EWS/FLI1-induced manic fringe renders NIH 3T3 cells tumorigenic

May, Win; Arvand, Afsane; Thompson, Andrew; Braun, Benjamin S.; Wright, Marcienne M.; Denny, Christopher T.

doi:10.1038/ng1297-495

Cited by 141 publications

(98 citation statements)

References 23 publications

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“…To address this possibility, we investigated whether EWS/FLI1 might act to alter this pathway in an NIH3T3 model transformation system. When overexpressing EWS/FLI1, NIH3T3 demonstrate enhanced anchorage-independent growth and augmented ability to form tumors in immunodeficient mice (May et al, 1993(May et al, , 1997. Furthermore, expression of EWS/FLI1 has been shown to confer an altered cellular phenotype that to an extent mirrors that found in Ewing's tumors (Teitell et al, 1999).…”

Section: Ews/fli1 Enhances Gli1 Expression In Nih3t3 Cellsmentioning

confidence: 99%

The EWS/FLI1 oncogenic transcription factor deregulates GLI1

et al. 2007

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Ewing family tumors (EFT), classically Ewing's sarcoma and peripheral primitive neuroectodermal tumor, share a common class of tumor-specific fusion genes thought to be key mediators of tumor biology. Here we demonstrate that the most common Ewing's fusion, EWS/FLI1, produces transcriptional upregulation of GLI1 and its direct transcriptional target PATCHED1 in a model transformation system. This deregulation of GLI1 is common to other EWS/ets chimera and depends on the functional transcriptional regulatory domains. Inhibition of GLI1 via RNAi or via overexpression of endogenous inhibitors results in a reduction of EWS/FLI1 transformation activity. Activation of GLI1 appears to occur in a Hedgehog-independent fashion as blockade of Hedgehog signaling has only a modest effect on EFT cells. We present evidence that EWS/FLI1 upregulation of cMYC may play a role in the upregulation of GLI1 in EWS/ FLI1-transformed NIH3T3 cells. Finally, we demonstrate that observations made in a model transformation system translate to an Ewing cellular background. EFT cell lines express GLI1 and PATCHED and this expression is EWS/FLI1 dependent. Inhibition of GLI1 expression via RNAi results in reduced anchorage-independent growth in an EFT cell line. GLI1 appears to be a transcriptionally deregulated target of EWS/FLI1 that mediates a portion of its tumorigenic phenotype.

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Section: Ews/fli1 Enhances Gli1 Expression In Nih3t3 Cellsmentioning

confidence: 99%

The EWS/FLI1 oncogenic transcription factor deregulates GLI1

et al. 2007

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“…8 These proteins act as aberrant transcription factors leading to or initiating tumorigenic processes. 9 Fusion of EWS-FLI1 has been shown to increase expression of a number of genes, including manic fringe 10 or EWS/FLI activated transcript 2 (EAT2). 11 Furthermore, EWS fusion genes might also acquire additional nontranscriptional functions that result from the loss of RNA-binding properties of wild-type EWS.…”

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confidence: 99%

Profiling and functional annotation of mRNA gene expression in pediatric rhabdomyosarcoma and Ewing's sarcoma

Baer

Nees

Breit

et al. 2004

Intl Journal of Cancer

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Using Affymetrix oligonucleotide microarrays, we analyzed mRNA gene expression patterns of 12 primary pediatric rhabdomyosarcomas (RMS) and 11 Ewing's sarcomas (EWS), which belong to the small round blue cell tumors (SRBCTs). Diagnostic classification of these cancers is frequently complicated by the highly similar appearance in routine histology, and additional molecular markers could significantly improve tumor classification. A combination of three independent statistical approaches (t-test, SAM, k-nearest neighborhood analysis) resulted in 101 highly significant probe sets that clearly discriminate between EWS and RMS. We identified novel marker transcripts that have not been previously associated with either RMS or EWS yet, including CITED2, glypican 3 (GPC3), and cyclin D1 (CCND1). Expression levels for selected candidate genes were validated by quantitative real-time reverse-transcription PCR. Furthermore, to identify biologically meaningful trends, functional annotations were assigned to 946 genes differentially expressed between EWS and RMS (t-test). Genes involved in protein biosynthesis (n ‫؍‬ 28) and complex assembly (n ‫؍‬ 9), lipid metabolism (n ‫؍‬ 23), energy generation (n ‫؍‬ 22), and mRNA processing (n ‫؍‬ 11) were expressed significantly higher in EWS. Thus, functional annotation of tumor-specific genes reveals detailed insights into tumor biology and differentiation-specific expression patterns and gives important clues related to the possible cellular origin of these pediatric tumors. Small round blue cell tumors (SRBCTs) are frequently difficult to distinguish by standard histology or, sometimes, even by immunohistochemistry. Rhabdomyosarcoma (RMS) and Ewing's sarcomas (EWS) are common pediatric solid tumors that belong to this group that also includes neuroblastoma and Burkitt's lymphoma. However, the correct diagnosis of SRBCT tumors is essential since treatment regimens, response to therapy and prognosis may differ markedly. Therefore, molecular techniques are increasingly implemented for the diagnostic distinction of SRBCT tumors, including EWS and RMS. Progress in recent years includes the identification of characteristic tumor-specific chromosomal translocations, resulting in the expression of aberrant, oncogenic fusion proteins.RMS is the most common pediatric soft tissue sarcoma and can be histologically subdivided into the more prevalent embryonal (eRMS) and the more aggressive alveolar rhabdomyosarcoma (aRMS). ARMS have an unfavorable prognosis due to a tendency to early relapses and frequent failure of chemotherapy. This is reflected by 5-year survival rates ranging between 20 -50%. Most aRMS express PAX3-FKHR or PAX7-FKHR fusion proteins resulting from [t(2;13)(q35;q14)] or [t(1;13)(p36;q14)] translocations, respectively. 1-3 Although a frequently affected chromosomal locus in eRMS has been defined (11p15.5), specific gene(s) or a fusion transcript have not been identified. 4 Ewing's sarcomas (EWS) and the related, more differentiated peripheral neuroectodermal tumors (PNET) are...

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“…As a physiologically relevant test of transcriptional activity of EWS/FLI I347E, we evaluated endogenous expression of the EWS/FLI target gene MFNG (May et al, 1997) in our experimental system (Figure 4a). As reported previously, expression of MFNG was strongly induced in NIH3T3 cells expressing EWS/FLI (lane 3), but not in parental cells, or cells expressing wild-type FLI-1 (lanes 1 and 2).…”

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confidence: 99%

Transforming activity of EWS/FLI is not strictly dependent upon DNA-binding activity

Jaishankar

Roussel

et al. 1999

Oncogene

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In approximately 85% of Ewing sarcomas, chromosomal translocations give rise to the chimeric gene EWS/FLI, encoding the N-terminus of the RNA binding protein EWS fused to the DNA-binding domain of the ETS protein FLI-1. EWS/FLI is a stronger transcriptional activator than wild-type FLI-1, although both proteins bind to the same DNA sequences in vitro. In addition, EWS/FLI, but not FLI-1, is a transforming oncogene in NIH3T3 ®broblasts. EWS/FLI is thought to transform through its ability to deregulate the expression of target genes. We introduced several point mutations into the ETS domain of EWS/FLI that abolished DNA-binding activity. Although two of these mutations disrupted the transforming activity of EWS/FLI, one mutated protein containing a substitution of isoleucine 347 with glutamic acid (I347E) retained diminished transforming activity. In addition, EWS/FLI I347E did not activate expression of the endogenous EWS/FLI target gene manic fringe (MFNG). These studies demonstrate that a portion of the oncogenic activity of EWS/FLI is independent of FLI DNA-binding activity.

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EWS/FLI1-induced manic fringe renders NIH 3T3 cells tumorigenic

Cited by 141 publications

References 23 publications

The EWS/FLI1 oncogenic transcription factor deregulates GLI1

The EWS/FLI1 oncogenic transcription factor deregulates GLI1

Profiling and functional annotation of mRNA gene expression in pediatric rhabdomyosarcoma and Ewing's sarcoma

Transforming activity of EWS/FLI is not strictly dependent upon DNA-binding activity

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