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KMID : 1001020070050030122
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Korean Journal of Urological Oncology
2007 Volume.5 No. 3 p.122 ~ p.133
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Establishment and Characterization of a Renal Clear Cell Sarcoma Cell Line, CCSK-2
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Lee Young-Goo
Cho Jin-Kuk
Song Ha-Jong
Oh Sook-Kyung
Lee Kwang-Ho
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Abstract
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Purpose: Renal clear cell sarcoma is a rare and highly malignant tumor which is 4-6% of pediatric renal tumor. It had been classified as a variant of Wilms’ tumor. Recently it was separated from the entity of Wilms’ tumor. Cell line is essential to study the characteristics of renal clear cell sarcoma. Here, we report the isolation and characterization of first renal clear cell sarcoma cell line(CCSK-2) from 19-year-old patient.
Materials and Methods : Tumor and normal kidney tissues were harvested from radical nephrectomy specimen, minced with scalpel, plated on tissue culture dishes and maintained under MEM medium with 10% fetal bovine serum. For the characterization, doubling time in vitro, soft agar clonogenic assay, karyotype analysis, immunohistochemistry for vimentin, desmin, smooth muscle actin, S-100 protein, cytokeratin, and reverse transcription- polymerase chain reaction(RT-PCR) for E-cadherin, KAI-1, p53, VHL, WT1, and DCC were performed.
Results: CCSK-2 grows well in vitro(passage number 55) and forms colonies in 0.3% soft agar. Normal kidney cell also grows well under MEM medium. Doubling time of CCSK-2 was 48 hours in vitro. Karyotype showed 49 XY. Immunohistochemistry for vimentin showed strong positive but negative for desmin, smooth muscle actin, S-100 protein, cytokeratin, which supports that CCSK-2 is originated from clear cell sarcoma. RT-PCR showed that E-cadherin, KAI-1, and p53 were not expressed only in CCSK-2, however, VHL, WT1, and DCC were expressed both in normal kidney cell and CCSK-2.
Conclusions : Renal clear cell sarcoma shows very aggressive clinical course and poor prognosis. CCSK-2 is the first renal clear cell sarcoma cell line in the world and grows well in vitro, forms colonies in soft agar, and tumor suppressor genes are not expressed. CCSK-2 will be helpful to study the molecular characteristics of renal clear cell sarcoma and to develop new therapeutic modality for renal clear cell sarcoma.
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KEYWORD
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Renal clear cell sarcoma, Cell line
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