Analysis of epidermal growth factor-induced NF-κB signaling.
The nuclear factor kappaB (NF-κB) is a family of transcription factors that control cell survival, cell proliferation, cell differentiation, inflammatory responses, and innate and adaptive immune responses. Its activation is tightly regulated, and incorrect regulation of NF-κB has been linked to a variety of pathological diseases, including cancer initiation and progression. NF-κB is often constitutively activated in cancer cells to promote cell survival, proliferation, migration, and/or epithelial-to-mesenchymal transition (EMT). Although the mechanism of constitutive NF-κB activation in cancer cells is not fully understood, it has been shown that mutation or aberrant expression of epidermal growth factor receptor (EGFR) contributes to this, and the NF-κB activation, in turn, contributes to cell proliferation, survival, metastasis, and drug resistance in various cancers. Recent study from our lab indicates that CARMA3, similar to the function of CARMA1 in mediating antigen receptor-mediated NF-κB activation, plays an essential role in mediating EGFR-induced NF-κB activation. However, the mechanism on how EGFR induces NF-κB activation is not clearly understood. In this chapter, we describe the methods required to test and characterize the role of a potential signaling component in EGFR-induced NF-κB activation.
Division of Basic Science Research, Department of Molecular and Cellular Oncology, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Blvd, Unit 108, Houston, TX, 77030, USA.
Changying
C
Jiang
Xin
X
Lin
D015153:Blotting, Western / Q000379:methods; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D024202:Electrophoretic Mobility Shift Assay / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D058750:Epithelial-Mesenchymal Transition; D066246:ErbB Receptors / Q000378:metabolism; D055785:Gene Knockdown Techniques; D006801:Humans; D047468:Immunoprecipitation / Q000379:methods; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism*; D011485:Protein Binding; D034741:RNA, Small Interfering / Q000235:genetics; D015398:Signal Transduction / Q000187:drug effects*; D014162:Transfection
D016328:NF-kappa B; D034741:RNA, Small Interfering; D004815:Epidermal Growth Factor; D008156:Luciferases; D066246:ErbB Receptors