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Characterisation and validation of a new oesophageal adenocarcinoma cell line (#305)

Nicholas J Clemons 1 2 3 , Christina M Fennell 1 , Siddhartha Deb 1 , Wayne A Phillips 1 2 3
  1. Peter MacCallum Cancer Centre, East Melbourne, VIC, Australia
  2. Sir Peter MacCallum Department of Oncology, University of Melbourne, Melbourne, VIC, Australia
  3. Department of Surgery - St. Vincent's Hospital, University of Melbourne, Melbourne, VIC, Australia

Basic and translational research into oesophageal adenocarcinoma (OAC) has been hampered by a relative lack of representative cell lines, particularly when compared with other solid tumours such as ovarian cancer and melanoma. This was compounded in 2010 by the revelation that three commonly used OAC cell lines were derived from other tumour types (1).
We have isolated a new OAC cell line (OACNC1) from a moderately differentiated polypoid OAC resected from a 44 year old Caucasian male. The patient had completed neo-adjuvant chemotherapy and radiotherapy two months prior to surgery. Pathological analysis of the resection specimen showed that the tumour had extended into the muscularis propria, but not through the entire wall, and had spread to the local lymph nodes. There was focal intestinal metaplasia (Barrett’s oesophagus) with areas of low and high grade dysplasia proximal to the tumour.
Tumour cells were isolated from a fresh piece of the OAC following digestion in dispase and trypsin. Microsatellite based genotyping of the cell line DNA and patient DNA confirmed the origin of the cell line. The cell line was tumourigenic in SCID mice, with tumours reaching >1000mm3 two to three months after sub-cutaneous injection. In comparison, OE33 cells injected in SCID mice formed tumours that were approximately 40mm3 after three months. OACNC1 xenograft tumours were highly proliferative (>10 mitoses/high-powered field), poorly differentiated adenocarcinomas that were positive for keratins 7, 8/18 and 19, SOX9 and CDX2 but negative for CK20, MUC2 and MUC5AC.
Thus, we have established, verified and characterised a new OAC cell line that exhibits more vigorous growth in SCID mice than the widely used OE33 cell line. This new cell line will be a useful tool for researchers studying both basic and translational aspects of this disease.

1. Boonstra JJ, van Marion R, Beer DG, Lin L, Chaves P, Ribeiro C, Pereira AD, Roque L, Darnton SJ, Altorki NK, Schrump DS, Klimstra DS, Tang LH, Eshleman JR, Alvarez H, Shimada Y, van Dekken H, Tilanus HW, and Dinjens WN. Verification and unmasking of widely used human esophageal adenocarcinoma cell lines. J Natl Cancer Inst 102: 271-274, 2010.