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Sci Rep. 2018 Apr 13;8(1):5949. doi: 10.1038/s41598-018-24503-y.

hTERT gene knockdown enhances response to radio- and chemotherapy in head and neck cancer cell lines through a DNA damage pathway modification.

Author information

1
Department of Head and Neck Surgery, Poznan University of Medical Sciences, The Greater Poland Cancer Centre, Garbary 15 Str., 61-866, Poznan, Poland. wojciech.barczak@wco.pl.
2
Radiobiology Lab, The Greater Poland Cancer Centre, Garbary 15 Str., 61-866, Poznan, Poland. wojciech.barczak@wco.pl.
3
Department of Head and Neck Surgery, Poznan University of Medical Sciences, The Greater Poland Cancer Centre, Garbary 15 Str., 61-866, Poznan, Poland.
4
Radiobiology Lab, The Greater Poland Cancer Centre, Garbary 15 Str., 61-866, Poznan, Poland.
5
Head and Neck Cancer Biology Lab, Department of Biology and Environmental Studies, Poznan University of Medical Sciences, Poznan, Poland.
6
University of Central Florida, Burnett School of Biomedical Sciences, College of Medicine, FL, 32827, Orlando, USA.
7
Department of Clinical Chemistry and Molecular Diagnostics, Poznan University of Medical Sciences, Przybyszewskiego 49 Str., 60-355, Poznan, Poland.
8
Department of Electroradiology, Poznan University of Medical Sciences, Garbary 15 Str., 61-866, Poznan, Poland.

Abstract

The aim of the study was to analyze the effect of hTERT gene knockdown in HNSCC cells by using novel in vitro models of head and neck cancer (HNSCC), as well as improving its personalized therapy. To obtain the most efficient knockdown siRNA, shRNA-bearing lentiviral vectors were used. The efficiency of hTERT silencing was verified with qPCR, Western blot, and immunofluorescence staining. Subsequently, the type of cell death and DNA repair mechanism induction after hTERT knockdown was assessed with the same methods, followed by flow cytometry. The effect of a combined treatment with hTERT gene knockdown on Double-Strand Breaks levels was also evaluated by flow cytometry. Results showed that the designed siRNAs and shRNAs were effective in hTERT knockdown in HNSCC cells. Depending on a cell line, hTERT knockdown led to a cell cycle arrest either in phase G1 or phase S/G2. Induction of apoptosis after hTERT downregulation with siRNA was observed. Additionally, hTERT targeting with lentiviruses, followed by cytostatics administration, led to induction of apoptosis. Interestingly, an increase in Double-Strand Breaks accompanied by activation of the main DNA repair mechanism, NER, was also observed. Altogether, we conclude that hTERT knockdown significantly contributes to the efficacy of HNSCC treatment.

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