A Novobiocin Derivative, XN4, Inhibits the Proliferation of Chronic Myeloid Leukemia Cells by Inducing Oxidative DNA Damage
A Novobiocin Derivative, XN4, Inhibits the Proliferation of Chronic Myeloid Leukemia Cells by Inducing Oxidative DNA Damage
复制标题
新生霉素衍生物 XN4 通过诱导氧化 DNA 损伤抑制慢性粒细胞白血病细胞的增殖
DOI：
10.1371/journal.pone.0123314
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发表时间：
2015-04-30
期刊：
影响因子：
3.7
通讯作者：
Zheng, Ming
中科院分区：
文献类型：
--
作者：
Wu, Lixian;Chen, Xianling;Zheng, Ming
XN 4可能通过活性氧（reactive oxygen species，ROS）诱导DNA损伤和细胞凋亡。MTT法检测K562和K562/G 01细胞的增殖抑制作用。通过qRT-PCR评价NADPH氧化酶1-5（Nox 1 -5）基因的mRNA水平。流式细胞仪检测细胞外活性氧（ROS）水平、DNA损伤、细胞凋亡和细胞周期进程。通过免疫印迹分析蛋白质水平。XN 4显著抑制K562和K562/G 01细胞的增殖，IC 50值分别为3.75±0.07 µM和2.63±0.43 µM。XN 4显著增加Nox 4和Nox 5 mRNA的表达，刺激细胞内ROS的产生，诱导DNA损伤，激活ATM-γ-H2 AX信号通路，使细胞周期中处于S期和G2/M期的细胞数量增加。随后，XN 4通过激活caspase-3和PARP诱导凋亡细胞死亡。上述作用可被活性氧清除剂N-乙酰半胱氨酸（NAC）逆转。此外，XN 4可以诱导从CML患者骨髓中分离的祖细胞/干细胞的凋亡。总之，XN 4诱导的CML细胞DNA损伤和细胞凋亡是由ROS的产生介导的。
XN4 might induce DNA damage and apoptotic cell death through reactive oxygen species (ROS). The inhibition of proliferation of K562 and K562/G01 cells was measured by MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide). The mRNA levels of NADPH oxidase 1-5 (Nox1-5) genes were evaluated by qRT-PCR. The levels of extracellular reactive oxygen species (ROS), DNA damage, apoptosis, and cell cycle progression were examined by flow cytometry (FCM). Protein levels were analyzed by immunoblotting. XN4 significantly inhibited the proliferation of K562 and K562/G01 cells, with IC50 values of 3.75±0.07 µM and 2.63±0.43 µM, respectively. XN4 significantly increased the levels of Nox4 and Nox5 mRNA, stimulating the generation of intracellular ROS, inducing DNA damage and activating ATM-γ-H2AX signaling, which increased the number of cells in the S and G2/M phase of the cell cycle. Subsequently, XN4 induced apoptotic cell death by activating caspase-3 and PARP. Moreover, the above effects were all reversed by the ROS scavenger N-acetylcysteine (NAC). Additionally, XN4 can induce apoptosis in progenitor/stem cells isolated from CML patients’ bone marrow. In conclusion, XN4-induced DNA damage and cell apoptosis in CML cells is mediated by the generation of ROS.