放射生物学的核心.ppt

Ionizing Radiation-Induced Checkpoint and DNA RepairYa Wang, Ph.D.(12-2004)Thomas Jefferson UniversityPhiladelphia, USA Cell Cycle Checkpoints: Definition Checkpoints are biochemical pathways. Checkpoints regulate the progression, in terms of order and timing, of cells from one phase of the cell cycle to the next (normal checkpoint). Checkpoints slow down the progression of cells through the cycle after DNA damage (DNA damage-induced checkpoint) ATM and ATR are two of the most important checkpoint proteins in mammalian cells. Importance of Checkpoint Two major DNA DSBs repair pathways exist in mammalian cells Does checkpoint activation facilitate both NHEJ and HRR? A1-5 cells show an extreme radioresistance than B4 cells A1-5 and B4 cells have the similar rejoining abilities of DSBs A1-5 cells show a stronger G2 checkpoint response than B4 cells. A1-5 cells show the stronger response of S checkpoint than B4 cells. A1-5 cells have a higher expression of CHK1 than B4 cells Chk1 but not Chk2 antisense oligonucleotide abolishes the stronger S checkpoint of A1-5 cells Chk1 but not Chk2 antisense oligonucleotide abolishes the stronger G2 checkpoint and the radioresistance in A1-5 cells Two major DNA DSBs repair pathways exist in mammalian cells Caffeine had no effect on NHEJ of CHO cells synchronized in G1 phase Caffeine Radiosensitized the NHEJ Proficient and Deficient Cells HR: Eukaryotic Genes Caffeine abolished IR-induced S checkpoint response Caffeine had a great radiosensitizing effect on wild type as well as ATM-/-cells but had less effect on Rad54-/- cells and had no effect on Rad51 deficient cells. A model for caffeine radiosensitization Acknowledgments Xinag Wang Xiang-Yang Zhou Checkpoint inhibition does not directly affect NHEJ but radiosensitizes cells. Does checkpoint activation facilitate HRR, therefore, radiosensitize cells? Rad51, Rad52, Rad54, the Rad51 paralogs: XRCC2, XRCC3, Rad51B, Rad51C, Rad51D and Rad50 complex (Rad50,