Johannes Routila1,2,3, Karri Suvila1, Reidar Grénman2, Ilmo Leivo4, Jukka Westermarck1,4, Sami Ventelä5,6,7. 1. Turku Bioscience Centre, University of Turku and Åbo Akademi university, Turku, Finland. 2. Department for Otorhinolaryngology - Head and Neck Surgery, Turku University Hospital and University of Turku, Kiinamyllynkatu 4-8, 20521, Turku, Finland. 3. Department for Otorhinolaryngology, Satakunta Central Hospital, Pori, Finland. 4. Institute of Biomedicine, University of Turku, Kiinamyllynkatu 10, 20520, Turku, Finland. 5. Turku Bioscience Centre, University of Turku and Åbo Akademi university, Turku, Finland. satuve@utu.fi. 6. Department for Otorhinolaryngology - Head and Neck Surgery, Turku University Hospital and University of Turku, Kiinamyllynkatu 4-8, 20521, Turku, Finland. satuve@utu.fi. 7. Institute of Biomedicine, University of Turku, Kiinamyllynkatu 10, 20520, Turku, Finland. satuve@utu.fi.
Abstract
BACKGROUND: Currently, no clinically useful biomarkers for radioresistance are available in head and neck squamous cell carcinoma (HNSCC). This study assesses the usefulness of Cell Line Microarray (CMA) method to enhance immunohistochemical screening of potential immunohistochemical biomarkers for radioresistance in HNSCC cell lines. METHODS: Twenty-nine HNSCC cell lines were cultured, cell pellets formalin-fixed, paraffin-embedded, and arrayed. Radioresistance features of the cell lines were combined to immunohistochemical stains for p53, NDFIP1, EGFR, stem cell marker Oct4, and PP2A inhibitor CIP2A. RESULTS: Expression of p53, EGFR or CIP2A did not indicate intrinsic radioresistance in vitro. Stem cell marker Oct4 nuclear positivity and NDFIP1 nuclear positivity was correlated with increased intrinsic radioresistance. CONCLUSION: The usefulness of CMA in analysis of HNSCC cell lines and discovery of biomarkers is demonstrated. CMA is very well adapted to both testing of antibodies in a large panel of cell lines as well as correlating staining results with other cell line characteristics. In addition, CMA-based antibody screening proved an efficient and relatively simple method to identify potential radioresistance biomarkers in HNSCC.
BACKGROUND: Currently, no clinically useful biomarkers for radioresistance are available in head and neck squamous cell carcinoma (HNSCC). This study assesses the usefulness of Cell Line Microarray (CMA) method to enhance immunohistochemical screening of potential immunohistochemical biomarkers for radioresistance in HNSCC cell lines. METHODS: Twenty-nine HNSCC cell lines were cultured, cell pellets formalin-fixed, paraffin-embedded, and arrayed. Radioresistance features of the cell lines were combined to immunohistochemical stains for p53, NDFIP1, EGFR, stem cell marker Oct4, and PP2A inhibitor CIP2A. RESULTS: Expression of p53, EGFR or CIP2A did not indicate intrinsic radioresistance in vitro. Stem cell marker Oct4 nuclear positivity and NDFIP1 nuclear positivity was correlated with increased intrinsic radioresistance. CONCLUSION: The usefulness of CMA in analysis of HNSCC cell lines and discovery of biomarkers is demonstrated. CMA is very well adapted to both testing of antibodies in a large panel of cell lines as well as correlating staining results with other cell line characteristics. In addition, CMA-based antibody screening proved an efficient and relatively simple method to identify potential radioresistance biomarkers in HNSCC.
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