Characterization of a novel tumorigenic esophageal adenocarcinoma cell line: OANC1.
BACKGROUND
Esophageal adenocarcinoma (EAC) has a very high case fatality rate and is one of the fastest rising cancers worldwide. At the same time, research into EAC has been hampered by a relative lack of pre-clinical models, including representative cell lines.


OBJECTIVE
The purpose of this study was to establish and characterize a new EAC cell line.


METHODS
Tumor cells were isolated from EAC tissue by enzymatic digestion. Origin of the cell line was confirmed by microsatellite based genotyping. A panel of cancer-related genes was screened for mutations by targeted deep sequencing, Sanger sequencing and high resolution melting.CDKN2A promoter methylation was assessed by methylation specific high resolution melting. HER2 amplification was assessed by fluorescent in situ hybridization. Immunohistochemistry was used to assess expression of markers in xenografts grown in SCID mice.


RESULTS
A novel EAC cell line, OANC1, was derived from a Barrett's-associated EAC. Microsatellite-based genotyping of OANC1 and patient DNA confirmed the origin of the cell line. Sequencing of OANC1 DNA identified homozygous TP53 missense (c.856G[A, p.E286K)and SMAD4 nonsense (c.1333C[T, p.R445X) mutations.OANC1 are tumorigenic when injected sub-cutaneously into SCID mice and xenografts were positive for columnar, glandular and intestinal epithelial markers commonly expressed in EAC. Xenografts exhibited strong p53 expression, consistent with a TP53 mutation. Some proteins, including p16, EGFR and b-catenin, had heterogeneous expression patterns across xenograft cross-sections, indicative of tumor heterogeneity.


CONCLUSIONS
OANC1 represents a valuable addition to the limited range of pre-clinical models for EAC. This new cell line will be a useful model system for researchers studying both basic and translational aspects of this disease.
Nicholas J
NJ
Clemons
Hongdo
H
Do
Christina
C
Fennell
Siddhartha
S
Deb
Andrew
A
Fellowes
Alexander
A
Dobrovic
Wayne A
WA
Phillips
D000230:Adenocarcinoma / Q000473:pathology*; D000818:Animals; D063646:Carcinogenesis; D045744:Cell Line, Tumor / Q000737:chemistry / Q000473:pathology* / Q000637:transplantation; D004252:DNA Mutational Analysis; D004938:Esophageal Neoplasms / Q000473:pathology*; D016158:Genes, p53; D060005:Genotyping Techniques; D006801:Humans; D051379:Mice; D016513:Mice, SCID; D051901:Smad4 Protein / Q000235:genetics; D023041:Xenograft Model Antitumor Assays
C497796:SMAD4 protein, human; D051901:Smad4 Protein