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Establishment and proteomic characterization of patient-derived clear cell sarcoma xenografts and cell lines

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Abstract

Clear cell sarcoma (CCS) is an aggressive mesenchymal malignancy characterized by the unique chimeric EWS-ATF1 fusion gene. Patient-derived cancer models are essential tools for the understanding of tumorigenesis and the development of anti-cancer drugs; however, only a limited number of CCS cell lines exist. The objective of this study was to establish patient-derived CCS models. We established patient-derived CCS models from a 43-yr-old female patient. We prepared the patient-derived xenografts (PDXs) from tumor tissues obtained through biopsy or surgery and isolated stable cell lines from PDXs and the original tumor tissue. The presence of gene fusions was examined by RT-PCR, and Sanger sequencing. The established cell lines were characterized by short tandem repeat, viability, colony and spheroid formation, and invasion analyses. Differences in gene enrichment between the primary tumor and cell lines were examined by mass spectrometry and KEGG pathway analysis. The cell lines were maintained for more than 80 passages, and had tumorigenic characteristics such as colony and spheroid formation and invasion. Mass spectrometric proteome analysis demonstrated that the cell lines were enriched for similar but distinct molecular pathways, compared to those in the xenografts and original tumor tissue. Next, tyrosine kinase inhibitors were screened for their suppressive effects on viability. We found that ponatinib, vandetanib, and doxorubicin suppressed the growth of cell lines, and had equivalent IC50 values. Further in-depth investigation and understanding of drug-sensitivity mechanisms will be important for the clinical applications of our cell lines.

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Abbreviations

DAVID:

Database for Annotation, Visualization and Integration Discovery

KEGG:

Kyoto Encyclopedia of Genes and Genomes

LC-MS/MS:

liquid-chromatography combined mass spectrometry

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Acknowledgments

We thank Drs. Y. Minami, K. Shimizu, T. Mori, T. Uehara, M. Sugawara, Y. Araki, S. Toki, and Ms. R. Nakano (Division of Musculoskeletal Oncology, National Cancer Center Hospital) for sampling tumor tissue specimens from surgically-resected materials. We would like to thank Editage (www.editage.jp) for English language editing and constructive comments regarding the manuscript.

Funding

This work was supported by the National Cancer Center Research and Development Fund [26-A-3, 26-A-9, and 29-A-2].

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Authors and Affiliations

  1. Department of Innovative Seeds Evaluation, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku, Tokyo, 104-0045, Japan

    Marimu Sakumoto, Rieko Oyama, Yoko Takai, Fusako Kito & Tadashi Kondo

  2. Central Animal Division, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku, Tokyo, 104-0045, Japan

    Mami Takahashi

  3. Division of Rare Cancer Research, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku, Tokyo, 104-0045, Japan

    Kumiko Shiozawa, Zhiwei Qiao & Tadashi Kondo

  4. Division of Musculoskeletal Oncology, National Cancer Center Hospital, 5-1-1 Tsukiji, Chuo-ku, Tokyo, 104-0045, Japan

    Makoto Endo & Akira Kawai

  5. Department of Pathology and Clinical Laboratories, National Cancer Center Hospital, 5-1-1 Tsukiji, Chuo-ku, Tokyo, 104-0045, Japan

    Akihiko Yoshida

Authors
  1. Marimu Sakumoto
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  2. Rieko Oyama
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Correspondence to Tadashi Kondo.

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Editor: Tetsuji Okamoto

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Sakumoto, M., Oyama, R., Takahashi, M. et al. Establishment and proteomic characterization of patient-derived clear cell sarcoma xenografts and cell lines. In Vitro Cell.Dev.Biol.-Animal 54, 163–176 (2018). https://doi.org/10.1007/s11626-017-0207-5

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