Abstract Growth factor and K-ras-induced generation of reactive oxygen species have important roles in mediating pancreatic ductal adenocarcinoma. Pancreatic cancer cells also express high levels of active transcription factor Nuclear Factor-κB (NF-κB), which is responsible for cell survival and resistance to chemotherapeutics. The serine/threonine kinase Protein Kinase D1 (PKD1) is a sensor for oxidative stress and protects cancer cells by inducing anti-apoptotic and anti-oxidant genes. Here we provide evidence that PKD1 is the means of how high NF-κB activity levels are achieved in pancreatic cancer cells. We also show the contribution of K-ras, which is upregulated in its activity in over 90% of PDAC (pancreatic ductal adenocarcinoma). We found that K-ras in pancreatic ductal HPDE cells increases PKD1 activity and that the depletion of the pancreatic ductal adenocarcinoma cell line Panc-1 from K-ras leads to decreased PKD1 activity and NF-κB activation. As a mechanism we found that K-ras mediates activation of PKD1 via Src and that these signaling events relay to high NF-κB levels. Our data reveal K-ras as an activator of PKD1 and show that both oncogenes, Src and K-ras cooperate to activate NF-κB in PDAC through PKD1. This suggests the K-ras/Src/PKD1/NF-κB pathway as a new potential drug target for pancreatic cancer. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3143.