Aim To investigate the effect of Novobiocin(Nov) on its cytotoxicity against CML cells, and to explore its underlying mechanisms by which Nov might induce DNA damage and apoptosis through reactive oxygen species (ROS). Methods MTT and CFSE were used to measure the proliferation inhibition ratio of K562 and K562/G01 cells; Flow cytometry (FCM) was used to teste the level of extracellular ROS, DNA damage, cell cycle progression, mitochondria membrane potential (MPP) and apoptosis; Western blot was used to verify the amount of proteins. Result Nov significantly inhibited the proliferation of CML cells with increased the generation of intracellular ROS, followed by induction of DNA damage, activation of the ATM-p53-r-H2AX pathway and checkpoint-related signals CHK1/CHK2, which led to increased numbers of cells in the S and G2/M phases of the cell cycle. Furthermore, Nov induced apoptosis through decrease of mitochondrial membrane potential and activation of caspase-3 and PARP. The above effects were all prevented by the ROS scavenger N-acetylcysteine. Conclusion These findings therefore suggest that Nov induced DNA damage and mitochondria-dependent cell apoptosis in CML cells are mediated via ROS generation.
国家科技期刊平台
NSTL
维普
万方数据
