Tohoku J. Exp. Med., 2006, 210(3)

Establishment and Characterization of a Clonal Human Extraskeletal Ewing's Sarcoma Cell Line, EES1

MASAHITO HATORI, HIDEYUKI DOI,¹ MIKA WATANABE,² HIRONOBU SASANO,² MASAMI HOSAKA, SATOSHI KOTAJIMA, FUMIHIKO URANO,³ JUNICHI HATA⁴ and SHOICHI KOKUBUN

Department of Orthopaedic Surgery, ¹Division of Advanced Surgical Science and Technology, ²Department of Pathology, Tohoku University School of Medicine, Sendai, Japan, ³Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA, USA, and ⁴National Research Institute for Child Health and Development, Tokyo, Japan

Ewing's sarcoma, a small round cell sarcoma arising in soft tissue as well as the bone, is one of the most malignant tumors in children and young adults. Few established cell lines of extraskeletal Ewing's sarcoma (EES) have been reported, which made it difficult to examine the biological features of EES. Therefore, we have established a new clonal cell line of EES. We report its morphological characters, results of chromosomal and immunohistochemical analysis. A piece of tumor obtained from the 18-year-old female patient with EES was xenografted in a nude mouse. In vitro subcultured cells were then obtained from this xenograft. A clonal cell line was subsequently established by limiting dilution and designated EES1. EES1 cells had a doubling time of 24 hours. In the xenografted tumor, the cells expressed vimentin, CD99 (MIC2), neuron specific enolase (NSE) and cytokeratin. The original tumor cells also expressed vimentin, CD 99, and NSE, but was negative for cytokeratin. The morphological and immunohistochemical features of this cell line established, except for cytokeratin expression, were consistent with those of the primary tumor. Cytogenetic analysis of EES1 revealed chromosomal translocation of t(11; 12)(q24;ql2). The chimeric fusion of the Ewing's sarcoma gene in band 22q12 with the Friend leukemia virus integration-1 gene in band 11q24 was also demonstrated. Fluorescence in situ hybridization further confirmed the presence of translocation involving the Ewing's sarcoma gene in both the primary tumor and EES1 cells. In conclusion, we have established a human EES cell line EES1, which will provide a useful model for studying various aspects of human EES.

keywords —— extraskeletal Ewing's sarcoma; Ewing's sarcoma; cell culture; immunohistochemistry; chromosomal translocation

© 2006 Tohoku University Medical Press

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Tohoku J. Exp. Med., 2006, 210, 221-230

Correspondence: Masahito Hatori, M.D., Department of Orthopaedic Surgery, Tohoku University School of Medicine, 1-1 Seiryo-machi, Aoba-ku, Sendai 980-8574, Japan.

e-mail: mhato@mail.tains.tohoku.ac.jp