A Novobiocin Derivative, XN4, Inhibits the Proliferation of Chronic Myeloid Leukemia Cells by Inducing Oxidative DNA Damage.
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.
Department of Pharmacology, School of Pharmacy, Fujian Medical University (FMU), Fuzhou, P. R.China; Institute of Materia Medica, FMU, Fuzhou, P. R.China; Fuijan Key Laboratory of Natural Medicine pharmacology, FMU, Fuzhou, P. R.China.
Lixian
L
Wu
Fujian Institute of Hematology, Union Hospital, FMU, Fuzhou, P. R.China.
Xianling
X
Chen
Department of Pharmacology, School of Pharmacy, Fujian Medical University (FMU), Fuzhou, P. R.China; Institute of Materia Medica, FMU, Fuzhou, P. R.China; Fuijan Key Laboratory of Natural Medicine pharmacology, FMU, Fuzhou, P. R.China.
Lisen
L
Huang
Department of Pharmacology, School of Pharmacy, Fujian Medical University (FMU), Fuzhou, P. R.China; Institute of Materia Medica, FMU, Fuzhou, P. R.China; Fuijan Key Laboratory of Natural Medicine pharmacology, FMU, Fuzhou, P. R.China.
Jue
J
Tian
Department of Pharmacochemistry, School of Pharmacy, FMU, Fuzhou, P. R.China.
Fang
F
Ke
Department of Pharmacology, School of Pharmacy, Fujian Medical University (FMU), Fuzhou, P. R.China; Institute of Materia Medica, FMU, Fuzhou, P. R.China; Fuijan Key Laboratory of Natural Medicine pharmacology, FMU, Fuzhou, P. R.China.
Jianhua
J
Xu
Fujian Institute of Hematology, Union Hospital, FMU, Fuzhou, P. R.China.
Yuanzhong
Y
Chen
Department of Anatomy, School of Basic Medicine, FMU, Fuzhou, P. R.China.
Ming
M
Zheng
D018952:Antigens, CD34 / Q000235:genetics / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D002453:Cell Cycle / Q000187:drug effects / Q000235:genetics; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects / Q000235:genetics; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics; D004249:DNA Damage / Q000187:drug effects* / Q000235:genetics; D005434:Flow Cytometry; D006801:Humans; D020014:K562 Cells; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D000074663:NADPH Oxidase 4; D000074664:NADPH Oxidase 5; D019255:NADPH Oxidases / Q000235:genetics / Q000378:metabolism; D009675:Novobiocin / Q000031:analogs & derivatives / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction
D018952:Antigens, CD34; D008565:Membrane Proteins; D017382:Reactive Oxygen Species; D009675:Novobiocin; D000074663:NADPH Oxidase 4; D000074664:NADPH Oxidase 5; D019255:NADPH Oxidases; C498910:NOX4 protein, human; C431571:NOX5 protein, human