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Zebrafish xenotransplantation model for cancer stem-like cell study and high-throughput screening of inhibitors

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Tumor Biology

Abstract

Xenotransplantation studies are important tools for studying cancer biology, especially for assaying tumor cell malignancy and providing cancer information in vivo. Cancer stem-like cells (CSCs) have been identified in many cancer types to drive tumor growth and recurrence, from "keeping" to "keep" resistant to chemotherapy and radiation therapy. In this study, we developed the xenotransplantation of CSCs derived from the leukemia and solid tumor cell lines using the zebrafish models. In adult zebrafish, we investigated that the xenografted leukemia stem cells (LSCs) from K562 cells could proliferate in vivo and keep the cancer property by re-transplantation. As for the solid tumor, these CSCs from DU145 cells (human prostate cancer) and HepG2 cells (human liver cancer) could form the tumor mass and even metastasis after xenotransplantation. In addition, the zebrafish embryos with CSC xenotransplantation could evaluate docetaxel in vivo efficiently and be available to screen the novel inhibitors by high-throughput manner. In summary, these zebrafish xenotransplantation models devote a good platform for the CSC mechanism investigation and anti-CSC inhibitor screening.

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Acknowledgments

We thank M. Ariyoshi and S. Ichikawa for their assistance in the experiments and breeding the fish and R. Ikeyama and Y. Tamura for the secretarial assistance.

Conflicts of interest

None

Funding sources

This work was supported in part by the Japan Science and Technology Agency, KAKENHI of Grants-in-Aid for Scientific Research, and the New Energy and Industrial Technology Development Organization.

Author contribution

T.T. and Y.S. conceived and designed the study. B.Z., J.K., Y.S., N.U., and Y.N. performed the assays and analyzed the data. T.N, T.S., and T.M. performed chemical synthesis assays. B.Z. wrote the paper.

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Authors and Affiliations

  1. Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharmacoinformatics, Mie University Graduate School of Medicine, Mie, Japan

    Beibei Zhang, Yasuhito Shimada, Junya Kuroyanagi, Yuhei Nishimura, Noriko Umemoto & Toshio Tanaka

  2. Mie University Medical Zebrafish Research Center, Mie, Japan

    Yasuhito Shimada, Yuhei Nishimura & Toshio Tanaka

  3. Department of Bioinformatics, Mie University Life Science Research Center, Mie, Japan

    Yasuhito Shimada, Yuhei Nishimura & Toshio Tanaka

  4. Department of Omics Medicine, Mie University Industrial Technology Innovation Institute, Mie, Japan

    Yasuhito Shimada, Yuhei Nishimura & Toshio Tanaka

  5. Corporate R&D Headquarters, Canon Inc, Ohta-ku, Tokyo, 146-8501, Japan

    Tsuyoshi Nomoto, Taichi Shintou & Takeshi Miyazaki

Authors
  1. Beibei Zhang
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  2. Yasuhito Shimada
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  3. Junya Kuroyanagi
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  4. Yuhei Nishimura
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  9. Toshio Tanaka
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Corresponding author

Correspondence to Toshio Tanaka.

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Fig. S1

Different cancer processes of LSC and NC xenografts. (a) K562 cells expressed Kusabira-Orange (KOr) fluorescent protein after transfection. (b) KOr positive cells (indicated by yellow arrows) in the blood from LSC and NC xenotransplanted zebrafish at 35 dpi. (c) The agarose gel electrophoresis for KOr gene in the blood of xenografted fish (35 dpi). C, Control fish without cancer injection, L, LSCs injected zebrafish, N, NCs injected zebrafish, M, Marker. (d) The agarose gel electrophoresis for kor gene after 10 days culture of the KOr positive cells after isolation from fish blood. (PDF 247 kb)

Fig. S2

CSC proliferations were inhibited by chemicals in vitro. (a) Docetaxel inhibited ALDH + cells of DU145-KOr after 24 h exposure at different doses. n = 4, **P < 0.01. (b) One fluorescent chemical B647 inhibited ALDH + cells of K562-BFP after 24 h exposure at different doses. n = 4, **P < 0.01. (PDF 166 kb)

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Zhang, B., Shimada, Y., Kuroyanagi, J. et al. Zebrafish xenotransplantation model for cancer stem-like cell study and high-throughput screening of inhibitors. Tumor Biol. 35, 11861–11869 (2014). https://doi.org/10.1007/s13277-014-2417-8

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  • DOI: https://doi.org/10.1007/s13277-014-2417-8

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