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Characterization of a Novel Tumorigenic Esophageal Adenocarcinoma Cell Line: OANC1

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Abstract

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.

Aim

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 β-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.

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Acknowledgments

The authors would like to thank the Peter MacCallum Cancer Centre Department of Pathology for performing immunohistochemistry, FISH and sequencing, the Histology Core in the Research Division for preparation of xenograft tissue blocks, and Dr. James Eshleman for the provision of JH-EsoAd1 cells. The patient history and fresh and FFPE tumour tissue used in this project was provided by the Victorian Cancer Biobank with appropriate ethics approval. The Victorian Cancer Biobank is supported by the Victorian Government, Australia. NJC and WAP are supported by grants from the National Health and Medical Research Council, Australia. This work was supported by grants from the National Health and Medical Research Council, Australia.

Conflict of interest

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Authors and Affiliations

  1. Surgical Oncology Research Laboratory, Peter MacCallum Cancer Centre, Locked Bag#1, A’Beckett Street, Melbourne, VIC, 8006, Australia

    Nicholas J. Clemons, Christina Fennell & Wayne A. Phillips

  2. Department of Pathology, Peter MacCallum Cancer Centre, Locked Bag#1, A’Beckett Street, Melbourne, VIC, 8006, Australia

    Hongdo Do, Siddhartha Deb, Andrew Fellowes & Alexander Dobrovic

  3. Sir Peter MacCallum Department of Oncology, University of Melbourne, Melbourne, VIC, Australia

    Nicholas J. Clemons, Hongdo Do, Siddhartha Deb, Alexander Dobrovic & Wayne A. Phillips

  4. Department of Pathology, University of Melbourne, Melbourne, VIC, Australia

    Alexander Dobrovic

  5. Department of Surgery, St. Vincent’s Hospital, University of Melbourne, Melbourne, VIC, Australia

    Nicholas J. Clemons & Wayne A. Phillips

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  1. Nicholas J. Clemons
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Correspondence to Nicholas J. Clemons.

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Clemons, N.J., Do, H., Fennell, C. et al. Characterization of a Novel Tumorigenic Esophageal Adenocarcinoma Cell Line: OANC1. Dig Dis Sci 59, 78–88 (2014). https://doi.org/10.1007/s10620-013-2882-8

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  • DOI: https://doi.org/10.1007/s10620-013-2882-8

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