Predictive markers for treatment sensitivity in head and neck squamous cell carcinoma

.............................................................................................................. 7 LIST OF PAPERS ........................................................................................................ 9 ABBREVIATIONS ..................................................................................................... 10 INTRODUCTION ..................................................................................................... 11 Cancer ............................................................................................................ 11 Cancer of the head and neck ........................................................................... 11 Treatment ....................................................................................................... 13 Therapeutic decision making ..................................................................... 13 Surgery ..................................................................................................... 13 Radiation .................................................................................................. 13 Chemotherapy .......................................................................................... 14 Chemoirradiation ...................................................................................... 14 EGFR-targeted therapy ............................................................................... 15 Cellular processes influencing treatment sensitivity .......................................... 16 Apoptosis ................................................................................................. 16 Cell cycle regulation .................................................................................. 17 DNA-repair ................................................................................................ 18 Single Nucleotide Polymorphisms (SNPs) ................................................... 19 Predictive markers .......................................................................................... 20 Survivin ..................................................................................................... 20 Members of the Bcl-2 family ...................................................................... 21 Heat shock protein 70 (Hsp70) .................................................................. 22 The epidermal growth factor receptor (EGFR) ............................................ 23 Cyclin D1 .................................................................................................. 25 Cyclooxygenase-2 (COX-2) ........................................................................ 26 SMAD4 ...................................................................................................... 26 p53 ........................................................................................................... 27 Murine double minute 2 (MDM2) ............................................................... 28 Fibroblast growth factor receptor 4 (FGFR4) .............................................. 30 SNPs in the DNA repair genes XPC, XPD, XRCC1, and XRCC3 ..................... 31 Other predictive markers ........................................................................... 32 AIMS OF THE THESIS .............................................................................................. 35 MATERIAL AND METHODS...................................................................................... 37 Ethical aspects .......................................................................................... 37 Cells and culture conditions ...................................................................... 37 The Linköping HNSCC biobank .................................................................. 37 Assessment of intrinsic radiosensitivity ..................................................... 38 Assessment of intrinsic cisplatin sensitivity ............................................... 39 Western blot .............................................................................................. 40 ELISA ......................................................................................................... 40 Restriction fragment length polymorphism (RFLP) ..................................... 41 Pyrosequencing of MDM2 .......................................................................... 41 Single stranded conformation analysis (SSCA) ........................................... 42 Number of Negative Points (NNP) .............................................................. 43 Statistical methods .................................................................................... 43