Research Papers: Pathology:
Global gene expression in pseudomyxoma peritonei, with parallel development of two immortalized cell lines
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Darren L. Roberts1,2, Sarah T. O’Dwyer3, Peter L. Stern1, Andrew G. Renehan1,2,3
1Immunology Group, Paterson Institute for Cancer Research, The University of Manchester, Manchester, M20 4BX, UK
2Institute of Cancer Sciences, The University of Manchester, Manchester Academic Health Science Centre, The Christie NHS Foundation Trust, Manchester M20 4BX, UK
3Peritoneal Tumour Service, Department of Surgery, The Christie NHS Foundation Trust, Manchester, M20 4BX, UK
Andrew G. Renehan, e-mail: email@example.com
Keywords: pseudomyxoma peritonei, cell line, characterization, exon array
Received: January 17, 2015 Accepted: January 24, 2015 Published: April 13, 2015
Pseudomyxoma peritonei (PMP) is a rare tumor of appendiceal origin. Treatment is major cytoreductive surgery but morbidity is high. PMP is considered chemo-resistant; its molecular biology is understudied; and presently, there is no platform for pre-clinical drug testing. Here, we performed exon array analysis from laser micro-dissected PMP tissue and normal colonic epithelia. The array analysis identified 27 up-regulated and 34 down-regulated genes: candidate up-regulated genes included SLC16A4, DSC3, Aldolase B, EPHX4, and ARHGAP24; candidate down-regulated genes were MS4A12, TMIGD1 and Caspase-5. We confirmed differential expression of the candidate genes and their protein products using in-situ hybridization and immuno-histochemistry. In parallel, we established two primary PMP cell lines, N14A and N15A, and immortalized with an SV40 T-antigen lentiviral vector. We cross-checked for expression of the candidate genes (from the array analyses) using qPCR in the cell lines and demonstrated that the gene profiles were distinct from those of colorectal tumor libraries and commonly used colon cell lines. N14A and N15A were responsiveness to mitomycin and oxaliplatin. This study characterizes global gene expression in PMP, and the parallel development of the first immortalized PMP cell lines; fit for pre-clinical testing and PMP oncogene discovery.
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