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Cancer stem cell markers in pediatric sarcomas: Sox2 is associated with tumorigenicity in immunodeficient mice

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Tumor Biology

Abstract

The three most frequent pediatric sarcomas, i.e., Ewing’s sarcoma, osteosarcoma, and rhabdomyosarcoma, were examined in this study: three cell lines derived from three primary tumor samples were analyzed from each of these tumor types. Detailed comparative analysis of the expression of three putative cancer stem cell markers related to sarcomas—ABCG2, CD133, and nestin—was performed on both primary tumor tissues and corresponding cell lines. The obtained results showed that the frequency of ABCG2-positive and CD133-positive cells was predominantly increased in the respective cell lines but that the high levels of nestin expression were reduced in both osteosarcomas and rhabdomyosarcomas under in vitro conditions. These findings suggest the selection advantage of cells expressing ABCG2 or CD133, but the functional tests in NOD/SCID gamma mice did not confirm the tumorigenic potential of cells harboring this phenotype. Subsequent analysis of the expression of common stem cell markers revealed an evident relationship between the expression of the transcription factor Sox2 and the tumorigenicity of the cell lines in immunodeficient mice: the Sox2 levels were highest in the two cell lines that were demonstrated as tumorigenic. Furthermore, Sox2-positive cells were found in the respective primary tumors and all xenograft tumors showed apparent accumulation of these cells. All of these findings support our conclusion that regardless of the expression of ABCG2, CD133 and nestin, only cells displaying increased Sox2 expression are directly involved in tumor initiation and growth; therefore, these cells fit the definition of the cancer stem cell phenotype.

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Acknowledgments

The authors thank Johana Maresova, Marcela Vesela, and Dr. Jan Verner for their skillful technical assistance. This study was supported by the project no. NT13443-4 from the Internal Grant Agency of the Czech Ministry of Healthcare, by the project no. LQ1605 from the National Program of Sustainability II, and by the European Regional Development Fund—Project CEB no. CZ.1.07/2.3.00/20.0183.

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Authors and Affiliations

  1. Department of Experimental Biology, School of Science, Masaryk University, Brno, Czech Republic

    Jan Skoda, Alena Nunukova, Tomas Loja, Jakub Neradil & Renata Veselska

  2. Department of Pediatric Oncology, University Hospital Brno and School of Medicine, Masaryk University, Brno, Czech Republic

    Jan Skoda, Jakub Neradil, Peter Mudry, Karel Zitterbart, Jaroslav Sterba & Renata Veselska

  3. International Clinical Research Center, St. Anne’s University Hospital Brno, Brno, Czech Republic

    Jan Skoda & Renata Veselska

  4. 1st Institute of Pathologic Anatomy, St. Anne’s University Hospital and School of Medicine, Masaryk University, Brno, Czech Republic

    Iva Zambo & Marketa Hermanova

  5. Department of Histology and Embryology, School of Medicine, Masaryk University, Brno, Czech Republic

    Ales Hampl

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  1. Jan Skoda
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  2. Alena Nunukova
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Correspondence to Renata Veselska.

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The Research Ethics Committee of the School of Science (Masaryk University) approved the study protocol, and a written statement of informed consent was obtained from each participant or his/her legal guardian prior to participation in this study. This study was approved by the Institutional Animal Care and Use Committee of Masaryk University and was registered by the Ministry of Agriculture of the Czech Republic as required by national legislation.

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Skoda, J., Nunukova, A., Loja, T. et al. Cancer stem cell markers in pediatric sarcomas: Sox2 is associated with tumorigenicity in immunodeficient mice. Tumor Biol. 37, 9535–9548 (2016). https://doi.org/10.1007/s13277-016-4837-0

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