三氧化二砷对黑素瘤B_(16)细胞内活性氧自由基的影响

Influence of As_2O_3 on the reactive oxygen species in melanoma B_(16) cells

目的：探讨三氧化二砷（As2O3）对黑素瘤B16细胞增殖及细胞内活性氧自由基（ROS）水平的影响。方法：30μmol/LAs2O3与500μmol/L N-乙酰半胱氨酸（N-acetylcysteine,NAC）分别单独或联合作用于B16细胞,应用四甲基偶氮唑蓝（MTT）法测定细胞生长抑制情况;流式细胞术检测ROS水平。结果：As2O3能明显抑制B16细胞的增殖,NAC可部分拮抗As2O3对黑素瘤细胞的杀伤效应及诱导凋亡效应。As2O3作用1 h后的B16细胞内ROS水平比对照组升高（P〈0.01）,4 h达到高峰,24 h有所下降,与对照组差异均有统计学意义（P〈0.01）。ROS升高的趋势可被抗氧化物NAC部分阻断（P〈0.05-P〈0.01）。结论：As2O3诱导B16细胞凋亡与ROS水平改变有关,As2O3可通过提高细胞内ROS水平诱导细胞凋亡而发挥抗肿瘤作用,这也可能是As2O3抗黑素瘤的途径之一。

Objective：To study the influence of As2O3 on the proliferation and the reactive oxygen species （ROS） levels in melanoma B16 ceils. Methods ：B16 ceils were treated with 30 μmol/L As2O3 alone or together with 500 μmol/L N-acetylcysteine （NAC）. The effect of As2O3 on B 16 cells was determined by MTY assay, and the intracellular ROS level was measured by flow cytometry （FCM）. Results： As203 could obviously inhibit the proliferation of B16 cells and NAC might partially antagonise the apoptosis effect induced by As2O3. ROS level in B16 cells started to rise after being incubated with 30μmol/L As203 for 1 h（P 〈0.01 ） ,and reached the peak point of at 4 h（ P 〈 0.01 ）, and the ROS expression levels decreased slightly at 24 h. The difference was all significant compared with the control group（ P 〈 0.01 ）. The increasing of ROS might be partially blocked by NAC （ P 〈 0.05 to P 〈 0.01 ）. Conclusions ： B16 cells apoptosis induced by As203 is related to the increase of ROS level. As2O3 induces B16 ceils apoptosis by accumulation of intracellular ROS level, which may be one of the antitumor processes of As2O3.