Effect of radiation on Ewing tumour subpopulations characterized on a single-cell level: intracellular cytokine, immunophenotypic, DNA and apoptotic profile

Könemann, Stefan; Bölling, Tobias; Schuck, Andreas; Malath, J.; Kolkmeyer, Astrid; Horn, Kirsten; Riesenbeck, D.; Hesselmann, Stefan; Diallo, Raihanatou; Vormoor, Josef; Willich, Normann

doi:10.1080/0955300031000073400

Cited by 4 publications

(7 citation statements)

References 48 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Notably, the expression of CD54 (intercellular cell adhesion molecule-1 [ICAM1]), CD117 (c-kit), and CD271 (low-affinity nerve growth factor receptor [LNGFR]) increased in EWS/ETS-expressing UET-13 cells. These markers are positive in EFT cell lines (17,28,33), and in addition, CD117 is detected in about 40% of patient samples (17) and is negative in human primary MPCs (4,43). Thus, it is reasonable to consider that a phenotypic marker of EFT was induced in UET-13 cells by EWS/ETS expression.…”

Section: Discussionmentioning

confidence: 99%

“…Human MPCs reveal a characteristic expression of several surface antigens and can be identified on the basis of the reactivity with a set of monoclonal antibodies against CD antigens (25,42). On the other hand, some CD antigens are characteristically expressed on EFT cells (17,28,33). Using the combinations of these antibodies listed in Table 2, which are useful for the immunodetection of either MPCs or EFT cells, we further examined whether EWS/ETS expression affects the immunophenotype of UET-13 cells and compared its effect with that on the immunophenotype of EFT cell lines (Table 2 and Fig.…”

Section: Vol 28 2008 Effects Of Ews/ets Expression In Human Mpcs 2129mentioning

confidence: 99%

See 1 more Smart Citation

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

View full text Add to dashboard Cite

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 ...

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Vol 28 2008 Effects Of Ews/ets Expression In Human Mpcs 2129mentioning

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

View full text Add to dashboard Cite

show abstract

“…14 We now focused on the markers HLA-ABC, CD11a and CD62L because our results indicated that these markers might be informative to detect heterogeneity in view of radiation induced apoptosis. These markers are described to be involved in apoptosis related pathways and in the clearance process of apoptotic cells.…”

Section: Introductionmentioning

confidence: 95%

“…A simultaneous characterization of light scatter properties, surface molecules, DNA content, and apoptosis-associated markers can be performed at a single cell level. [11][12][13][14] As an early marker of apoptosis the binding of Annexin V to phosphatidylserine on the cell membrane can be used. 15 In addition, the staining of the DNA with dyes like 7-AAD without any permeabilisation of the cell membrane allows the characterization of the integrity of the cell membrane and is therefore a useful marker for characterization of apoptosis.…”

Section: Introductionmentioning

confidence: 99%

Heterogeneity of radiation induced apoptosis in Ewing Tumor cell lines characterized on a single cell level

et al. 2005

Self Cite

View full text Add to dashboard Cite

The objective of this study was to investigate heterogeneity of radiation induced apoptosis on a single cell level. Two Ewing tumor cell lines were characterized in vitro before and 24 and 72 h after radiation with 5 Gy by multiparametric flow cytometry. Annexin V, 7-AAD and fluorescence conjugated antibodies that were directed against HLA-ABC, CD11a and CD62L were used. Based on these markers radiation induced apoptosis was quantified, multiple apoptotic subpopulations were identified and a characteristic individual apoptotic profile was characterized. The characterization of HLA-ABC, CD11a and CD62L was informative to detect subpopulations of apoptotic cells. The observed heterogeneity and the identification of multiple apoptotic subpopulations reflect the complexity and diversity of biology of radiation induced cell death. This might be an indication for co-existing apoptotic pathways or it might represent sequential steps of the apoptotic cascade.

show abstract

“…Multiparametric flow cytometry is an effective, reliable and fast technique to characterize tumour cells and identify interindividual heterogeneity and subpopulations within one individual tumour. Using this method, the simultaneous characterization of cell surface antigens, intracellular cytokines, DNA content and apoptosis-associated markers can be used at a single cell level (Hollander and Loken 1988, Brons et al 1990, Stokke et al 1991, Orfao et al 1995, Prussin and Metcalfe 1995, Corver et al 1996, Jason and Larned 1997, Könemann et al 2000, 2003, Bacso and Eliason 2001, Rasola and Geuna 2001.…”

Section: Introductionmentioning

confidence: 99%

Time‐ and dose‐dependent changes of intracellular cytokine and cytokine receptor profile of Ewing tumour subpopulations under the influence of ionizing radiation

Könemann

Bölling

Malath

et al. 2003

International Journal of Radiation Biology

Self Cite

View full text Add to dashboard Cite

The observed radiation-induced changes of cytokine and receptor profile are an indication for complex intercellular interactions in view of radioresistance-associated mechanisms between cell populations within one individual tumour. The observed heterogeneous response on radiation might have therapeutic implications for an individualized therapy based on combined radiation and cytokine modulation, defined by flow cytometric characterization of markers potentially informative for radioresistance.

show abstract

Effect of radiation on Ewing tumour subpopulations characterized on a single-cell level: intracellular cytokine, immunophenotypic, DNA and apoptotic profile

Cited by 4 publications

References 48 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

Heterogeneity of radiation induced apoptosis in Ewing Tumor cell lines characterized on a single cell level

Time‐ and dose‐dependent changes of intracellular cytokine and cytokine receptor profile of Ewing tumour subpopulations under the influence of ionizing radiation

Contact Info

Product

Resources

About