A monoclonal antibody specifically reactive with Ewing's sarcoma

Hara, Shinichi; Ishii, Eizaburo; Tanaka, Shigeki; Yokoyama, Jotaro; Katsumata, Kiyomitsu; Fujimoto, Junichiro; Hata, Junichi

doi:10.1038/bjc.1989.383

Cited by 26 publications

(9 citation statements)

References 7 publications

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“…UET-13 cells were cultured in Dulbecco's modified Eagle's medium (DMEM) with 10% Tet system approved fetal bovine serum (T-FBS) (Takara) at 37°C under a humidified 5% CO 2 atmosphere. EFT cell lines (EES-1 [20], SCCH196 [21], RD-ES and SK-ES1 [5], NCR-EW2 and NCR-EW3 [19], and W-ES [13]) and neuroblastoma (NB) cell lines (NB69 and NB9 [15] and GOTO [47]) were cultured in RPMI 1640 with 10% FBS. A rhabdomyosarcoma cell line, NRS-1 (40), was cultured in Eagle's minimal essential medium with 10% FBS.…”

Section: Methodsmentioning

confidence: 99%

Inducible Expression of Chimeric EWS/ETS Proteins Confers Ewing's Family Tumor-Like Phenotypes to Human Mesenchymal Progenitor Cells

Miyagawa

Okita

Nakaijima

et al. 2008

Molecular and Cellular Biology

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Ewing's family tumor (EFT) is a rare pediatric tumor of unclear origin that occurs in bone and soft tissue. Specific chromosomal translocations found in EFT cause EWS to fuse to a subset of ets transcription factor genes (ETS), generating chimeric EWS/ETS proteins. These proteins are believed to play a crucial role in the onset and progression of EFT. However, the mechanisms responsible for the EWS/ETS-mediated onset remain unclear. Here we report the establishment of a tetracycline-controlled EWS/ETS-inducible system in human bone marrow-derived mesenchymal progenitor cells (MPCs). Ectopic expression of both EWS/FLI1 and EWS/ERG proteins resulted in a dramatic change of morphology, i.e., from a mesenchymal spindle shape to a small round-to-polygonal cell, one of the characteristics of EFT. EWS/ETS also induced immunophenotypic changes in MPCs, including the disappearance of the mesenchyme-positive markers CD10 and CD13 and the up-regulation of the EFT-positive markers CD54, CD99, CD117, and CD271. Furthermore, a prominent shift from the gene expression profile of MPCs to that of EFT was observed in the presence of EWS/ETS. Together with the observation that EWS/ETS enhances the ability of cells to invade Matrigel, these results suggest that EWS/ETS proteins contribute to alterations of cellular features and confer an EFT-like phenotype to human MPCs.Ewing's family tumor (EFT) is a rare childhood cancer arising mainly in bone and soft tissue. Since EFT has a poor prognosis, it is important to elucidate the underlying pathogenic mechanisms for establishing a more effective therapeutic strategy. EFT is characterized by the presence of chimeric genes composed of EWS and ets transcription factor genes (ETS) formed by specific chromosomal translocations, i.e., EWS/FLI1, t(11;22)(q24;q12); EWS/ERG, t(21;22)(q12;q12); EWS/ETV1, t(7;22)(p22;q12); EWS/E1AF, t(17;22)(q12;q12); and EWS/FEV, t(2;22)(q33;q12) (26). The products of these chimeric genes behave as aberrant transcriptional regulators and are believed to play a crucial role in the onset and progression of EFT (3,36). Indeed, recent studies have revealed that the induction of EWS/FLI1 proteins can trigger transformation in certain cell types, including NIH 3T3 cells (36), C2C12 myoblasts (12), and murine primary bone marrow-derived mesenchymal progenitor cells (MPCs) (6, 45, 52). However, studies have also indicated that overexpression of EWS/ FLI1 provokes apoptosis and growth arrest in mouse normal embryonic fibroblasts and primary human fibroblasts (10, 31), hence hampering understanding of the precise role of EWS/ ETS proteins in the development of EFT. The function of EWS/ETS proteins would be greatly influenced by cell type, and thus the cells that can originate EFTs might be more susceptible to the tumorigenic effects of EWS/ETS.Although the cell origin of EFT is still unknown, the expression of neuronal markers in spite of the occurrence in bone and soft tissues has kept open the debate as to a potential mesenchymal or neuroectodermal origin. As described ...

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Section: Methodsmentioning

confidence: 99%

Inducible Expression of Chimeric EWS/ETS Proteins Confers Ewing's Family Tumor-Like Phenotypes to Human Mesenchymal Progenitor Cells

Miyagawa

Okita

Nakaijima

et al. 2008

Molecular and Cellular Biology

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“…Suppression of the expression of NPM inhibited proliferation of Ewing's sarcoma cells. The Ewing sarcoma cell lines, NCR‐EW2 , RD‐ES, and W‐ES , were transfected with NPM shRNAs or a negative control. (A) RT‐PCR analysis shows the decrease of NPM1 mRNA expression by two different shRNAs.…”

Section: Resultsmentioning

confidence: 99%

Interactomic approach for evaluating nucleophosmin‐binding proteins as biomarkers for Ewing's sarcoma

Haga

Ogawara²,

Kubota³

et al. 2013

Electrophoresis

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Nucleophosmin (NPM) is a novel prognostic biomarker for Ewing's sarcoma. To evaluate the prognostic utility of NPM, we conducted an interactomic approach to characterize the NPM protein complex in Ewing's sarcoma cells. A gene suppression assay revealed that NPM promoted cell proliferation and the invasive properties of Ewing's sarcoma cells. FLAG-tag-based affinity purification coupled with liquid chromatography-tandem mass spectrometry identified 106 proteins in the NPM protein complex. The functional classification suggested that the NPM complex participates in critical biological events, including ribosome biogenesis, regulation of transcription and translation, and protein folding, that are mediated by these proteins. In addition to JAK1, a candidate prognostic biomarker for Ewing's sarcoma, the NPM complex, includes 11 proteins known as prognostic biomarkers for other malignancies. Meta-analysis of gene expression profiles of 32 patients with Ewing's sarcoma revealed that 6 of 106 were significantly and independently associated with survival period. These observations suggest a functional role as well as prognostic value of these NPM complex proteins in Ewing's sarcoma. Further, our study suggests the potential applications of interactomics in conjunction with meta-analysis for biomarker discovery.

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“…3) Therefore, in order to diagnose these two tumors, several approaches are employed; macroscopic and microscopic analysis, ultrastructural analysis, and immunohistochemical analysis with specific antibodies to vimentin, desmin, neurofilaments, S-100, neuron-specific enolase, MIC2 4) and 5C11. 5) However, even such extensive analysis has failed to identify any specific morphological character of Ewing's sarcoma and PNET.…”

mentioning

confidence: 99%

“…5) However, even such extensive analysis has failed to identify any specific morphological character of Ewing's sarcoma and PNET.…”

mentioning

confidence: 99%

Molecular Analysis of Ewing's Sarcoma: Another Fusion Gene, EWS‐E1AF, Available for Diagnosis

Urano¹,

Umezawa²,

Yabe³

et al. 1998

Japanese Journal of Cancer Research

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Ewing's sarcoma, one of the most malignant tumors of children and young adults, expresses specific chimeric genes, e.g. EWS-FLI-1, EWS-ERG, EWS-ETV1 and EWS-FEV. In this paper, we extensively characterized a new fusion gene, EWS-E1AF by means of whole cDNA sequencing, RNA blot analysis, DNA blot analysis and chromosomal analysis, and showed it to be available for the diagnosis of Ewing's sarcoma and to participate in the oncogenesis of Ewing's sarcoma. Furthermore, we conducted a genetic analysis of Ewing family tumors in conjuction with immunohistochemical analysis and ultrastructural analysis. Our results demonstrate some limitations of both genetic analysis and histopathological analysis, and establish the relationship between neurogenic phenotypes and chimera genes.Key words: Ewing's sarcoma -Molecular diagnosis -E1AF -Ets -RNA binding protein Ewing's sarcoma, which occurs in the bone and soft tissue, is one of the most malignant tumors in young people.1) This tumor does not have specific morphological phenotypes. By light microscopic analysis, it is sometimes difficult to differentiate Ewing's sarcoma from neuroblastoma, malignant lymphoma, some types of rhabdomyosarcoma and osteosarcoma.2) Similarly, peripheral primitive neuroectodermal tumor (PNET), which has similar morphology but has neuronal character, is also hard to diagnose.3) Therefore, in order to diagnose these two tumors, several approaches are employed; macroscopic and microscopic analysis, ultrastructural analysis, and immunohistochemical analysis with specific antibodies to vimentin, desmin, neurofilaments, S-100, neuron-specific enolase, MIC2 4) and 5C11. 5) However, even such extensive analysis has failed to identify any specific morphological character of Ewing's sarcoma and PNET.Many human tumors contain characteristic chromosomal translocations. Through the chromosomal translocations, specific fusion genes are generated.6, 7) These fusion genes are considered to play an important role in oncogenesis. Ewing' sarcoma and PNET have the same specific chromosomal translocation, t(11;22) 8,9) and through this translocation, the EWS-FLI-1 fusion gene is generated.10) FLI-1, a partner of the fusion gene, is a member of the Ets family of transcription factors. Other Ets family genes, ERG and ETV1, are also fused to EWS through t(21;22) and t(7;22), respectively. 11-13) Recently, we identified a new fusion gene, EWS-E1AF in a case of Ewing's sarcoma.14) Since analysis of the fusion genes revealed that Ewing's sarcoma and PNET have the same fusion transcripts, Ewing's sarcoma and PNET are considered to belong to a common entity. 15) In this study, we extensively characterized the EWS-E1AF fusion gene in the established cell line NCR-EW3, which has t(17;22) (q21;q21). Furthermore, we conducted a genetic analysis of Ewing's sarcoma and PNET in comparison with the accumulated data on clinical history and the results of pathological examination. MATERIALS AND METHODSTumors and cell lines Thirty surgical samples were obtained from Japanese patients. On ...

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A monoclonal antibody specifically reactive with Ewing's sarcoma

Cited by 26 publications

References 7 publications

Inducible Expression of Chimeric EWS/ETS Proteins Confers Ewing's Family Tumor-Like Phenotypes to Human Mesenchymal Progenitor Cells

Inducible Expression of Chimeric EWS/ETS Proteins Confers Ewing's Family Tumor-Like Phenotypes to Human Mesenchymal Progenitor Cells

Interactomic approach for evaluating nucleophosmin‐binding proteins as biomarkers for Ewing's sarcoma

Molecular Analysis of Ewing's Sarcoma: Another Fusion Gene, EWS‐E1AF, Available for Diagnosis

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