Identification, expansion and characterization of cancer cells with stem cell properties from head and neck squamous cell carcinomas

• Published in: Experimental cell research Vol. 348; no. 1; pp. 75 - 86
• Main Authors: Kaseb, Hatem O., Fohrer-Ting, Helene, Lewis, Dale W., Lagasse, Eric, Gollin, Susanne M.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 15.10.2016; Elsevier BV; Elsevier
• Subjects: 60 APPLIED LIFE SCIENCES; Adaptive immunology; Aged; Aged, 80 and over; Animals; Antigens, CD - metabolism; BIOLOGICAL MARKERS; Biomarkers, Tumor - metabolism; Cancer; Cancer stem cells; Carcinogenesis - pathology; Carcinoma, Squamous Cell - metabolism; Carcinoma, Squamous Cell - pathology; CARCINOMAS; CELL CULTURES; Cell Line, Tumor; Cell Proliferation; Cell Self Renewal; Cell Separation; Cell surface markers; Cellular biology; Chromosomal Instability; CHROMOSOMES; Clone Cells; COMPARATIVE EVALUATIONS; DILUTION; EFFICIENCY; Feeder Cells - cytology; Female; FLUORESCENCE; Fluorescent Antibody Technique; HEAD; Head & neck cancer; Head and Neck Neoplasms - metabolism; Head and Neck Neoplasms - pathology; Head and neck squamous cell carcinoma; Humans; Immunology; In Situ Hybridization, Fluorescence; IN VITRO; IN VIVO; Innate immunity; INSTABILITY; IONIZING RADIATIONS; IRRADIATION; Life Sciences; LUNGS; Male; MICE; Middle Aged; NECK; Neoplastic Stem Cells - metabolism; Neoplastic Stem Cells - pathology; PATIENTS; PUBLIC HEALTH; Radiation Tolerance; Radioresistance; RADIOSENSITIVITY; SOLIDS; SPHEROIDS; Squamous Cell Carcinoma of Head and Neck; STEM CELLS; TUMOR CELLS; CFE; ELDA; NSCLC; OS; Cell surface markers; CIN; IR; Radioresistance; Chromosomal instability; SC; CSC; HNSCC; FISH; Head and neck squamous cell carcinoma; Cancer stem cells; SP
• ISSN: 0014-4827; 1090-2422
• DOI: 10.1016/j.yexcr.2016.09.003

Head and neck squamous cell carcinoma (HNSCC) is a major public health concern. Recent data indicate the presence of cancer stem cells (CSC) in many solid tumors, including HNSCC. Here, we assessed the stem cell (SC) characteristics, including cell surface markers, radioresistance, chromosomal instability, and in vivo tumorigenic capacity of CSC isolated from HNSCC patient specimens. We show that spheroid enrichment of CSC from early and short-term HNSCC cell cultures was associated with increased expression of CD44, CD133, SOX2 and BMI1 compared with normal oral epithelial cells. On immunophenotyping, five of 12 SC/CSC markers were homogenously expressed in all tumor cultures, while one of 12 was negative, four of 12 showed variable expression, and two of the 12 were expressed heterogeneously. We showed that irradiated CSCs survived and retained their self-renewal capacity across different ionizing radiation (IR) regimens. Fluorescence in situ hybridization (FISH) analyses of parental and clonally-derived tumor cells revealed different chromosome copy numbers from cell to cell, suggesting the presence of chromosomal instability in HNSCC CSC. Further, our in vitro and in vivo mouse engraftment studies suggest that CD44+/CD66− is a promising, consistent biomarker combination for HNSCC CSC. Overall, our findings add further evidence to the proposed role of HNSCC CSCs in therapeutic resistance. •Spheroid enrichment selects cancer stem cells (CSC) from head & neck tumors (HNSCC).•Compared to normal epithelial cells, isolated CSC express increased SC/CSC markers.•Isolated CSC display enhanced radioresistance, clonogenicity and tumorigenicity.•HNSCC CSC express chromosomal instability.•CD44+/CD66− is a promising, consistent biomarker for HNSCC CSC.