Sensitivity of human papillomavirus‑positive and ‑negative oropharyngeal cancer cell lines to ionizing irradiation

Holzhauser,Stefan; Pirotte,Evelyne; Jones,Joanne; Owens,David; Al‑Hussaini,Ali; Giles,Peter; Evans,Mererid; Man,Stephen; Powell,Ned

doi:10.3892/or.2020.7709

Sensitivity of human papillomavirus‑positive and ‑negative oropharyngeal cancer cell lines to ionizing irradiation

Authors:
- Stefan Holzhauser
- Evelyne Pirotte
- Joanne Jones
- David Owens
- Ali Al‑Hussaini
- Peter Giles
- Mererid Evans
- Stephen Man
- Ned Powell
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Affiliations: HPV Research Group, Cardiff University, School of Medicine, Cardiff CF14 4XN, UK, Ear Nose/Throat/Head and Neck Surgery Department, University Hospital of Wales, Cardiff and Vale University Health Board, Cardiff CF14 4XW, UK, Wales Gene Park, Cardiff University, Cardiff CF14 4XN, UK, Velindre Cancer Centre, Cardiff CF14 2TL, UK, Division of Cancer and Genetics, Cardiff University, School of Medicine, Cardiff CF14 4XN, UK
Published online on: July 31, 2020 https://doi.org/10.3892/or.2020.7709
Pages: 1717-1726

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Abstract

Human papillomavirus‑positive (HPV+) oropharyngeal squamous cell carcinoma (OPSCC) has increased in incidence and has a much better prognosis than HPV‑negative (HPV‑) OPSCC with radiotherapy alone, but exactly why is unknown. The present study therefore aimed to further examine the sensitivity and possible changes in gene expression of several HPV+ and HPV‑ OPSCC, including various novel cell lines, upon ionizing irradiation (IR). Previously established HPV+ UM‑SCC‑47, UPCI‑SCC‑90, CU‑OP‑2, CU‑OP‑3 and HPV‑ UM‑SCC‑4, UM‑SCC‑6, UM‑SCC‑74a, UM‑SCC‑19 and newly established CU‑OP‑17 and CU‑OP‑20, characterised here, were subjected to 0‑6 Gy. Surviving fractions of each cell line were tested by clonogenic assays, and irregularities in cell cycle responses were examined by flow cytometry, while changes in gene expression were followed by mRNA sequencing. HPV+ OPSCC cell lines showed greater variation in sensitivity to ionizing irradiation (IR) and tended to be more sensitive than HPV‑ OPSCC cell lines. However, their IR sensitivity was not correlated to the proportion of cells in G2 arrest, and HPV‑ cell lines generally showed lower increases in G2 after IR. Upon IR with 2 Gy, mRNA sequencing revealed an increase in minor HPV integration sites in HPV+ cell lines, and some changes in gene expression in OPSCC cell lines, but not primarily those associated with DNA repair. To conclude, HPV+ OPSCC cell lines showed greater variation in their sensitivity to IR, with some that were radioresistant, but overall the HPV+ OPSCC group still tended to be more sensitive to IR than the HPV‑ OPSCC group. In addition, HPV+ OPSCC lines were more frequently in G2 as compared to HPV‑ cell lines, but the increase in G2 arrest upon IR in HPV+ OPSCC was not correlated to sensitivity to IR. Increases in minor HPV integration sites and changes in gene expression were also demonstrated after irradiation with 2 Gy.

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