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Oral Oncol. 2017 Jun;69:1-10. doi: 10.1016/j.oraloncology.2017.03.020. Epub 2017 Apr 3.

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

Author information

1
Department of Otolaryngology-Head and Neck Surgery, School of Medicine, University of California, San Francisco, San Francisco, CA 94143, USA; UCSF Helen Diller Family Comprehensive Cancer Center, School of Medicine, University of California, San Francisco, San Francisco, CA 94143, USA.
2
UCSF Helen Diller Family Comprehensive Cancer Center, School of Medicine, University of California, San Francisco, San Francisco, CA 94143, USA.
3
Department of Pathology, School of Medicine, University of California, San Francisco, San Francisco, CA 94143, USA.
4
Department of Pathology and Laboratory Medicine, Roswell Park Cancer Institute, Buffalo, NY 14263, USA.
5
Department of Laboratory Medicine, School of Medicine, University of California, San Francisco 94143, USA.
6
UCSF Helen Diller Family Comprehensive Cancer Center, School of Medicine, University of California, San Francisco, San Francisco, CA 94143, USA; Department of Epidemiology & Biostatistics, School of Medicine, University of California, San Francisco 94143, USA.
7
Department of Otolaryngology-Head and Neck Surgery, School of Medicine, University of California, San Francisco, San Francisco, CA 94143, USA; UCSF Helen Diller Family Comprehensive Cancer Center, School of Medicine, University of California, San Francisco, San Francisco, CA 94143, USA. Electronic address: Osamu.Tetsu@ucsf.edu.

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.

KEYWORDS:

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

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