Establishment and characterization of an oral tongue squamous cell carcinoma cell line from a never-smoking patient

Steven J. Wang, Saurabh Asthana, Annemieke van Zante, Chase M. Heaton, Janyaporn Phuchareon, Leighton Stein, Saito Higuchi, Tomoya Kishimoto, Charles Y. Chiu, Adam B. Olshen, Frank McCormick, Osamu Tetsu

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

Abstract

Objective The rising incidence of oral tongue squamous cell carcinoma (OTSCC) in patients who have never smoked and the paucity of knowledge of its biological behavior prompted us to develop a new cell line originating from a never-smoker. Materials and methods Fresh tumor tissue of keratinizing OTSCC was collected from a 44-year-old woman who had never smoked. Serum-free media with a low calcium concentration were used in cell culture, and a multifaceted approach was taken to verify and characterize the cell line, designated UCSF-OT-1109. Results UCSF-OT-1109 was authenticated by STR DNA fingerprint analysis, presence of an epithelial marker EpCAM, absence of human papilloma virus (HPV) DNA, and SCC-specific microscopic appearance. Sphere-forming assays supported its tumorigenic potential. Spectral karyotype (SKY) analysis revealed numerical and structural chromosomal abnormalities. Whole-exome sequencing (WES) identified 46 non-synonymous and 13 synonymous somatic single-nucleotide polymorphisms (SNPs) and one frameshift deletion in the coding regions. Specifically, mutations of CDKN2A, TP53, SPTBN5, NOTCH2, and FAM136A were found in the databases. Copy number aberration (CNA) analysis revealed that the cell line loses chromosome 3p and 9p, but lacks amplification of 3q and 11q (as does HPV-negative, smoking-unrelated OTSCC). It also exhibits four distinctive focal amplifications in chromosome 19p, containing 131 genes without SNPs. Particularly, 52 genes showed >3- to 4-fold amplification and could be potential oncogenic drivers. Conclusion We have successfully established a novel OTSCC cell line from a never-smoking patient. UCSF-OT-1109 is potentially a robust experimental model of OTSCC in never-smokers.

Original languageEnglish (US)
Pages (from-to)1-10
Number of pages10
JournalOral Oncology
Volume69
DOIs
StatePublished - Jun 1 2017
Externally publishedYes

Keywords

  • Cell line
  • DNA copy number aberration
  • Insertion or deletion
  • Never-smoker
  • Oral tongue squamous cell carcinoma
  • Single nucleotide polymorphism
  • SKY analysis
  • Sphere-forming assay
  • STR DNA fingerprint analysis
  • Whole-exome sequencing

ASJC Scopus subject areas

  • Oral Surgery
  • Oncology
  • Cancer Research

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