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Isolation of tumor stem-like cells from primary laryngeal squamous cell carcinoma cells (FD-LS-6)

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Abstract

The development of efficient treatments for laryngeal squamous cell carcinoma (LSCC) is hindered by the lack of applicable tumor cell lines and animal models of the disease, especially those related to cancer stem-like cells (CSCs). CSCs play critical roles in tumor propagation and pathogenesis whereas no CSCs lines have been developed to date. In this study, we establish an LSCC cell line (FD-LS-6) from primary LSCC tumor tissue (not experienced single-cell cloning) and adapted a culturing condition for the expansion of potential stem cells (EPSCs) to isolate CSCs from FD-LS-6. We successfully derived novel CSCs and named them as LSCC sphere-forming cells (LSCSCs) which were subsequently characterized for their CSC properties. We showed that LSCSCs shared many properties of CSCs, including CSC marker, robust self-renewal capacity, tumorigenesis ability, potential to generate other cell types such as adipocytes and osteoblasts, and resistance to chemotherapy. Compared to parental cells, LSCSCs were significantly more potent in forming tumors in vivo in mice and more resistant to chemotherapy. LSCSCs have higher expressions of epithelial–mesenchymal transition proteins and chemotherapy resistance factors, and exhibit an activated COX2/PEG2 signaling pathway. Altogether, our work establishes the first CSCs of LSCC (FD-LS-6) and provides a tool to study tumorigenesis and metastasis of LSCC and help the development of anticancer therapies.

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Abbreviations

LSCC:

Laryngeal squamous cell carcinoma

HNSCC:

Head and neck squamous cell carcinoma

CSCs:

Cancer stem-like cells

EPSC:

Expanded potential stem cells

LSCSCs:

LSCC sphere-forming cells

PBS:

Phosphate buffer saline

SP:

Side population

BEGM:

Bronchial epithelial cell growth medium

EMT:

Epithelial–mesenchymal transition

MET:

Mesenchymal- epithelial transition

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Funding

This study was supported by the National Natural Science Foundation of China under Grand (82103316, 82203506, 30801283, 30972691, 82071856, 81671579, 31370904). Program for scientific and technological innovation from the Science and Technology Commission of Shanghai Municipality (22490760400); Shanghai Municipal Commission of Health, Scientific Research Program of Traditional Chinese medicine(2020JP009); Shuguang Planning of Shanghai Municipal Education Commission (16SG14); The National Key Research and Development Program (2020YFA0113101).

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Author notes

  1. Duo Zhang and Di Tang have contributed equally to this work.

Authors and Affiliations

  1. Shanghai Institute of Immunology, Shanghai Jiao Tong University School of Medicine, 280 South Chongqing Road, Shanghai, 200025, China

    Li-ming Lu

  2. Department of Otolaryngology-HNS, Eye, Ear, Nose and Throat Hospital, Shanghai Key Clinical Disciplines of Otorhinolaryngology, Fudan University School of Medicine, 83 Fenyang Road, Shanghai, 200031, China

    Duo Zhang, Di Tang & Lei Tao

  3. Department of Pudong Hospital, Fudan University School of Medicine, 2800 Gongwei Road, Shanghai, 201300, China

    Duo Zhang, Di Tang & Lei Tao

  4. School of Biomedical Sciences, Stem Cell and Regenerative Medicine Consortium, Li Ka Shing Faculty of Medicine, The University of Hong Kong, 5 Sassoon Road, Hong Kong, China

    Pen-tao Liu

  5. Centre for Translational Stem Cell Biology, Science and Technology Park, 6-8 Harbour Road, Hong Kong, China

    Pen-tao Liu

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  1. Duo Zhang
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Correspondence to Lei Tao or Li-ming Lu.

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Ethical approval

This study was approved by ethics boards of the Eye and ENT Hospital of Fudan University and was conducted in line with the principles of the Declaration of Helsinki (No. KJ2008-01).

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Supplementary Information

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13577_2023_984_MOESM1_ESM.jpg

Supplementary file1 (JPG 1327 KB) Short tandem repeat (STR) analysis showed the primary cultured LSCCs were human-derived cells without cross-contamination by other cell lines. (A) D3S1358, TH01, D21S11, D18S51 and Penta E were labelled by FL. (B) D13S317, D16S539, CSF1PO, D7S820, D5S818and Penta D were labelled by JOE. (C) Amelogenin, FGA, TPOX, vWA and D8S1179 were labelled by TMR. (D) D19S433 and D2S1338 were labelled by CXR

Supplementary file2 (TIF 8111 KB) AMC-HN-8 in EPSCM

Supplementary file3 (XLSX 13 KB)

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Zhang, D., Tang, D., Liu, Pt. et al. Isolation of tumor stem-like cells from primary laryngeal squamous cell carcinoma cells (FD-LS-6). Human Cell 37, 323–336 (2024). https://doi.org/10.1007/s13577-023-00984-6

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  • DOI: https://doi.org/10.1007/s13577-023-00984-6

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