Abstract
We investigated the capacity of eight well-characterized Ewing's sarcoma cell lines to differentiate towards a neural phenotype. Ewing's sarcoma cells expressed the neuroectoderm-associated antigens such as nerve growth factor (NGF) receptor, N-CAM (6H7 and Leu-19) and Leu-7. Ewing's sarcoma cells also exhibited the potential for neural differentiation at the mRNA level; neuron-specific medium- and low-sized filament (NF-M and NF-L) expression was induced by dibutyryladenosine cyclic monophosphate. The pattern of expression of NF-L obtained by using alternative polyadenylation sites in Ewing's sarcoma cells differed from that in peripheral primitive neuroectodermal tumour (PNET) cells, and was similar to that in undifferentiated neural tissues. Furthermore, the NGF receptors detected by immunohistochemistry were found to be non-functional as assayed byc-fos induction with NGF treatment. The results indicate that Ewing's sarcoma cells maintain a primitive phenotype and have the potential to differentiate into a neural phenotype, indicating that Ewing's sarcoma is distinct from PNET.
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Bosselut RA, Zucman J, Cormier F, Delattre O, Roussel M, Thomas G, Ghysdael J (1994) DNA-binding and transcriptional activation properties of the EWS-FLI-1 fusion protein resulting from the t(11;22) translocation in Ewing sarcoma. Mol Cell Biol 14:3230–3241
Baker DL, Reddy DR, Pleasure D, Thorpe CL, Evans AE, Cohen PS, Ross AH (1989) Analysis of nerve growth factor receptor expression in human neuroblastoma and neuroepithelioma cell lines. Cancer Res 49:4142–4146
Barbas JA, Chaix JC, Steinmetz M, Goridis C (1988) Differential splicing and alternative polyadenylation generates distinct NCAM transcripts and proteins in the mouse. EMBO J 7:625–632
Ben-David Y, Giddens EB, Letwin K, Bernstein A (1991) Erythroleukemia induction by Friend murine leukemia virus: insertional activation of a new member of the ets gene family, Fli-1, closely linked to c-ets-1. Genes Dev 5:908–918
Bennett J, Catovsky D (1982) Proposals for the classification of the myelodysplastic syndromes. Br J Haematol 51:189–199
Bloom E (1972) Further definition by cytotoxicity tests of cell surface antigens of human sarcomas in culture. Cancer Res 32:960–967
Carden MJ, Trojanowski JQ, Schlaepfer WW, Lee V-Y (1987) Two-stage expression of neurofilament polypeptides during rat neurogenesis with early establishment of adult phosphorylation patterns. J Neurosci 7:3489–3504
Cavazzana AO, Miser JC, Jefferson J, Triche TJ (1987) Experimental evidence for neural origin of Ewing's sarcoma of bone. Am J Pathol 127:507–518
Delmer LP (1993) Primitive neuroectodermal tumour and Swing's sarcoma. Am J Surg Pathol 17:1–13
Delattre O, Zucman J, Plougastel B, Desmeze C, Melot T, Peter M, Kovar H, Joubert I, de Jong P, Rouleau G, Aurias A, Thomas G (1992) Gene fusion with an ETS DNA-binding domain caused by chromosome translocation in human tumours. Nature 359:162–165
Delattre O, Zucmann J, Melot T, Garau XS, Zucker J, Lenoir GM, Ambros PF, Sheer D, Turc-Carel C, Triche TJ, Aurias A, Thomas G (1994) The Swing family of tumours — a subgroup of small-round-cell tumours defined by specific chimeric transcripts. New Engl J Med 331:294–299
Dockhorn-Dworniczak B, Schäfer K-L, Dantcheva R, Blasius S, Winkelmann W, Strehl S, Burdach S, Valen F, Jürgens H, Böcker W (1994) Diagnostic value of the molecular genetic detection of the t(11;22) translocation in Swing's tumours. Virchows Arch 425:107–112
Enzinger FM, Weiss SW (1988) Soft tissue tumour. C.V. Mosby, St. Louis, pp 1–18, 806–815, 951–958
Swing J (1921) Diffuse endothelioma of bone. Proc NY Pathol Soc 21:17–24
Fujii Y, Hongo T, Nakagawa Y, Nasuda K, Mizuno Y, Igarashi Y, Naito Y, Maeda M (1989) Cell culture of small round cell tumour originating in the thoracopulmonary resion. Evidence for derivation from a primitive pluripotent cell. Cancer 64:43–51
Gardinier MV, Macklin WB, Diniak AJ, Deininger PL (1986) Characterization of myelin proteolipid mRNAs in normal and jimpy mice. Mol Cell Biol 6:3755–3762
Garin-Chesa P, Rettig WJ, Thomson TM, Old LJ, Melamed MR (1988) Immunohistochemical analysis of nerve growth factor receptor expression in normal and malignant human tissues. J Histochem Cytochem 36:383–389
Hara S, Adachi Y, Kaneko Y, Fujimoto J, Hata J (1991) Evidence for heterogenous groups of neuronal differentiation of Swing's sarcoma. Br J Cancer 64:1025–1030
Hara S, Ishii E, Fujimoto J, Hata J (1989) A monoclonal antibody specifically reactive with Swing's sarcoma. Br J Cancer 60:875–879
Hasegawa T, Hirose T, Kudo E, Hizawa K, Yamawaki S, Ishii S (1991) Atypical primitive neuroectodermal tumors. Comparative light and electron microscopic and immunohistochemical studies on peripheral neuroepitheliomas and Swing's sarcomas. Acta Pathol Jpn 41:444–454
Hempstead BL, Schleifer LS, Chao M (1989) Expression of functional nerve growth factor receptors after gene transfer. Science 243:373–375
Homma C, Kaneko Y, Sekine K, Hara S, Hata J, Sakurai M (1989) Establishment and characterization of a small round cell sarcoma cell line, SCCH-196, with t(11; 22)(q24; q12). Jpn J Cancer Res 80:861–865
Hu MCT, S.B. S, Davidson N (1986) The complete sequence of the mouse skeletal alpha-actin gene reveals several conserved and inverted repeat sequences outside of the proteincoding region. Mol Cell Biol 6:15–25
Ikenaka K, Nakahira K, Takayama C, Wada K, Hatanaka H, Mikoshiba K (1990) Nerve growth factor rapidly induces expression of the 68-kDa neurofilament gene by posttranscriptional modification in PC12h-R cells. J Biol Chem 265: 19782–19785
Jaffe R, Agostini J, Santamaria M (1984) The neuroectodermal tumour of bone. Am J Surg Pathol 8:885–898
Jeon I-S, Davis JN, Braun BS, Sublett JE, Roussel MF, Denny CT, Shapiro DN (1995) A variant Ewing's sarcoma translocation (7; 22) fuses the EWS gene to the ETS gene ETVl. Oncegene 10:1229–1234
Julien JP, Ramachandran K, Grosveld FG (1985) Cloning of a cDNA encoding the smallest neurofilament protein. Biochim Biophys Acta 825:398–404
Kawaguchi K, Koike M (1986) Neuron-specific enolase and Leu-7 immunoreactive small round-cell neoplasms. The relationship to Ewing's sarcoma in bone and soft tissues. Am J Clin Pathol 86:79–83
Levi-Montalcini R (1987) The nerve growth factor. Thirty-five years later. EMBO J 6:1145–1154
Lewis SA, Cowan NJ (1985) Genetics, evolution, and expression of the 68,000 mol-wt neurofilament protein. Isolation of a cloned cDNA probe. J Cell Biol 100:843–850
Lipinski M, Braham K, Philip I, Wiels J, Philip T, Goridis C, Lenoir GM, Tursz T (1987) Neuroectoderm-associated antigens on Swing's sarcoma cell lines. Cancer Res 47:183–187
Lizard-Nacol S, Lizard G, Justrabo E, Turc-Carel C (1989) Immunologic characterization of Swing's sarcoma using mesenchymal and neural markers. Am J Pathol 135:847–855
Llombart-Bosch A, Lacombe MJ, Contesso G, Peydro-Olaya A (1987) Small round blue cell sarcoma of bone mimicking atypical Swing's sarcoma with neuroectodermal features. An analysis of five cases with immunohistochemical and electron microscopic support. Cancer 60:1570–1582
Llombart-Bosch A, Lacombe MJ, Peydro-Olaya A, Perz-Bacete M, Contesso G (1988) Malignant peripheral neuroectodermal tumours of bone other than Askin's neoplasm: characterization of 14 new cases with immunohistochemistry and electron miroscope. Virchows Arch [A] 412:421–430
Marina NM, Etcubanas E, Parham DM, Bowman LC, Green A (1989) Peripheral primitive neuroectodermal tumour (peripheral neuroepitehlioma) in children. A review of the St. Jude experience and controversies in diagnosis and management. Cancer 64:1952–1960
Mierau G (1985) Extraskeletal Swing's sarcoma (peripheral neuroepithelioma). Ultrastruct Pathol 9:91–98
Misko TP, Radeke MJ, Shooter EM (1988) Neural development and regeneration. Springer-Verlag, Berlin, pp 257–272
Myers MW, Lazzarini RA, Lee VM-Y, Schaepfer WW, Nelson DL (1987) The human mid-size neurofilament subunit. A repeated protein sequence and the relationship of its gene to the interfilament gene family. EMBO J 6:1617–1626
Nakakihara K, Ikenaka K, Wada K, Tamura T, Furuichi T, Mikoshiba K (1990) Structure of the 68-kDa neurofilament gene and regulation of its expression. J Biol Chem 265:19786–19791
Noguera R, Navarro S, Peydro OA, Llombart BA (1994) Patterns of differentiation in extraosseous Swing's sarcoma cells. An in vitro study. Cancer 73:616–24
Noguera R, Triche TJ, Navarro S, Tsokos M, Llombart BA (1992) Dynamic model of differentiation in Swing's sarcoma cells. Comparative analysis of morphologic, immunocytochemical, and oncogene expression parameters. Lab Invest 66: 143–151
Okuda T, Kasai K, Kameya T, Saito S, Takase N (1988) Monoclonal antibody directed against neuroendocrine properties of both normal and malignant cells. Hybridoma 7:569–581
Plougastel B, Zucman J, Peter M, Thomas G, Delattre O (1993) Genomic structure of the EWS gene and its relationship to EWSR1, a site of tumour-associated chromosome translocation. Genomics 18:609–615
Rao VN, Papas TS, Reddy SP (1987) ERG, A human ets-related gene on chromosome 21: alternative splicing, polyadenylation, and translation. Science 237:635–639
Schlaepfer WW, Bruce J (1990) Simultaneous up-regulation of neurofilament proteins during the postnatal development of the rat nervous system. J Neurosci Res 25:39–49
Schmidt D, Herrmann C, Jurgens H, Harms D (1991) Malignant peripheral neuroectodermal tumour and its necessary distinction from Ewing's sarcoma. A report from the Kiel Pediatric Tumor Registry. Cancer 68:2251–2259
Shaw G, Weber K (1982) Differential expression of neurofilament triplet proteins in brain developmen. Nature 298:277–279
Shimada H, Newton WA, Soule EH, Qualman SJ, Aoyama C, Mauer HM (1988) Pathologic features of extraosseous Swing's sarcoma: a report from the Intergroup Rhabdomyosarcoma Study. Human Pathol 19:442–453
Sorensen PHB, Lessnick SL, Lopez-Terrada D, Liu XF, Triche TJ, Denny CT (1994) A second Swing's sarcoma transocation, t(21;22), fuses the EWS gene to another STS-family transcription factor, ERG. Nature Genetics 6:146–151
Steinert PM, Jones JCR, Goldman RD (1984) Intermediate filament. J Cell Biol 99:22–27
Trojanovski JQ, Walkenstein N, Lee VM-Y (1986) Expression of neurofilament subunits in neurons of the central and peripheral nervous system. An immunohistochemical study with monoclonal antibodies. J Neurosci 6:650–660
Turc-Carel C, Philip I, Berger MP, Lemoir GM (1983) Chromosomal translocation Swing's sarcoma. New Engl J Med 309:497–498
Umezawa A, Hata J (1992) Expression of gap-junctional protein (connexin 43 or alpha 1 gap junction) is down-regulated at the transcriptional level during adipocyte differentiation of H1/A marrow stromal cells. Cell Struct Funct 17:177–184
Umezawa A, Tachibana K, Harigaya K, Kusakari S, Kato S, Watanabe Y, Takano T (1991) Colony-stimulating factor 1 expression is down-regulated during the adipocyte differentiation of H-1/A marrow stromal cells and induced by cachectin/tumor necrosis factor. Mol Cell Biol 11:920–927
Urano F, Umezawa A, Hong W, Kikuchi H, Hata J-i (1996) A novel chimera gene between EWS and EIA-F, encoding the adenovirus EIA enhancer-binding protein, in extraosseous Swing's sarcoma. Biochem Biophy Res Comm 219:608–612
Ushigome S, Shimoda T, Nikaido T, Nakamori K, Miyazawa Y, Shishikura A, Takakuwa T, Ubayama Y, Spjut HJ (1992) Primitive neuroectodermal tumours of bone and soft tissue. With reference to histologic differentiation in primary or metastatic foci. Acta Pathol Jpn 42:483–493
Whang-Peng J, Triche TJ, Knusten T, Miser J, Douglas EC, Israel MA (1984) Chromosomal translocation in peripheral neuroepithelioma. New Engl J Med 311:584–585
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Sugimoto, T., Umezawa, A. & Hata, Ji. Neurogenic potential of Ewing's sarcoma cells. Virchows Archiv 430, 41–46 (1997). https://doi.org/10.1007/BF01008015
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DOI: https://doi.org/10.1007/BF01008015