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背景与目的:探讨As2O3(arsenic trioxide,ATO)对HL-60细胞凋亡过程中细胞内活性氧(reactive oxygen species,ROS)水平、NF-κB(nuclear factor kappa B),及C-IAP2(cellular inhibitor of apoptosis proteins 2)的影响。材料与方法:以7.5μmol/L As2O3单独应用及与500μmol/L N-乙酰半胱氨酸(N-acetyl-L-cysteine,NAC)联合作用于HL-60细胞12、24 h后,流式细胞术检测HL-60细胞ROS的产生量;Western blot法检测NF-κB P65核蛋白的变化情况;半定量RT-PCR法检测C-IAP2的相对表达量。结果:7.5μmol/L的As2O3作用HL-60细胞12、24 h后细胞内ROS的生成增加,与阴性对照比较,差异具有统计学意义(P<0.05)。NF-κBP65核蛋白的相对含量分别为49.3%±4.4%和23.1%±2.1%,C-IAP2的相对表达分别为72.9%±5.8%和59.3%±4.4%,较对照组均明显降低(P<0.05)。500μmol/L NAC和7.5μmol/L As2O3共同作用HL-60细胞12、24 h后细胞ROS生成量相对减少,与AS2O3单独作用组比较,差异具有统计学意义(P<0.05);NF-κB P65核蛋白的相对含量分别为65.4%±4.9%和37.1%±3.4%,C-IAP2的相对表达量分别为81.1%±5.8%和73.7%±4.9%。较AS2O3单独作用组均明显增加(P<0.05)。结论:As2O3诱导HL-60细胞凋亡过程中,细胞内ROS生成增加,抑制NF-κB活性,同时下调其靶基因C-IAP2等的表达;NAC能阻断As2O3诱导HL-60细胞凋亡过程中ROS的生成,部分阻断了NF-κB活性的抑制。
BACKGROUND & AIM: To investigate the effects of arsenic trioxide (ATO) on the expression of reactive oxygen species (ROS), nuclear factor kappa B (NF-κB) and C-IAP2 inhibitor of apoptosis proteins 2). MATERIALS AND METHODS: HL-60 cells were treated with 7.5μmol / L As2O3 alone or in combination with 500μmol / L N-acetyl-L-cysteine (NAC) for 12 or 24 hours. The amount of ROS production in HL-60 cells was detected by cytometry. The changes of NF-κB P65 nuclear protein were detected by Western blot. The relative expression of C-IAP2 was detected by semi-quantitative RT-PCR. Results: After treated with 7.5μmol / L As2O3 for 12 or 24 hours, the generation of ROS increased in HL-60 cells. Compared with the negative control, the difference was statistically significant (P <0.05). The relative content of NF-κBp65 nuclear protein was 49.3% ± 4.4% and 23.1% ± 2.1% respectively, and the relative expression of C-IAP2 was 72.9% ± 5.8% and 59.3% ± 4.4% <0.05). Compared with AS2O3 treated group, the ROS production of HL-60 cells treated with 500μmol / L NAC and 7.5μmol / L As2O3 for 12,24 h decreased significantly (P <0.05); NF-κB P65 The relative contents of nuclear proteins were 65.4% ± 4.9% and 37.1% ± 3.4%, respectively. The relative expression levels of C-IAP2 were 81.1% ± 5.8% and 73.7% ± 4.9%, respectively. Compared with AS2O3 alone group were significantly increased (P <0.05). CONCLUSION: As2O3 can induce the apoptosis of HL-60 cells during the apoptosis of HL-60 cells, and increase the intracellular ROS production, inhibit the activity of NF-κB, and down-regulate the expression of its target gene C-IAP2. The generation of ROS partially blocked the inhibition of NF-κB activity.