Using RNA interference in lung cancer cells to target the IKK-NF-κB pathway.

Abstract

RNA interference-based gene silencing has become a widely used technology to evaluate how inhibition of expression of individual proteins affects biological readout. Through the use of this technology, a lot has been learned about how different proteins function in a wide variety of biological contexts, including cancer. In this context, RNA interference-mediated gene silencing has contributed to further our understanding of how different proteins in the NF-κB signaling pathway (including the NF-κB members themselves) contribute to cancer. Here, we describe two RNA interference-based protocols in lung cancer cells targeting upstream activators of NF-κB transcription factor: the catalytic subunits of the IKK complex. The first protocol is designed to evaluate the impact of IKKα or IKKβ inhibition on NF-κB transcriptional activity, whereas the second protocol is designed to evaluate how siRNA-mediated IKK inhibition affects lung cancer cell proliferation.