scholarly journals γ-Glutamyl transpeptidase expression in Ewing's sarcoma cells:up-regulation by interferons

2002 ◽  
Vol 364 (3) ◽  
pp. 719-724 ◽  
Author(s):  
Lena BOUMAN ◽  
Josiane SANCÉAU ◽  
Dany ROUILLARD ◽  
Brigitte BAUVOIS
Keyword(s):  
Cell Lines ◽  
Ewing’S Sarcoma ◽  
Ewing's Sarcoma ◽  
Fusion Gene ◽  
Chromosomal Translocation ◽  
Cellular Transformation ◽  
Enzymic Activity ◽  
Type I ◽  
Chromosomal Breaks ◽  
Glutamyl Transpeptidase

The genetic hallmark of Ewing's sarcoma family of tumours (ET) is the presence of the translocation t(11;22)(q24;q12), which creates the ET fusion gene, leading to cellular transformation. Five human γ-glutamyl transpeptidase (γ-GT) genes are located near the chromosomal translocation in ET. γ-GT is a major enzyme involved in glutathione homoeostasis. Five human cell lines representative of primary or metastatic tumours were investigated to study whether γ-GT alterations could occur at the chromosomal breaks and rearrangements in ET. As shown by enzymic assays and FACS analyses, all ET cell lines consistently expressed a functional γ-GT which however did not discriminate steps of ET progression. As shown previously [Sancéau, Hiscott, Delattre and Wietzerbin (2000) Oncogene 19, 3372–3383], ET cells respond to the antiproliferative effects of interferons (IFNs) type I (α and β) and to a much less degree to IFN type II (γ). IFN-α and -β arrested cells in the S-phase of the cell cycle. We found an enhancement of γ-GT mRNA species with IFN-α and -β by reverse transcriptase—PCR analyses. This is reflected by up-regulation of γ-GT protein, which coincides with the increase in γ-GT-specific enzymic activity. Similarly, IFNs up-regulate the levels of γ-GT in another IFN-responsive B cell line. Whether this up-regulation of γ-GT by IFNs is of physiological relevance to cell behaviour remains to be studied.

scholarly journals Identification of target genes for the Ewing's sarcoma EWS/FLI fusion protein by representational difference analysis.

1995 ◽  
Vol 15 (8) ◽  
pp. 4623-4630 ◽  
Author(s):  
B S Braun ◽  
R Frieden ◽  
S L Lessnick ◽  
W A May ◽  
C T Denny
Keyword(s):  
Target Genes ◽  
Ewing’S Sarcoma ◽  
Ewing's Sarcoma ◽  
Germ Line ◽  
Fusion Gene ◽  
Chromosomal Translocation ◽  
Representational Difference Analysis ◽  
Difference Analysis ◽  
Broad Array ◽  
Direct Target Gene

The EWS/FLI-1 fusion gene results from the 11;22 chromosomal translocation in Ewing's sarcoma. The product of the gene is one of a growing number of structurally altered transcription factors implicated in oncogenesis. We have employed a subtractive cloning strategy of representational difference analysis in conjunction with a model transformation system to identify genes transcribed in response to EWS/FLI. We have characterized eight transcripts that are dependent on EWS/FLI for expression and two transcripts that are repressed in response to EWS/FLI. Three of the former were identified by sequence analysis as stromelysin 1, a murine homolog of cytochrome P-450 F1 and cytokeratin 15. Stromelysin 1 is induced rapidly after expression of EWS/FLI, suggesting that the stromelysin 1 gene may be a direct target gene of EWS/FLI. These results demonstrate that expression of EWS/FLI leads to significant changes in the transcription of specific genes and that these effects are at least partially distinct from those caused by expression of germ line FLI-1. The representational difference analysis technique can potentially be applied to investigate transformation pathways activated by a broad array of genes in different tumor systems.

scholarly journals Characterization and Drug Resistance Patterns of Ewing's Sarcoma Family Tumor Cell Lines

PLoS ONE ◽  
2013 ◽  
Vol 8 (12) ◽  
pp. e80060 ◽  
Author(s):  
William A. May ◽  
Rita S. Grigoryan ◽  
Nino Keshelava ◽  
Daniel J. Cabral ◽  
Laura L. Christensen ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Tumor Cell ◽  
Cell Lines ◽  
Ewing’S Sarcoma ◽  
Ewing's Sarcoma ◽  
Tumor Cell Lines ◽  
Resistance Patterns

scholarly journals 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.

1993 ◽  
Vol 13 (12) ◽  
pp. 7393-7398 ◽  
Author(s):  
W A May ◽  
S L Lessnick ◽  
B S Braun ◽  
M Klemsz ◽  
B C Lewis ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Primitive Neuroectodermal Tumor ◽  
Ewing’S Sarcoma ◽  
Ewing's Sarcoma ◽  
Fusion Gene ◽  
Consensus Sequence ◽  
Transcriptional Activator ◽  
Neuroectodermal Tumor ◽  
Amino Terminal ◽  
Transforming Gene

EWS/FLI-1 is a chimeric protein formed by a tumor-specific 11;22 translocation found in both Ewing's sarcoma and primitive neuroectodermal tumor of childhood. EWS/FLI-1 has been shown to be a potent transforming gene, suggesting that it plays an important role in the genesis of these human tumors. We now demonstrate that EWS/FLI-1 has the characteristics of an aberrant transcription factor. Subcellular fractionation experiments localized the EWS/FLI-1 protein to the nucleus of primitive neuroectodermal tumor cells. EWS/FLI-1 specifically bound in vitro an ets-2 consensus sequence similarly to normal FLI-1. When coupled to a GAL4 DNA-binding domain, the amino-terminal EWS/FLI-1 region was a much more potent transcriptional activator than the corresponding amino-terminal domain of FLI-1. Finally, EWS/FLI-1 efficiently transformed NIH 3T3 cells, but FLI-1 did not. These data suggest that EWS/FLI-1, functioning as a transcription factor, leads to a phenotype dramatically different from that of cells expressing FLI-1. EWS/FLI-1 could disrupt normal growth and differentiation either by more efficiently activating FLI-1 target genes or by inappropriately modulating genes normally not responsive to FLI-1.

scholarly journals An Innovative Therapeutic Option for the Treatment of Skeletal Sarcomas: Elimination of Osteo- and Ewing’s Sarcoma Cells Using Physical Gas Plasma

2020 ◽  
Vol 21 (12) ◽  
pp. 4460 ◽  
Author(s):  
Josephine M. Jacoby ◽  
Silas Strakeljahn ◽  
Andreas Nitsch ◽  
Sander Bekeschus ◽  
Peter Hinz ◽  
...  
Keyword(s):  
Cell Membrane ◽  
Cell Lines ◽  
Ewing’S Sarcoma ◽  
Ewing's Sarcoma ◽  
Therapeutic Option ◽  
Bone Sarcoma ◽  
Atmospheric Plasma ◽  
Local Surgery ◽  
Release Assay ◽  
Sarcoma Cells

Osteosarcoma and Ewing’s sarcoma are the most common malignant bone tumors. Conventional therapies such as polychemotherapy, local surgery, and radiotherapy improve the clinical outcome for patients. However, they are accompanied by acute and chronic side effects that affect the quality of life of patients, motivating novel research lines on therapeutic options for the treatment of sarcomas. Previous experimental work with physical plasma operated at body temperature (cold atmospheric plasma, CAP) demonstrated anti-oncogenic effects on different cancer cell types. This study investigated the anti-cancer effect of CAP on two bone sarcoma entities, osteosarcoma and Ewing’s sarcoma, which were represented by four cell lines (U2-OS, MNNG/HOS, A673, and RD-ES). A time-dependent anti-proliferative effect of CAP on all cell lines was observed. CAP-induced alterations in cell membrane functionality were detected by performing a fluorescein diacetate (FDA) release assay and an ATP release assay. Additionally, modifications of the cell membrane and modifications in the actin cytoskeleton composition were examined using fluorescence microscopy monitoring dextran-uptake assay and G-/F-actin distribution. Furthermore, the CAP-induced induction of apoptosis was determined by TUNEL and active caspases assays. The observations suggest that a single CAP treatment of bone sarcoma cells may have significant anti-oncogenic effects and thus may be a promising extension to existing applications.

scholarly journals Inhibition of heat shock proteins (HSP) expression by quercetin and differential doxorubicin sensitization in neuroblastoma and Ewing’s sarcoma cell lines

2007 ◽  
Vol 103 (4) ◽  
pp. 1344-1354 ◽  
Author(s):  
Cristina Zanini ◽  
Giuliana Giribaldi ◽  
Giorgia Mandili ◽  
Franco Carta ◽  
Nicoletta Crescenzio ◽  
...  
Keyword(s):  
Heat Shock ◽  
Heat Shock Proteins ◽  
Cell Lines ◽  
Ewing’S Sarcoma ◽  
Ewing's Sarcoma ◽  
Sarcoma Cell ◽  
Shock Proteins

scholarly journals Discovery and Validation of a Compound to Target Ewing’s Sarcoma

Pharmaceutics ◽  
2021 ◽  
Vol 13 (10) ◽  
pp. 1553
Author(s):  
Ellie Esfandiari Nazzaro ◽  
Fahad Y. Sabei ◽  
Walter K. Vogel ◽  
Mohamad Nazari ◽  
Katelyn S. Nicholson ◽  
...  
Keyword(s):  
Cell Lines ◽  
Ewing’S Sarcoma ◽  
Ewing's Sarcoma ◽  
Apoptotic Cell ◽  
Intensive Therapy ◽  
Xenograft Model ◽  
Chemotherapeutic Agents ◽  
Cytotoxic Agents ◽  
Sarcoma Cell

Ewing’s sarcoma, characterized by pathognomonic t (11; 22) (q24; q12) and related chromosomal ETS family translocations, is a rare aggressive cancer of bone and soft tissue. Current protocols that include cytotoxic chemotherapeutic agents effectively treat localized disease; however, these aggressive therapies may result in treatment-related morbidities including second-site cancers in survivors. Moreover, the five-year survival rate in patients with relapsed, recurrent, or metastatic disease is less than 30%, despite intensive therapy with these cytotoxic agents. By using high-throughput phenotypic screening of small molecule libraries, we identified a previously uncharacterized compound (ML111) that inhibited in vitro proliferation of six established Ewing’s sarcoma cell lines with nanomolar potency. Proteomic studies show that ML111 treatment induced prometaphase arrest followed by rapid caspase-dependent apoptotic cell death in Ewing’s sarcoma cell lines. ML111, delivered via methoxypoly(ethylene glycol)-polycaprolactone copolymer nanoparticles, induced dose-dependent inhibition of Ewing’s sarcoma tumor growth in a murine xenograft model and invoked prometaphase arrest in vivo, consistent with in vitro data. These results suggest that ML111 represents a promising new drug lead for further preclinical studies and is a potential clinical development for the treatment of Ewing’s sarcoma.

scholarly journals PDX-Derived Ewing’s Sarcoma Cells Retain High Viability and Disease Phenotype in Alginate Encapsulated Spheroid Cultures

Cancers ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 879
Author(s):  
Giacomo Domenici ◽  
Rodrigo Eduardo ◽  
Helena Castillo-Ecija ◽  
Gorka Orive ◽  
Ángel Montero Carcaboso ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Lines ◽  
Drug Sensitivity ◽  
Ewing’S Sarcoma ◽  
Ewing's Sarcoma ◽  
3D Cell Culture ◽  
Suitable Model ◽  
Es Cell ◽  
3D Cultures ◽  
Novel Drugs

Ewing’s Sarcoma (ES) is the second most frequent malignant bone tumour in children and young adults and currently only untargeted chemotherapeutic approaches and surgery are available as treatment, although clinical trials are on-going for recently developed ES-targeted therapies. To study ES pathobiology and develop novel drugs, established cell lines and patient-derived xenografts (PDX) are the most employed experimental models. Nevertheless, the establishment of ES cell lines is difficult and the extensive use of PDX raises economic/ethical concerns. There is a growing consensus regarding the use of 3D cell culture to recapitulate physiological and pathophysiological features of human tissues, including drug sensitivity. Herein, we implemented a 3D cell culture methodology based on encapsulation of PDX-derived ES cell spheroids in alginate and maintenance in agitation-based culture systems. Under these conditions, ES cells displayed high proliferative and metabolic activity, while retaining the typical EWSR1-FLI1 chromosomal translocation. Importantly, 3D cultures presented reduced mouse PDX cell contamination compared to 2D cultures. Finally, we show that these 3D cultures can be employed in drug sensitivity assays, with results similar to those reported for the PDX of origin. In conclusion, this novel 3D cell culture method involving ES-PDX-derived cells is a suitable model to study ES pathobiology and can assist in the development of novel drugs against this disease, complementing PDX studies.

Sign in / Sign up

Export Citation Format

Share Document