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| Status |
Public on Oct 01, 2008 |
| Title |
EZH2 is a mediator of EWS-FLI1 driven tumor growth blocking endothelial and neuro-ectodermal differentiation |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
Ewing Tumors (ET) are highly malignant tumors, localized in bone or soft tissue and are molecularly defined by ews/ets translocations. We identified histone methyl-transferase Enhancer of Zeste, Drosophila, Homolog 2 (EZH2) to be increased in ET. EZH2’s suppressive activity maintains stemness in normal and malignant cells. Here we found EZH2 to be upregulated by the pathognomonic fusion oncogene EWS-FLI1 in ET and mesenchymal stem cells. Downregulation of EZH2 by RNA interference in ET suppressed oncogenic transformation by inhibiting clonogenicity in vitro. Similarly, tumor development and metastasis in immunodeficient Rag2-/-γC-/- mice was suppressed. EZH2-mediated gene silencing was shown to be dependent on histone deacetylase (HDAC) activity. Subsequent microarray analysis of EZH2 knock down, HDAC-inhibitor treatment and confirmation in independent assays revealed an undifferentiated phenotype maintained by EZH2 in ET. Downregulation of EZH2 decreased histone H3 lysine 27 trimethylation (H3K27me3) at target loci. EZH2 regulated stemness genes such as nerve growth factor receptor (NGFR) as well as genes involved in neuroectodermal differentiation (EMP1, EPHB2, GFAP, GAP43). These data suggest that EZH2 might play a central role in Ewing Tumor pathology shaping the oncogenicity and stem cell phenotype of this tumor presumably by epigenetic regulation.
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| Overall design |
A673 cells were treated for 24 hours either with 100 nM Trichostatin A (TSA) or 0.01% DMSO. In siRNA experiments with ezh2 specific siRNA A673 cells were resuspended in medium containing 5 nM siRNA and transfection reagent and incubated for 48 hours. RNA from cells under such treatment was isolated with Trizol and subjected to microarray analysis onto human U133A microarray following the Affymetrix protocol.
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| Contributor(s) |
Richter GH, Roessler S, Staege MS, Burdach S |
| Citation(s) |
19289832 |
| |
| Submission date |
Sep 07, 2008 |
| Last update date |
Aug 10, 2018 |
| Contact name |
Guenther HS Richter |
| E-mail(s) |
guenther.richter@charite.de
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| Organization name |
Charité - Universitätsmedizin Berlin
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| Department |
Department of Pediatrics, Division of Oncology and Hematology
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| Street address |
Augustenburger Platz 1
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| City |
Berlin |
| ZIP/Postal code |
D-13353 |
| Country |
Germany |
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| Platforms (1) |
| GPL96 |
[HG-U133A] Affymetrix Human Genome U133A Array |
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| Samples (9)
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| GSM318628 |
A673 cells treated with control siRNA, biological rep2 |
| GSM318629 |
A673 cells treated with DMSO, biological rep2 |
| GSM318630 |
A673 cells treated with TSA, biological rep2 |
| GSM318631 |
A673 cells treated with ezh2 siRNA no.2, biological rep2 |
| GSM318632 |
A673 cells treated with control siRNA, biological rep1 |
| GSM318633 |
A673 cells treated with DMSO, biological rep1 |
| GSM318634 |
A673 cells treated with TSA, biological rep1 |
| GSM318635 |
A673 cells treated with ezh2 siRNA no.2, biological rep1 |
| GSM318636 |
A673 cells treated with ezh2 siRNA-validated |
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| Relations |
| BioProject |
PRJNA112701 |
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