¹Cytokine Research Laboratory, Department of Experimental Therapeutics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA

Correspondence: Professor BB Aggarwal, Cytokine Research Laboratory, Department of Experimental Therapeutics, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Blvd, Box 143, Houston, TX 77030, USA. E-mail: aggarwal@mdanderson.org

²Current address: Dr MM Chaturvedi, The Department of Zoology, University of Delhi, Delhi 110007, India.

Received 4 January 2007; Revised 26 March 2007; Accepted 10 April 2007; Published online 28 May 2007.

Abstract

Overexpression of epidermal growth factor (EGF) receptor and constitutive activation of nuclear factor-B (NF-B) are frequently encountered in tumor cells. Although EGF has been shown to induce NF-B activation, the mechanism is poorly understood. EGF activated NF-B DNA binding, induced NF-B reporter activity and the expression of antiapoptotic and cell-proliferative gene products. Interestingly, non-small cell lung adenocarcinoma cell lines (HCC827 and H3255), which exhibit EGFR amplification, showed ligand-independent activation of NF-B. Unlike tumor-necrosis factor (TNF), however, EGF failed to induce IB phosphorylation and ubiquitination and the activation of IB kinase (IKK). Although DN-IKK inhibited TNF-induced NF-B activity, DN-IKK had no effect on EGF-induced NF-B activation, suggesting that EGF-induced NF-B activation is IKK independent. Using dominant-negative plasmids, we also demonstrated the role of TRADD, TRAF2, NIK and Ras in EGF-induced NF-B activation. By using specific antibodies and IB plasmid, which is mutated at tyrosine 42 to phenylalanine, we show that EGF induced the tyrosine phosphorylation of IB at residue 42. Furthermore, EGF receptor kinase inhibitor blocked IB phosphorylation and consequent NF-B activation. Overall, our results indicate that tyrosine phosphorylation of IB at residue 42 is critical for EGF-induced NF-B activation pathway.