ERK signaling promotes IKKe expression and oncogenic functions in pancreatic cancer cells in association with TBK1
A variety of cancers utilize RAS-regulated signaling pathways to promote oncogenic phenotypes. A widely studied example is pancreatic cancer, where mutant KRAS signaling leads to activation of MEK/ERK kinases and downstream signaling which promotes oncogenic mechanisms, including cell proliferation. Importantly, ERK inhibitors have shown efficacy in some cancer clinical trials. Previously, others have studied the effects of the related kinases TBK1 and IKKe in pancreatic cancer where they have been shown to promote cell survival. Here, we show that RAS/MAPK signaling promotes expression of IKKe through control of protein stability and not through control of RNA levels. RNAseq analysis indicate that TBK1 and IKKe contribute to the expression of a subset of ERK-regulated genes. Potentially related to the effects on IKKe, proteomic analysis reveals that ERK functions to stabilize a relatively large set of proteins independent of RNA regulation. Knockdown of IKKe and TBK1 individually does not affect growth of MIA PaCa-2 pancreatic cancer cells, but dual knockdown significantly inhibits MIA PaCa-2 growth which is mediated through cell death. Concurrent silencing or inhibition of both TBK1 and IKKe also reduces tumor sphere growth in MIA PaCa-2 cells, correlating with a loss of stemness pathways found with RNAseq. The data suggest the importance of regulation of IKKe by ERK in pancreatic cancer cells and of the combined oncogenic activity of TBK1 and IKKe. Overall design: RNAseq analysis of MIA PaCa-2 pancreatic cancer cells treated for 24H with the ERK1/2 inhibitor SCH772984, the dual TBK1/IKKepsilon inhibitor Compound 1, or DMSO as a control.