Differential Activity of the KRAS Oncogene by Method of Activation: Implications for Signaling and Therapeutic Intervention

摘要: v Table of contents vii List illustrations xii tables xvi abbreviations xvii Chapter 1: Introduction 1 1.1 Ras background and function 1.1.1 The activation cycle 2 1.1.2 localization 5 1.1.3 KRAS in development 7 1.2 KRas signal transduction 8 1.3 cancer 11 1.3.1 Activating mutations viii 1.3.2 as a driver tumorigenesis 12 1.3.3 therapy 13 2: Materials methods 15 2.1 BATTLE clinical trial 2.2 Patient microarray data 2.3 Cells culture conditions 17 2.4 Preparation protein lysates reverse-phase array (RPPA) 2.5 Immunoblots immunoprecipitations 18 2.6 Viral vector construction 19 2.7 Soft agar growth assays 2.8 Molecular modeling 2.9 Screening compounds against isogenic mutant lines 20 2.10 NSCLC cell line panel 2.11 siRNA screening 2.12 Individual plasmid transfection 2.13 Spheroid formation 21 2.14 Anoikis assay 22 2.15 Cell analysis