Accumulation of hydrogen peroxide is an early and crucial step for paclitaxel-induced cancer cell death both in vitro and in vivo
Accumulation of hydrogen peroxide is an early and crucial step for paclitaxel-induced cancer cell death both in vitro and in vivo
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DOI：
10.1002/ijc.21685
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发表时间：
2006-07-01
影响因子：
6.4
通讯作者：
Goldwasser, Francois
中科院分区：
文献类型：
--
作者：
Alexandre, Jerome;Batteux, Frederic;Goldwasser, Francois
紫杉醇与微管结合后导致细胞死亡的细胞内事件仍知之甚少。由于活性氧 (ROS) 通过独立的分子靶标参与抗癌药物的细胞毒性，因此我们探讨了 ROS 在紫杉醇细胞毒性中的作用。在体外将 A549 人肺癌细胞暴露于紫杉醇后 15 分钟内，通过分光荧光法检测到细胞内 O-2(o-) 和 H2O2 水平呈浓度依赖性增加。添加 N-乙酰半胱氨酸 (NAC) 或谷胱甘肽（两种 H2O2 清除剂）可导致紫杉醇 IC50 增加 4 倍。将 NAC 共孵育延迟 4 小时，会导致细胞保护作用降低 3 倍。谷胱甘肽合成抑制剂丁硫氨酸亚砜亚胺可显着增加紫杉醇的细胞毒性和 H2O2 积累，但不会改变 O-2(o-) 水平。与二亚苯基碘共同孵育表明紫杉醇诱导的-O-2(o-) 产生部分与细胞质NADPH 氧化酶活性增加有关。与 caspase 3 和 8 抑制剂同时治疗可增加细胞存活率，但不能阻止 H2O2 的早期积累。为了评估 ROS 在紫杉醇抗肿瘤活性中的作用，给小鼠注射 LLC1 肺癌细胞并用紫杉醇腹腔注射治疗。和/或 NAC。 NAC 消除了紫杉醇在小鼠体内的抗肿瘤活性。总之，H2O2 的积累是紫杉醇诱导癌细胞死亡的早期且关键的步骤，然后细胞才会进入细胞凋亡。这些结果表明ROS在体外和体内参与紫杉醇的细胞毒性。 (c) 2006 Wiley-Liss, Inc.
Intracellular events following paclitaxel binding to microtubules that lead to cell death remain poorly understood. Because reactive oxygen species (ROS) are involved in the cytotoxicity of anticancer agents acting through independent molecular targets, we explored the role of ROS in paclitaxel cytotoxicity. Within 15 min after ill vitro exposure of A549 human lung cancer cells to paclitaxel, a concentration-dependent intracellular increase in O-2(o-) and H2O2 levels was detected by spectrofluorometry. Addition of N-acetylcysteine (NAC) or glutathione, two H2O2 scavenger, induced a 4-fold increase in paclitaxel IC50. Delaying NAC co-incubation by 4 hr, resulted in a 3-fold reduction in cell protection. The glutathione synthesis inhibitor, buthionine sulfoximine significantly increased paclitaxel cytotoxicity and H2O2 accumulation, but did not modify O-2(o-) levels. Co-incubation with diphenylene iodonium suggested that paclitaxel induced-O-2(o-) production was in part associated with increased activity of cytoplasmic NADPH oxidase. Concomitant treatment with inhibitors of caspases 3 and 8 increased cell survival but did not prevent the early accumulation of H2O2. To evaluate the role of ROS in paclitaxel antitumoral activity, mice were injected with LLC1 lung cancer cells and treated with paclitaxel i.p. and/or NAC. The antitumoral activity of paclitaxel in mice was abolished by NAC. In conclusion, the accumulation of H2O2 is an early and crucial step for paclitaxel-induced cancer cell death before the commitment of the cells into apoptosis. These results suggest that ROS participate ill vitro and ill vivo to paclitaxel cytotoxicity. (c) 2006 Wiley-Liss, Inc.