ROLE OF MANGANESE SUPEROXIDE DISMUTASE IN CHEMOTHERAPY- INDUCED OXIDATIVE STRESS By

......................................................................................... 10 CHAPTER ONE: INTRODUCTION................................................................................ 11 AGGRESSIVE NON-HODGKIN LYMPHOMAS.................................................. 11 Diffuse large B-cell lymphoma............................................................ 12 Mantle cell lymphoma..................................................................... 13 REDOX ENVIRONMENT......................................................................... 14 Redox environment in cancer............................................................. 16 Chemotherapy and oxidative stress...................................................... 17 Role of MnSOD as a tumor suppressor.................................................. 17 REGULATION OF MnSOD BY C/EBPβ AND FOXO TRANSCRIPTION FACTORS........ 20 MAMMALIAN TARGET OF RAPAMYCIN PATHWAY ......................................... 21 USE OF mTOR INHIBITORS IN TREATMENT OF LYMPHOMA.............................. 23 STATEMENT OF THE PROBLEM................................................................ 25 CHAPTER TWO: METHODS.................................................................... 27 REAGENTS AND DRUG TREATMENT......................................................... 27 CELL CULTURE & TRANSFECTIONS.......................................................... 27 Cell Lines................................................................................. 27 C/EBPβ LIP Construction................................................................ 27 Transfections.............................................................................. 28 PROTEIN MEASUREMENTS..................................................................... 28 CELL VIABILITY AND APOPTOSIS MEASUREMENTS....................................... 28 Eosin Y Staining......................................................................... 28 Caspase-3 Activity....................................................................... 29 MTS Assay................................................................................ 29 ROS MEASUREMENTS......................................................................... 29 DCF Measurements..................................................................... 29 Amplex Red Measurements............................................................ 39 MitoSOX Measurements................................................................ 39 GLUTATHIONE MEASUREMENTS............................................................... 31 IMMUNOBLOTS.................................................................................. 31 NUCLEAR RUN-ON ASSAY AND REAL-TIME PCR.......................................... 31 STATISTICS....................................................................................... 32 SNP ANALYSIS.................................................................................. 32