Mahmoud Toulany1, Klaus Dittmann, Michael Baumann, H Peter Rodemann. 1. Division of Radiobiology and Molecular Environmental Research, Department of Radiation Oncology, Eberhard-Karls University Tuebingen, Roentgenweg 11, 72076 Tuebingen, Germany.
Abstract
BACKGROUND AND PURPOSE: To investigate the cellular and molecular consequences of antagonizing radiation-induced EGFR-activation in vitro. PATIENTS AND METHODS: The effect of the EGFR tyrosine kinase inhibitor BIBX1382BS on radiation sensitivity was determined after single- and fractionated-dose irradiation in human cell lines of bronchial carcinoma (A549), breast adeno-carcinoma (MDA-MB-231), pharyngeal squamous-cell carcinoma (FaDu), squamous-cell carcinoma of cervix (HTB-35) as well as normal (HSF-7) and transformed (HH4-DED) human skin fibroblasts. Applying immuno-precipitation and western blotting pattern of radiation-dependent activation of different components of EGFR-signaling after pre-treatment with and without BIBX1382BS or other tyrosine kinase inhibitors was analyzed. RESULTS: Autophosphorylation of EGFR which occurred 1-5 min after irradiation (IR, 2 Gy) or treatment with EGF (100 ng/ml) could be inhibited in all cells tested by pre-treatment with BIBX1382BS for 30 min. Combination of drug treatment with fractionated irradiation (4x2 Gy) led to a strong radiosensitizing effect in Ras-mutated A549 and MDA-MB-231 cells, but not in normal Ras presenting cell lines FaDu and HTB-35 or normal and transformed human skin fibroblasts. Both BIBX1382BS as well as the PI3 kinase inhibitor LY294002 led to a blockage (for A549 cells) or reduction (for FaDu cells) of radiation-induced P-AKT. In contrast to FaDu cells, treatment of A549 cells with LY294002 resulted in a significant decrease of post-irradiation survival of A549 cells. Furthermore, only in Ras-mutated cells, but not in normal Ras cells clonogenic survival and phosphorylation of AKT was sensitive to pre-treatment with TGF-alpha-neutralizing antibody indicating an important role of TGF-alpha in regulating radiation-induced EGFR signaling. CONCLUSIONS: Enhancement of radiation sensitivity by the specific EGFR-tyrosine kinase inhibitor BIBX1382BS is not generally achieved in human tumor cells, but depends most likely on the Ras genotype of the cell lines tested.
BACKGROUND AND PURPOSE: To investigate the cellular and molecular consequences of antagonizing radiation-induced EGFR-activation in vitro. PATIENTS AND METHODS: The effect of the EGFR tyrosine kinase inhibitor BIBX1382BS on radiation sensitivity was determined after single- and fractionated-dose irradiation in human cell lines of bronchial carcinoma (A549), breast adeno-carcinoma (MDA-MB-231), pharyngeal squamous-cell carcinoma (FaDu), squamous-cell carcinoma of cervix (HTB-35) as well as normal (HSF-7) and transformed (HH4-DED) human skin fibroblasts. Applying immuno-precipitation and western blotting pattern of radiation-dependent activation of different components of EGFR-signaling after pre-treatment with and without BIBX1382BS or other tyrosine kinase inhibitors was analyzed. RESULTS: Autophosphorylation of EGFR which occurred 1-5 min after irradiation (IR, 2 Gy) or treatment with EGF (100 ng/ml) could be inhibited in all cells tested by pre-treatment with BIBX1382BS for 30 min. Combination of drug treatment with fractionated irradiation (4x2 Gy) led to a strong radiosensitizing effect in Ras-mutated A549 and MDA-MB-231 cells, but not in normal Ras presenting cell lines FaDu and HTB-35 or normal and transformed human skin fibroblasts. Both BIBX1382BS as well as the PI3 kinase inhibitor LY294002 led to a blockage (for A549 cells) or reduction (for FaDu cells) of radiation-induced P-AKT. In contrast to FaDu cells, treatment of A549 cells with LY294002 resulted in a significant decrease of post-irradiation survival of A549 cells. Furthermore, only in Ras-mutated cells, but not in normal Ras cells clonogenic survival and phosphorylation of AKT was sensitive to pre-treatment with TGF-alpha-neutralizing antibody indicating an important role of TGF-alpha in regulating radiation-induced EGFR signaling. CONCLUSIONS: Enhancement of radiation sensitivity by the specific EGFR-tyrosine kinase inhibitor BIBX1382BS is not generally achieved in humantumor cells, but depends most likely on the Ras genotype of the cell lines tested.
Authors: M Saki; M Toulany; W Sihver; M Zenker; J-M Heldt; B Mosch; H-J Pietzsch; M Baumann; J Steinbach; H P Rodemann Journal: Strahlenther Onkol Date: 2012-08-10 Impact factor: 3.621
Authors: Mahmoud Toulany; Minjmaa Minjgee; Mohammad Saki; Marina Holler; Friedegund Meier; Wolfgang Eicheler; H Peter Rodemann Journal: Cancer Biol Ther Date: 2013-12-09 Impact factor: 4.742
Authors: Agata I Rembielak; Pooja Jain; Andrew S Jackson; Melanie M Green; Gillian R Santorelli; Gillian A Whitfield; Adrian Crellin; Angel Garcia-Alonso; Ganesh Radhakrishna; James Cullen; M Ben Taylor; Ric Swindell; Catharine M West; Juan Valle; Azeem Saleem; Patricia M Price Journal: Transl Oncol Date: 2014-02-01 Impact factor: 4.243