Reverse Transcriptase PCR Amplification of EWS/FLI-1 Fusion Transcripts as a Diagnostic Test for Peripheral Primitive Neuroectodermal Tumors of Childhood

Sorensen, Poul H.; Liu, Xian Fang; Delattre, Olivier; Rowland, Jon M.; Biggs, Christopher; Thomas, Gilles; Triche, Timothy J.

doi:10.1097/00019606-199309000-00002

Cited by 135 publications

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“…We postulated that this effect might be mediated through either the Ras-ERK1/2 or PI3K-Akt cascade because both become activated in ET spheroids (25). To test this, we used two previously characterized ET cell lines, TC32 and TC71, each of which expresses the ET-specific EWS-FLI1 gene fusion (25,27). TC32 and TC71 monolayer and spheroid cultures were assessed for caspase-3 activation (to measure apoptosis) after treatment with the PI3K inhibitor LY294002 or the MEK inhibitor U0126.…”

Section: Resultsmentioning

confidence: 99%

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E-Cadherin Cell-Cell Adhesion in Ewing Tumor Cells Mediates Suppression of Anoikis through Activation of the ErbB4 Tyrosine Kinase

et al. 2007

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Ability to grow under anchorage-independent conditions is one of the major hallmarks of transformed cells. Key to this is the capacity of cells to suppress anoikis, or programmed cell death induced by detachment from the extracellular matrix. To model this phenomenon in vitro, we plated Ewing tumor cells under anchorage-independent conditions by transferring them to dishes coated with agar to prevent attachment to underlying plastic. This resulted in marked up-regulation of E-cadherin and rapid formation of multicellular spheroids in suspension. Addition of calcium chelators, antibodies to E-cadherin (but not to other cadherins or B 1 -integrin), or expression of dominant negative E-cadherin led to massive apoptosis of spheroid cultures whereas adherent cultures were unaffected. This correlated with reduced activation of the phosphatidylinositol 3-kinase-Akt pathway but not the Rasextracellular signal-regulated kinase 1/2 cascade. Furthermore, spheroid cultures showed profound chemoresistance to multiple cytotoxic agents compared with adherent cultures, which could be reversed by A-E-cadherin antibodies or dominant negative E-cadherin. In a screen for potential downstream effectors of spheroid cell survival, we detected E-cadherin-dependent activation of the ErbB4 receptor tyrosine kinase but not of other ErbB family members. Reduction of ErbB4 levels by RNA interference blocked Akt activation and spheroid cell survival and restored chemosensitivity to Ewing sarcoma spheroids. Our results indicate that anchorage-independent Ewing sarcoma cells suppress anoikis through a pathway involving E-cadherin cell-cell adhesion, which leads to ErbB4 activation of the phosphatidylinositol 3-kinase-Akt pathway, and that this is associated with increased resistance of cells to cytotoxic agents. [Cancer Res 2007;67(7):3094-105]

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Section: Resultsmentioning

confidence: 99%

“…TC32 and TC71 ET cell lines and their growth requirements have previously been described (27). For anchorageindependent (spheroid) cultures, monolayer cells were trypsinized, resuspended as single cells, and replated at a concentration of 3.0 Â 10 5 /mL on standard dishes coated with 1.4% agar as described (25,28).…”

Section: Methodsmentioning

confidence: 99%

E-Cadherin Cell-Cell Adhesion in Ewing Tumor Cells Mediates Suppression of Anoikis through Activation of the ErbB4 Tyrosine Kinase

et al. 2007

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“…Other rarer translocations include t(7;22)(p22; q12) resulting in EWS/ETV-1; t(17;22) (q12; q12) resulting in EWS/E1AF and t(2;22)(q33; q12) resulting in EWS/FEV (40). EWS/FL-1 fusion transcript caused by t(11;22)(q24; q12) is specific for ES/PNET and is not found in other small round cell tumors of infancy (41,42). Moreover, this translocation can be demonstrated in up to 80% of ES/PNET by reverse transcriptase-PCR from mRNA isolated from fresh or formalin-fixed, paraffin-embedded tissue (41)(42)(43)(44).…”

Section: Discussionmentioning

confidence: 99%

“…EWS/FL-1 fusion transcript caused by t(11;22)(q24; q12) is specific for ES/PNET and is not found in other small round cell tumors of infancy (41,42). Moreover, this translocation can be demonstrated in up to 80% of ES/PNET by reverse transcriptase-PCR from mRNA isolated from fresh or formalin-fixed, paraffin-embedded tissue (41)(42)(43)(44). Interestingly, one reported case of acute myeloid leukemia had t(11;22)(q24; q12) translocation by cytogenetic analysis (45), but it is not known whether that case contained EWS/FL-1 fusion transcript, because the case was not studied at the molecular level.…”

Section: Discussionmentioning

confidence: 99%

Differentiating Lymphoblastic Lymphoma and Ewing's Sarcoma: Lymphocyte Markers and Gene Rearrangement

et al. 2001

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We encountered a child with an intraosseous small round cell tumor that was negative for LCA, CD20 (L26), and CD3 and positive for vimentin, CD99 (MIC-2), and periodic acid-Schiff. The tumor exhibited rosette-like formations. This case was initially interpreted as Ewing's sarcoma (ES); however, additional studies revealed positivity for CD79a, CD43, and TdT expression, and an immunoglobulin heavy chain gene rearrangement (IgH-R) by polymerase chain reaction (PCR) established this to be a precursor B-lymphoblastic lymphoma. Because the differential diagnosis of ES and lymphoblastic lymphoma can be difficult and the differential diagnostic value of leukocyte antigens and immunoglobulin heavy chain gene rearrangement studies have not been fully evaluated, we conducted a more extensive investigation on 33 (21 soft tissue and 12 intraosseous) ES cases. Cases were retrieved from the files of the Department of Pathology at Georgetown University and from the Soft Tissue Registry of the Armed Forces Institute of Pathology. The cases were studied by light microscopy, immunohistochemistry, and PCR for IgH-R and T cell receptor gamma chain gene rearrangement (T␥-R). There were 17 females and 16 males; the mean age was 29.3 years. Locations included the extremities (n ‫؍‬ 17) and trunk (n ‫؍‬ 16). All cases fit the ES spectrum by light microscopy and immunohistochemistry, as previously determined, and were negative for lymphoid markers (LCA, CD3, CD20, CD43, CD79a, and TdT), CD10 and CD34. CD99 was positive in 31/33 and bcl-2 was weakly positive in 13/33 cases. All 21 cases studied for gene rearrangements by PCR were negative for IgH-R and T␥-R. Distinction of intraosseous lymphoblastic lymphoma from ES may be difficult because lymphomas may occasionally exhibit unexpected morphologic and immunophenotypic properties including LCA, CD3 and CD20 negativity and cytokeratin positivity. Additional analysis using CD79a, CD43, TdT, and PCR should be performed to avoid misdiagnosis. True ES is negative for lymphoid markers including CD79a, CD43, and TdT, as well as for IgH-R and T␥-R.

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“…Primitive neural ectodermal tumors (PNET) are similar to Ewing sarcoma and share the frequent t(11;22). Gorman et al 23 and Sorenson et al 24 have both reported PNET cell lines with normal cytogenetics.…”

Section: Discussionmentioning

confidence: 99%

Characterization of 11 human sarcoma cell strains

Nicolson

Trent

et al. 2002

Cancer

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BACKGROUNDHuman sarcomas have a propensity for aggressive local invasion and early pulmonary metastasis. Frequently, deaths are due to uncontrolled pulmonary metastases. The purpose of the current study was to evaluate cytogenetics, tumorigenicity, metastatic potential, and production of angiogenic factors in human sarcoma cell strains. A secondary purpose was to establish low passage cell strains for studying new therapeutic approaches.METHODSThe authors established 11 cell strains from human sarcoma surgical specimens and characterized their in vitro tumor properties, including growth in soft agar, expression of angiogenic growth factors (vascular endothelial growth factor [VEGF] and basic‐fibroblast growth factor [bFGF]), and cytogenetics.RESULTSAll of the cell strains remained diploid. All exhibited the ability to grow in soft agar and expressed both VEGF as well as bFGF. In addition, 6 of the 11 established sarcoma cell strains were tumorigenic, 5 of which spontaneously metastasized to the lungs in nude mice. Four of the five cell strains that yielded lung metastases were derived from lung metastases in patients.CONCLUSIONSThe 11 cell strains, which were derived from diverse sarcoma histologies, will provide a model for studying not only metastatic progression but also the in vitro and in vivo efficacy of new therapeutic modalities for human sarcomas. Cancer 2002;95:1569–76. © 2002 American Cancer Society.DOI 10.1002/cncr.10879

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Reverse Transcriptase PCR Amplification of EWS/FLI-1 Fusion Transcripts as a Diagnostic Test for Peripheral Primitive Neuroectodermal Tumors of Childhood

Cited by 135 publications

References 0 publications

E-Cadherin Cell-Cell Adhesion in Ewing Tumor Cells Mediates Suppression of Anoikis through Activation of the ErbB4 Tyrosine Kinase

E-Cadherin Cell-Cell Adhesion in Ewing Tumor Cells Mediates Suppression of Anoikis through Activation of the ErbB4 Tyrosine Kinase

Differentiating Lymphoblastic Lymphoma and Ewing's Sarcoma: Lymphocyte Markers and Gene Rearrangement

Characterization of 11 human sarcoma cell strains

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