CAS  PubMed  Google Scholar 

  26. Zhang L, Eddy A, Teng T, Fritzler M, Kluppel M, Melet F et al. An immunological renal disease in transgenic mice that overexpress Fli-1, a member of the ets family of transcription. Mol Cell Biol 1995; 15: 6961–6970.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Hart A, Melet F, Grossfeld P, Chien K, Jones C, Tunnacliffe A et al. Fli-1 is required for murine vascular and megakaryocytic development and is hemizygously deleted in patients with thrombocytopenia. Immunity 2000; 13: 167–177.

    Article  CAS  PubMed  Google Scholar 

  28. Spyropoulos DD, Pharr PN, Lavenburg KR, Jackers P, Papas TS, Ogawa M et al. Hemorrhage, impaired hematopoiesis, and lethality in mouse embryos carrying a targeted disruption of the Fli1 transcription factor. Mol Cell Biol 2000; 20: 5643–5652.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Melet F, Motro B, Rossi DJ, Zhang L, Bernstein A . Generation of a novel Fli-1 protein by gene targeting leads to a defect in thymus development and a delay in friend virus-induced erythroleukemia. Mol Cell Biol 1996; 16: 2708–2715.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. May WA, Lessnick SL, Braun BS, Klemsz M, Lewis BC, Lunsford LB et al. The Ewing's sarcoma EWS/FLI-1 fusion gene encodes a more potent transcriptional activator and is a more powerful transforming gene than FLI-1. Mol Cell Biol 1993; 13: 7393–7398.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. May WA, Arvand A, Thompson AD, Braun BS, Wright M, Denny CT . EWS/FLI1-induced manic fringe renders NIH 3T3 cells tumorigenic. Nat Genet 1997; 17: 495–497.

    Article  CAS  PubMed  Google Scholar 

  32. Dohjima T, Lee NS, Li H, Ohno T, Rossi JJ . Small interfering RNAs expressed from a Pol III promoter suppress the EWS/Fli-1 transcript in an Ewing sarcoma cell line. Mol Ther 2003; 7: 811–816.

    Article  CAS  PubMed  Google Scholar 

  33. Torchia EC, Boyd K, Rehg JE, Qu C, Baker SJ . EWS/FLI-1 induces rapid onset of myeloid/erythroid leukemia in mice. Mol Cell Biol 2007; 27: 7918–7934.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Yi H, Fujimura Y, Ouchida M, Prasad DD, Rao VN, Reddy ES . Inhibition of apoptosis by normal and aberrant Fli-1 and erg proteins involved in human solid tumors and leukemias. Oncogene 1997; 14: 1259–1268.

    Article  CAS  PubMed  Google Scholar 

  35. Goltzene F, Skalski M, Wolff CM, Meyer D, Mager-Heckel AM, Darribere T et al. Heterotopic expression of the Xl-Fli transcription factor during Xenopus embryogenesis: modification of cell adhesion and engagement in the apoptotic pathway. Exp Cell Res 2000; 260: 233–247.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

We would like to thank Dr C Danny for providing us with the EWS-Fli-1 construct, Dr D Barber for the VSVG and gag/pol expression vectors, and Dr MI Arnone for the En/Erg expression vector. This work was supported by a grant from the Terry Fox Foundation through the National Cancer Institute of Canada (NCIC) and the Canadian Institute of Health Research (CIHR) to YBD.

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Author notes

  1. J-W Cui and L M Vecchiarelli-Federico: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Molecular and Cellular Biology, Sunnybrook Health Sciences Centre, Toronto, Canada

    J-W Cui, L M Vecchiarelli-Federico, Y-J Li & Y Ben-David

  2. Department of Hematology, First Hospital of Jilin University, Changchun, China

    J-W Cui & G-J Wang

  3. Department of Medical Biophysics, University of Toronto, Toronto, Canada

    L M Vecchiarelli-Federico & Y Ben-David

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  1. J-W Cui
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Correspondence to Y Ben-David.

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Supplementary Information accompanies the paper on the Leukemia website (http://www.nature.com/leu)

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Cui, JW., Vecchiarelli-Federico, L., Li, YJ. et al. Continuous Fli-1 expression plays an essential role in the proliferation and survival of F-MuLV-induced erythroleukemia and human erythroleukemia. Leukemia 23, 1311–1319 (2009). https://doi.org/10.1038/leu.2009.20

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