Mechanism of apoptosis induced by doxorubicin through the generation of hydrogen peroxide
Mechanism of apoptosis induced by doxorubicin through the generation of hydrogen peroxide
复制标题
DOI：
10.1016/j.lfs.2004.05.040
复制
发表时间：
2005-02-11
期刊：
影响因子：
6.1
通讯作者：
Kawanishi, S
中科院分区：
文献类型：
--
作者：
Mizutani, H;Tada-Oikawa, S;Kawanishi, S
阿霉素（DOX）的主要抗癌作用被认为是由于拓扑异构酶II抑制和自由基的产生。我们之前的研究已经证明TAS-103（一种拓扑异构酶抑制剂）通过NAD（P）H氧化酶激活介导的DNA切割和随后的H2 O2生成诱导细胞凋亡[H. Mizutani等人，J.Biol.Chem.277（2002）30684-30689]。因此，为了阐明DOX是否通过相同的机制作为抗癌药物发挥作用，我们研究了DOX在人白血病细胞系HL-60和H2 O2抗性亚克隆HP 100中诱导细胞凋亡的机制。H2 O2诱导HL-60细胞凋亡的DNA梯状条带形成与H2 O2介导的凋亡途径有关，而H2 O2诱导HL-60细胞凋亡的DNA梯状条带形成与H2 O2介导的凋亡途径有关。流式细胞术显示，H2 O2形成之前，DeltaPsim和caspase-3激活的增加。多聚腺苷二磷酸核糖聚合酶（PARP）和NAD（P）H氧化酶抑制剂可抑制阿霉素诱导的HL-60细胞DNA梯状条带的形成。此外，DOX在1h时显著诱导HL-60细胞中8-氧代-7，8-二氢-2 ' -脱氧鸟苷的形成，这是氧化DNA损伤的指标，但在HP 100细胞中不诱导。与其他拓扑异构酶抑制剂（TAS-103、阿柔比星和氨柔比醇）引起DNA裂解和凋亡的能力相比，DOX诱导的细胞凋亡主要由氧化DNA损伤引发。这些结果表明，DOX的关键凋亡触发因素被认为是由DOX诱导的直接H2 O2产生的氧化DNA损伤，尽管DOX诱导的凋亡可能涉及拓扑异构酶II抑制。这种氧化性DNA损伤通过PARP和NAD（P）H氧化酶活化引起间接H2 O2产生，导致DeltaPsim增加和随后的DOX诱导的细胞凋亡中的caspase-3活化。(C)2004年爱思唯尔公司All rights reserved.
The main anticancer action of doxorubicin (DOX) is believed to be due to topoisomerase II inhibition and free radical generation. Our previous study has demonstrated that TAS-103, a topoisomerase inhibitor, induces apoptosis through DNA cleavage and subsequent H2O2 generation mediated by NAD(P)H oxidase activation [H. Mizutani et al. J. Biol. Chem. 277 (2002) 30684-30689]. Therefore, to clarify whether DOX functions as an anticancer drug through the same mechanism or not, we investigated the mechanism of apoptosis induced by DOX in the human leukemia cell line HL-60 and the H2O2-resistant sub-clone, HP100. DOX-induced DNA ladder formation could be detected in HL-60 cells after a 7 h incubation, whereas it could not be detected under the same condition in HP100 cells, suggesting the involvement of H2O2-mediated pathways in apoptosis. Flow cytometry revealed that H2O2 formation preceded the increase in DeltaPsim and caspase-3 activation. Poly(ADP-ribose) polymerase (PARP) and NAD(P)H oxidase inhibitors prevented DOX-induced DNA ladder formation in HL-60 cells. Moreover, DOX significantly induced formation of 8-oxo-7,8-dihydro-2' -deoxyguanosine, an indicator of oxidative DNA damage, in HL-60 cells at 1 h, but not in HP100 cells. DOX-induced apoptosis was mainly initiated by oxidative DNA damage in comparison with the ability of other topoisomerase inhibitors (TAS-103, arnrubicin and amrubicinol) to cause DNA cleavage and apoptosis. These results suggest that the critical apoptotic trigger of DOX is considered to be oxidative DNA damage by the DOX-induced direct H2O2 generation, although DOX-induced apoptosis may involve topoisomerase II inhibition. This oxidative DNA damage causes indirect H2O2 generation through PARP and NAD(P)H oxidase activation, leading to the DeltaPsim increase and subsequent caspase-3 activation in DOX-induced apoptosis. (C) 2004 Elsevier Inc. All rights reserved.