Detection of putative stem cell markers, CD44/CD133, in primary and lymph node metastases in head and neck squamous cell carcinomas. A preliminary immunohistochemical and in vitro study

Clin Otolaryngol. 2015 Aug;40(4):312-20. doi: 10.1111/coa.12368.

Abstract

Objectives: Investigators hypothesized that cancer stem cells (CSCs) could play a role in determining cancer progression by metastasizing to cervical lymph node (N+) and then influencing prognosis of head and neck squamous cell carcinomas (HNSCCs) patients.

Design: To identify CSCs in HNSCCs and their clonogenic capacity.

Setting: In vitro study.

Participants: Putative CSCs from 29 primary HNSCCs and 19 corresponding node metastases were analyzed.

Main outcome measures: Immunohistochemical (IHC) was performed, and CSCs' clonogenic in vitro capacity was tested; ones epithelial nature of cancer cells forming colonies was confirmed by a second IHC, fluorescence-activated cell sorting (FACS) analysis helped in counting CD44/CD133-CSCs markers percentage expression in HNSCC tumour-derived cultures.

Results: Immunohistochemical showed CD44 (93.1%) and CD133 (10.34%) expression; FACS-analysis showed the enrichment of CD44/CD133 cancer cells, with the highest clonogenic capacity of CD44+-subpopulation; a higher CD44 rates were documented from N+ subcultures than from original tumours (P < 0.05).

Conclusions: A putative cancer stem-like cell population is detectable in HNSCCs, and our findings show their in vitro clonogenic capacity by demonstrating that CD44+-cultured cells are the main population proliferating obtained by N+ HNSCC metastases, emphasizing their possible role in tumour progression.

MeSH terms

  • AC133 Antigen
  • Aged
  • Antigens, CD / metabolism*
  • Biomarkers, Tumor / metabolism
  • Carcinoma, Squamous Cell / metabolism*
  • Carcinoma, Squamous Cell / pathology*
  • Female
  • Flow Cytometry
  • Glycoproteins / metabolism*
  • Head and Neck Neoplasms / metabolism*
  • Head and Neck Neoplasms / pathology*
  • Humans
  • Hyaluronan Receptors / metabolism*
  • Immunohistochemistry
  • In Vitro Techniques
  • Lymphatic Metastasis / pathology*
  • Male
  • Middle Aged
  • Neoplastic Stem Cells / metabolism*
  • Peptides / metabolism*
  • Tumor Cells, Cultured

Substances

  • AC133 Antigen
  • Antigens, CD
  • Biomarkers, Tumor
  • Glycoproteins
  • Hyaluronan Receptors
  • PROM1 protein, human
  • Peptides