Using RNA interference in lung cancer cells to target the IKK-NF-κB pathway.
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.
Department of Biochemistry, Chemistry Institute, University of São Paulo, nº1280 - Butantã, São Paulo, SP, 05508-070, Brazil, basseres@iq.usp.br.
Daniela S
DS
Bassères
Albert S
AS
Baldwin
D045744:Cell Line, Tumor; D049109:Cell Proliferation; D015870:Gene Expression; D015972:Gene Expression Regulation, Neoplastic; D055785:Gene Knockdown Techniques; D017930:Genes, Reporter; D006801:Humans; D051550:I-kappa B Kinase / Q000235:genetics / Q000378:metabolism*; D008175:Lung Neoplasms / Q000235:genetics* / Q000378:metabolism*; D016328:NF-kappa B / Q000378:metabolism*; D034622:RNA Interference; D015398:Signal Transduction; D014162:Transfection
D016328:NF-kappa B; D051550:I-kappa B Kinase