Establishment and characterization of a chordoma cell line from the tissue of a patient with dedifferentiated-type chordoma.
OBJECTIVE Chordoma is a rare bone tumor of the axial skeleton believed to originate from the remnants of the embryonic notochord. The available tumor cells are characteristically physaliferous and express brachyury, a transcription factor critical for mesoderm specification. Although chordomas are histologically not malignant, treatments remain challenging because they are resistant to radiation therapy and because wide resection is impossible in most cases. Therefore, a better understanding of the biology of chordomas using established cell lines may lead to the advancement of effective treatment strategies. The authors undertook a study to obtain this insight. METHODS Chordoma cells were isolated from the tissue of a patient with dedifferentiated-type chordoma (DTC) that had recurred. Cells were cultured with DMEM/F12 containing 10% fetal bovine serum and antibiotics (penicillin and streptomycin). Cell proliferation rate was measured by MTS assay. Cell-cycle distribution and cell surface expression of proteins were analyzed by fluorescence-activated cell sorting (FACS) analysis. Expression of proteins was analyzed by Western blot and immunocytochemistry. Radiation resistance was measured by clonogenic survival assay. Tumor formation was examined by injection of chordoma cells at hindlimb of nude mice. RESULTS The putative (DTC) cells were polygonal and did not have the conventional physaliferous characteristic seen in the U-CH1 cell line. The DTC cells exhibited similar growth rate and cell-cycle distribution, but they exhibited higher clonogenic activity in soft agar than U-CH1 cells. The DTC cells expressed high levels of platelet-derived growth factor receptor-β and a low level of brachyury and cytokeratins; they showed higher expression of stemness-related and epithelial to mesenchymal transition-related proteins than the U-CH1 cells. Intriguingly, FACS analysis revealed that DTC cells exhibited marginal surface expression of CD24 and CD44 and high surface expression of CXCR4 in comparison to U-CH1 cells. In addition, blockade of CXCR4 with its antagonist AMD3100 effectively suppressed the growth of both cell lines. The DTC cells were more resistant to paclitaxel, cisplatin, etoposide, and ionizing radiation than the U-CH1 cells. Injection of DTC cells into the hindlimb region of nude mice resulted in the efficient formation of tumors, and the histology of xenograft tumors was very similar to that of the original patient tumor. CONCLUSIONS The use of the established DTC cells along with preestablished cell lines of chordoma may help bring about greater understanding of the mechanisms underlying the chordoma that will lead to therapeutic strategies targeting chordomas.
Division of Radiation Cancer Research, Research Center for Radio-Senescence.|Department of Pathology, College of Medicine, Korea University, Seoul, South Korea.
Jeong-Yub
JY
Kim
Division of Radiation Cancer Research, Research Center for Radio-Senescence.
Jongsun
J
Lee
Department of Pathology, and.
Jae-Soo
JS
Koh
Division of Radiation Cancer Research, Research Center for Radio-Senescence.
Myung-Jin
MJ
Park
Department of Neurosurgery, Korea Institute of Radiological and Medical Sciences; and.
Ung-Kyu
UK
Chang
D000818:Animals; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects / Q000528:radiation effects; D002817:Chordoma / Q000473:pathology / Q000503:physiopathology / Q000601:surgery; D003050:Coccyx; D005260:Female; D006801:Humans; D008819:Mice, Nude; D008875:Middle Aged; D009364:Neoplasm Recurrence, Local; D009368:Neoplasm Transplantation; D013125:Spinal Neoplasms / Q000473:pathology / Q000503:physiopathology / Q000601:surgery
CXCR4; DTC = dedifferentiated-type chordoma; EGFR = epidermal growth factor receptor; EMT = epithelial to mesenchymal transition; FACS = fluorescence-activated cell sorting; FBS = fetal bovine serum; IR = ionizing radiation; PBS = phosphate-buffered saline; PDGFR = platelet-derived growth factor receptor; PTEN = phosphatase and tensin homolog; RT = room temperature; TBST = Tris-buffered saline with Tween 20; brachyury; chordoma; dedifferentiated-type chordoma; oncology; p- = phosphorylated