Analysis of the expression of cell cycle regulators in Ewing cell lines: EWS-FLI-1 modulates p57KIP2and c-Myc expression

Oncogene. 2001 May 31;20(25):3258-65. doi: 10.1038/sj.onc.1204437.

Abstract

Ewing tumour is characterized by specific chromosome translocations which fuse EWS to a subset of genes encoding ETS transcription factors, most frequently FLI-1. We report the analysis of the expression of various cell cycle regulators both in Ewing tumour derived cell lines and in different cellular models with either inducible or constitutive EWS-FLI-1 cDNA expression. In Ewing cell lines, cyclin D1, CDK4, Rb, p27KIP1 and c-Myc were consistently highly expressed whereas p57KIP2, p15INK4B and p14ARF demonstrated undetectable or low expression levels. The amount of p16INK4A, p21CIP1, p18INKAC and CDK6 was variable from one cell line to the other. The inducible expression of EWS-FLI-1 led to a strong upregulation of c-Myc and a considerable downregulation of p57KIP2. Other proteins did not show evident modification. High c-Myc and very low p57KIP2 expression levels were also observed in neuroblastoma NGP cells constitutively expressing EWS-FLI-1 as compared to parental cells. Analysis of the p57KIP2 promoter indicated that EWS-FLI-1 downregulates, possibly through an indirect mechanism, the transcription of this gene. Finally, we show that ectopic expression of p57KIP2 in Ewing cells blocks proliferation through a complete G1 arrest. These results suggest that the modulation of p57(KIP2) expression by EWS-FLI-1 is a fundamental step in Ewing tumorigenesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Cell Cycle
  • Cell Cycle Proteins / metabolism*
  • Cell Transformation, Neoplastic
  • Cyclin-Dependent Kinase Inhibitor p57
  • Down-Regulation
  • Humans
  • Molecular Sequence Data
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism*
  • Oncogene Proteins, Fusion / metabolism*
  • Promoter Regions, Genetic
  • Proto-Oncogene Protein c-fli-1
  • Proto-Oncogene Proteins c-myc / metabolism*
  • RNA-Binding Protein EWS
  • Sarcoma, Ewing / etiology*
  • Transcription Factors / metabolism*
  • Tumor Cells, Cultured

Substances

  • CDKN1C protein, human
  • Cell Cycle Proteins
  • Cyclin-Dependent Kinase Inhibitor p57
  • EWS-FLI fusion protein
  • Nuclear Proteins
  • Oncogene Proteins, Fusion
  • Proto-Oncogene Protein c-fli-1
  • Proto-Oncogene Proteins c-myc
  • RNA-Binding Protein EWS
  • Transcription Factors