Abstract Pancreatic adenocarcinoma (PDAC) is the fourth leading cause of cancer-related death in the UK. Five-year survival rates are less than 3% and haven’t improved over the last 40 years. Thus novel approaches are required urgently. We have found that high expression of the integrin β6 subunit gene (ITGB6) correlates with a significant reduction (Hazard Ratio 1.74; Logrank p=6x10-4) in overall survival from PDAC. Thus we have begun to examine the biological functions mediated by αvβ6 in a transgenic model of PDAC. A panel of cell lines were generated from PDAC tumors of a novel transgenic mouse and characterized for markers of tissue identity, integrin expression and ability to invade using 3D-organotypic assays. Cell populations appeared to contain both epithelial (E-cadherin positive) and mesenchymal (vimentin positive) characteristics, highlighting their polyclonality. In addition, two cell populations, KVC1 and KPCD3, expressed low levels of αvβ6 and were studied further. KVC1 and KPCD3 cells were labeled with 10D5 (anti-αvβ6) and sorted by FACS to obtain high-αvβ6 expressing monoclonal cell lines. The resulting cell lines after four rounds of sorting (S4) demonstrated mean MFI values of approximately 4.5-fold (KVC1 vs. KVC1 S4 A+) and approximately 3-fold (KPCD3 vs. KPCD3 S4 E+) higher than their parental counterparts. To compare the invasion profile of the parental populations and S4 monoclonal cell lines, cells were placed in 3D organotypic assays with mouse embryonic fibroblasts (MEFs) for 14 days before gels were fixed and processed to paraffin. H&E stained sections revealed that parental populations KVC1 and KPCD3 formed invasive, single-cell walled ductal structures that had penetrated into the matrigel-collagen mix, but only in the presence of MEFs. In contrast, αvβ6-positive S4 populations, KVC1 S4 A+ and KPCD3 S4 E+, also demonstrated strong invasion but the invasive ductal phenotype was lost, the cells invading as small cohesive groups and individual cells. These data suggest that the pattern of PDAC tumour invasion may be modulated by expression of the integrin αvβ6. Current studies explore the mechanisms by which these phenotypic changes are generated. Citation Format: Claire S. Reader, Sabari Vallath, Jennifer Morton, Colin Steele, Hemant M. Kocher, Stephen M. Keyse, Owen Sansom, John F. Marshall. Cell lines generated from PDAC transgenic mice reveal integrin αvβ6 modifies the invasive phenotype of pancreatic cancer. [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Innovations in Research and Treatment; May 18-21, 2014; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2015;75(13 Suppl):Abstract nr B87.