Signaling-pathway-in-response-to-DNA-damage-幻灯片.pptVIP

● DNA 损伤—— 真核细胞的DNA分子处在内环境及外环境DNA损伤 物的不断攻击下，这些DNA损伤物或可直接损伤一个 碱基或打断DNA主链，或使DNA链发生交联。 如，正 常细胞代谢中或受到离子辐射时产生的氧化自由基可 使DNA受损伤。 真核生物经长期进化，已建立了完整的DNA损伤修 复机制，以应对各种代谢及外环境中的DNA损伤因素对DNA的损伤。 DNA 损伤信号传导简图 NA Abstract In response to DNA damage, eukaryotic cells activate ATM-Chk2 and/or ATR-Chk1 to arrest the cell cycle and initiate DNA repair. We show that, in the absence of p53, cells depend on a third cell-cycle checkpoint pathway involving p38MAPK/MK2 for cell-cycle arrest and survival after DNA damage. MK2 depletion in p53-deficient cells, but not in p53 wild-type cells, caused abrogation of the Cdc25A-mediated S phase checkpoint after cisplatin exposure and loss of the Cdc25B-mediated G2/M checkpoint following doxorubicin treatment, resulting in mitotic catastrophe and pronounced regression of murine tumors in vivo. We show that the Chk1 inhibitor UCN-01 also potently inhibits MK2, suggesting that its clinical efficacy results from the simultaneous disruption of two critical checkpoint pathways in p53-defective cells. A-D,DNA损伤诱导 MK2及p38活化。 E. MK2 诱导是p38 依赖的。 F.免疫组化，gama-H2AX,DNA 损伤Marker G.Cisplatin，Camp诱导的MK2为ATR依赖 Doxorubsin 诱导的MK2为ATR/ATM依赖 UV诱导ATR/ATM不依赖的MK2激活 DNA损伤毒物激活P38-MK2 通路 基因毒素诱导的DNA损伤时， DNA损伤毒物激活P38-MK2 通路。 ATM-/- ATR-/- Figure?1.?The p38MAPK/MK2 Pathway Is Activated by DNA-Damaging Drugs (A–D) Kinetics of p38MAPK and MK2 activation. U2OS cells were treated with 10 μM cisplatin (A), 10 μM camptothecin (B), 10 μM doxorubicin (C), or?DMSO control (D) for the indicated times. Cell lysates were probed for total and phosphorylated/activated forms of p38MAPK and MK2 (MK-2) by?Western blotting. β-actin served as a loading control. (E) MK2 activation is p38MAPK dependent. U2OS cells were treated with the p38MAPK-specific inhibitor SB203580 (10 μM) or DMSO vehicle for 30?min prior to exposure to chemotherapeutic drugs as in (A)–(D). Total and phosphorylated/activated p38 and MK2 were determined by immunoblotting as above. (F) Activation of M