Dimethylfumarate induces cell cycle arrest and apoptosis via regulating intracellular redox systems in HeLa cells
Dimethylfumarate induces cell cycle arrest and apoptosis via regulating intracellular redox systems in HeLa cells
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DOI：
10.1007/s11626-016-0069-2
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发表时间：
2016-12-01
影响因子：
2.1
通讯作者：
Zhou, Qiang
中科院分区：
文献类型：
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作者：
Han, Guocan;Zhou, Qiang
富马酸二甲酯 (DMF) 对多种细胞具有细胞毒性，可作为抗肿瘤药物。本研究旨在探讨DMF对宫颈癌细胞的影响及其潜在机制。培养HeLa细胞并分别用0、50、100、150和200μM DMF处理。 24小时后，使用细胞计数试剂盒-8（CCK-8）测定评估细胞生长，并使用流式细胞术检查细胞周期。此外，Annexin V/碘化丙啶（PI）染色检测细胞凋亡，Western blotting检测caspase-3和聚ADP核糖聚合酶（PARP）的表达。然后评估氧化还原相关因素。此外，在DMF和N-乙酰-L-半胱氨酸（NAC，氧自由基清除剂）联合处理后，HeLa细胞中检测到了所有指标。 DMF 明显抑制 HeLa 细胞数量和细胞生长，且呈剂量依赖性，细胞周期停滞在 G0/G1 期（P < 0.05）。 HeLa细胞凋亡明显增加，caspase-3和PARP表达水平显着增加，且呈DMF浓度依赖性（P < 0.05）。同时，DMF处理后发现a-I-3m损失，活性氧和O-2（中心点-）增加，过氧化氢酶活性和谷胱甘肽（GSH）水平降低（P < 0.05）。添加NAC后，所有这些变化均显着减弱，甚至完全消失（P < 0.05）。总之，DMF对HeLa细胞增殖和凋亡的细胞毒性主要与细胞内GSH耗竭的细胞内氧化还原系统有关。
Dimethylfumarate (DMF) is cytotoxic to several kinds of cells and serves as an anti-tumor drug. This study was designed to investigate the effects and underlying mechanism of DMF on cervical cancer cells. HeLa cells were cultured and treated with 0, 50, 100, 150, and 200 mu M DMF, respectively. After 24 h, cell growth was evaluated using Cell Counting Kit-8 (CCK-8) assay and the cell cycle was examined using flow cytometry. In addition, cell apoptosis was detected by Annexin V/propidium iodide (PI) staining and the expressions of caspase-3 and poly-ADP-ribose polymerase (PARP) were detected using western blotting. The redox-related factors were then assessed. Furthermore, all of the indicators were detected in HeLa cells after combined treatment of DMF and N-acetyl-l-cysteine (NAC, an oxygen-free radical scavenger). The cell number and cell growth of HeLa were obviously inhibited by DMF in a dose-dependent manner, as the cell cycle was arrested at G0/G1 phase (P < 0.05). The apoptotic HeLa cells were markedly increased, and the expression levels of caspase-3 and PARP were significantly increased in a DMF concentration-dependent way (P < 0.05). Meanwhile, loss of a-I-3m, increase in reactive oxygen species and O-2 (center dot-), and the decrease in catalase activity and glutathione (GSH) level were found after DMF treatment (P < 0.05). All these changes were significantly attenuated and even completely disappeared by adding NAC (P < 0.05). In conclusion, the cytotoxicity of DMF on cell proliferation and apoptosis of HeLa cells was mainly related to the intracellular redox systems by depletion of intracellular GSH.