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

Abstract

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.

Inducible Expression of Chimeric EWS/ETS Proteins Confers Ewing's Family Tumor-Like Phenotypes to Human Mesenchymal Progenitor Cells

“…UET-13 cells were cultured in Dulbecco's modified Eagle's medium (DMEM) with 10% Tet system approved fetal bovine serum (T-FBS) (Takara) at 37°C under a humidified 5% CO 2 atmosphere. EFT cell lines (EES-1 [20], SCCH196 [21], RD-ES and SK-ES1 [5], NCR-EW2 and NCR-EW3 [19], and W-ES [13]) and neuroblastoma (NB) cell lines (NB69 and NB9 [15] and GOTO [47]) were cultured in RPMI 1640 with 10% FBS. A rhabdomyosarcoma cell line, NRS-1 (40), was cultured in Eagle's minimal essential medium with 10% FBS.…”

Extraosseous Ewing's sarcoma arising from the pterygomandibular space

“…Both EES and pPNET show varying degree of neuroectodermal differentiation [11]. The term EES has been used for those tumors that lack neuroectodermal differentiation by light microscopy, immunohistochemistry and electron microscopy [18]. pPNET shows neuroectodermal differentiation, with rosette formation and immunohistochemical staining for nucleus including neuron specific enolase (NSE), neurofilament (NF) and synaptophysin (Syn) .Currently they are categorized into a group known as the Ewing sarcoma family of tumors (ESFT) which includes classic ES of bone, EES, Askin tumors of the chest wall and pPNET of bone or soft tissues.…”

The establishment and characterization of a continuous cell line of mouse cervical carcinoma

“…The transfection was performed according to the instructions of the Lipofectamine 2000 supplier (Invitrogen). After incubation of the cells for 24 h at 37 under 5% CO 2 , the efficiency of transfection DMI 4000B) [8] . After 48 h, the cells were transfered to selective media containing G418 sulfate (800 μg/ml).…”