A novel human in vitro papillomavirus type 16 positive tonsil cancer cell line with high sensitivity to radiation and cisplatin

BMC Cancer. 2019 Mar 25;19(1):265. doi: 10.1186/s12885-019-5469-8.

Abstract

Background: Human papillomavirus (HPV) is an established risk factor for oropharyngeal squamous cell carcinoma (OSCC). The aim was to establish cell lines from HPV-positive tonsil carcinomas to be used for treatment development.

Methods: Fresh samples from 23 HPV-positive tonsil carcinomas were cultivated in vitro. The established cell line was analyzed for viral characteristics, cell karyotype, TP53 status, and growth capabilities in nude mice. In vitro studies of sensitivities to radiation, cisplatin and cetuximab were performed.

Results: After 19 months (eight passages), one cell line, LU-HNSCC-26, was established in vitro and also grew as xenografts. The tumor was from a 48 year old non-smoking man with non-keratinizing, p16 positive tonsil OSCC, stage T2N0M0 with HPV16. It contained 19.5 (CV% 3.7) HPV16 copies/cell (passage 8). The complete HPV16 genome sequence was obtained. Episomal HPV16 was present with an E2/E7 ratio of 1.1 (CV% 2.6). In addition, HPV16 mRNA specific for the intact E2 gene was detected. The viral expression manifested 1.0 (CV% 0.1) E7 mRNA copies per HPV16 genome. The karyotype was determined and the cell line demonstrated wild type TP53. The ID50 for radiation was 0.90 Gy and the IC50 for cisplatin was 0.99 μmol/L. The cell line was inhibited to a maximum of 18% by cetuximab.

Conclusions: We established an in vitro tonsil carcinoma cell line containing episomal HPV16. This is an important step towards efficient treatment development.

Keywords: Cell line; Cetuximab; Cisplatin; Human papilloma virus; Radiation; Tonsil cancer.

MeSH terms

  • Animals
  • Cell Culture Techniques / methods*
  • Cell Line, Tumor / cytology*
  • Cell Line, Tumor / virology
  • Cell Survival / drug effects
  • Cell Survival / radiation effects
  • Cisplatin / administration & dosage*
  • Cisplatin / therapeutic use
  • Genome, Viral
  • Human papillomavirus 16 / drug effects
  • Human papillomavirus 16 / genetics*
  • Human papillomavirus 16 / radiation effects
  • Humans
  • Inhibitory Concentration 50
  • Karyotype
  • Male
  • Mice
  • Middle Aged
  • Papillomavirus Infections / genetics
  • Papillomavirus Infections / therapy*
  • Papillomavirus Infections / virology
  • Radiotherapy
  • Tonsillar Neoplasms / genetics
  • Tonsillar Neoplasms / therapy
  • Tonsillar Neoplasms / virology*
  • Viral Load / drug effects
  • Viral Load / radiation effects
  • Whole Genome Sequencing
  • Xenograft Model Antitumor Assays

Substances

  • Cisplatin