In vivo investigation of CD133 as a putative marker of cancer stem cells in Hep-2 cell line

Authors

  • Xu Dong Wei PhD,

    1. Department of Otolaryngology–Head and Neck Surgery, Fudan University, Affiliated Eye, Ear, Nose, and Throat Hospital, Shanghai, China
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  • Liang Zhou MD,

    Corresponding author
    1. Department of Otolaryngology–Head and Neck Surgery, Fudan University, Affiliated Eye, Ear, Nose, and Throat Hospital, Shanghai, China
    • Department of Otolaryngology–Head and Neck Surgery, Fudan University, Affiliated Eye, Ear, Nose, and Throat Hospital, Shanghai, China
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  • Lei Cheng PhD,

    1. Department of Otolaryngology–Head and Neck Surgery, Fudan University, Affiliated Eye, Ear, Nose, and Throat Hospital, Shanghai, China
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  • Jie Tian BM,

    1. Central Laboratory, Affiliated Eye, Ear, Nose and Throat Hospital, Fudan University, Shanghai, China
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  • Jack J. Jiang PhD,

    1. Department of Surgery, Division of Otolaryngology–Head and Neck Surgery, University of Wisconsin Medical School, Madison, Wisconsin
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  • Julia MacCallum PhD

    1. Department of Surgery, Division of Otolaryngology–Head and Neck Surgery, University of Wisconsin Medical School, Madison, Wisconsin
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Abstract

Background

Mounting evidence suggests that most tumors consist of a heterogeneous population of cells with a subset population that has the exclusive tumorigenic ability. They are called cancer stem cells (CSCs). CSCs can self-renew to generate additional CSCs and also differentiate to generate phenotypically diverse cancer cells with limited proliferative potential. They have been identified in a variety of tumors. In this study, we identify the marker of CSCs in the established human laryngeal tumor Hep-2 cell line in vivo. Our in vitro experiment shown as CD133, a 5-transmembrane glycoprotein expressed in Hep-2 cell line. CD133 was supposed as a candidate of CSC in laryngeal carcinoma. In this study, the expression of CD133 was detected in a Hep-2 cell line. Applying the magnetic cell sorting (MACS) technology, we reported the results of purifying CD133 positive cells from a Hep-2 cell line. Three-type cells' tumor-forming ability was examined in vivo to identify the marker of CSCs in Hep-2 cell line.

Methods

CD133 was selected as a putative marker of CSC in laryngeal carcinoma, Hep-2 cell lines. Flow cytometry was used to detect the expression of CD133 in the Hep-2 cell line. Immunomagnetic beads were applied to purify CD133-positive cells. CD133(+), CD133(−) tumor cells, and unsorted Hep-2 cells were injected into severe combined immune deficiency (SCID) mice individually to observe tumor-forming ability.

Results

Only a small proportion (3.15% ± 0.83%) of cells in the Hep-2 cell line express the CD133 marker. In comparison with CD133(−) tumor cells and unsorted cells, CD133(+) cells possess a marked capacity for tumor formation in vivo (p <.05).

Conclusion

CD133 is 1 of the markers for CSCs in human laryngeal tumors of the Hep-2 cell line. Work on the characterization of these cells provides a powerful tool to investigate the tumorigenic process in the larynx and to develop therapies targeting the CSC. © 2008 Wiley Periodicals, Inc. Head Neck, 2009

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