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Establishment and Characterization of a Crohn`S Related Colonic Carcinoma Cell Line and Matched Xenograft
Florian Kuehn*, Ernst Klar, Claudia Maletzki, Michael Linnebacher
Dept. of General, Thoracic, Vascular and Transplantation Surgery, University of Rostock, Rostock, Germany

Background & Aims: Colitis-associated colorectal cancer (CAC) seems to be a rather unique entity and differs in some of its genetic alterations, characteristics of tumor formation and clinical features from those of sporadic colorectal carcinoma (CRC). Most of the descriptions about tumor biology of colitis-associated carcinomas are referring to ulcerative colitis whereas data on Crohn`s colitis related carcinoma are scarce. Additionally, the majority of patients with Crohn`s disease are under immunosuppressive treatment; thus generating a different environment for tumor growth and especially for tumor immune escape. Recently, a worse clinical outcome with a decreased 5-year survival rate for patients with Crohn`s related carcinoma compared to sporadic CRC was shown. Representative pre-clinical models of CAC are thus mandatory for identifying molecular characteristics and biomarkers and ultimately for development of novel drugs and therapy regimen.
Methods: Here, we describe the clinical case of a very fast growing CAC in a long-term immunosuppressed patient with Crohn`s disease and the successful establishment and characterization of a CAC cell line (HROC69) along with its corresponding xenograft.
Results: Molecular characterization revealed that both tumor models (cell line and xenografts) were microsatellite-stable. However, a near diploid DNA-status was observed both by flow cytometry and SNP Array analysis. Mutation analysis revealed mutations in TP53, APC and PTEN (interestingly two mutations in Exons 1 and 5) whereas K-ras and B-raf hotspot mutations were not found. The HROC69 cells were confirmed to be (i) human (PCR), (ii) identical to the original tumor (STR-typing) and (iii) of epithelial origin (EpCAM+, FACS). The cells did not express MHC-class I and II molecules. Upon Interferon-γ stimulation, MHC-class II expression was induced, whereas MHC-class I was not. The cell line doubled in 2.46 to 2.83 days. The chemosensitivity profile of the cell line was analyzed in detail.
Conclusions: This newly established and well-characterized, low-passage cell line and its corresponding xenograft of Crohn`s associated carcinoma provide a useful tool for future investigations on the biological characteristics of a rare and distinct tumor entity. Additionally, matched patient-derived immune cells allow for comparative genetic studies as well as detailed analysis of immune escape mechanisms.


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