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Dermatology - Molecular Biology - Oncology

Irreversible EGFR Inhibitor EKB-569 Targets Low-LET ?-Radiation-Triggered Rel Orchestration and Potentiates Cell Death in Squamous Cell Carcinoma
Published: Thursday, December 29, 2011
Author: Natarajan Aravindan et al.

by Natarajan Aravindan, Charles R. Thomas, Sheeja Aravindan, Aswathi S. Mohan, Jamunarani Veeraraghavan, Mohan Natarajan

EKB-569 (Pelitinib), an irreversible EGFR tyrosine kinase inhibitor has shown potential therapeutic efficiency in solid tumors. However, cell-killing potential in combination with radiotherapy and its underlying molecular orchestration remain to be explored. The objective of this study was to determine the effect of EKB-569 on ionizing radiation (IR)-associated NF?B-dependent cell death. SCC-4 and SCC-9 cells exposed to IR (2Gy) with and without EKB-569 treatment were analyzed for transactivation of 88 NF?B pathway molecules, NF?B DNA-binding activity, translation of the NF?B downstream mediators, Birc1, 2 and 5, cell viability, metabolic activity and apoptosis. Selective targeting of IR-induced NF?B by EKB-569 and its influence on cell-fate were assessed by overexpressing (p50/p65) and silencing (?I?Ba) NF?B. QPCR profiling after IR exposure revealed a significant induction of 74 NF?B signal transduction molecules. Of those, 72 were suppressed with EKB-569. EMSA revealed a dose dependent inhibition of NF?B by EKB-569. More importantly, EKB-569 inhibited IR-induced NF?B in a dose-dependent manner, and this inhibition was sustained up to at least 72 h. Immunoblotting revealed a significant suppression of IR-induced Birc1, 2 and 5 by EKB-569. We observed a dose-dependent inhibition of cell viability, metabolic activity and apoptosis with EKB-569. EKB-569 significantly enhanced IR-induced cell death and apoptosis. Blocking NF?B improved IR-induced cell death. Conversely, NF?B overexpression negates EKB-569 -induced cell-killing. Together, these pre-clinical data suggest that EKB-569 is a radiosensitizer of squamous cell carcinoma and may mechanistically involve selective targeting of IR-induced NF?B-dependent survival signaling. Further pre-clinical in-vivo studies are warranted.