目的：研究青蒿素（Artemisinin）对非小细胞肺癌（non-small cell lung cancer cell，NSCLC）细胞（ASTC-a-1和A549细胞）凋亡的影响，并初步探讨其作用机制。方法：CCK-8法检测0~150 μg/ml的青蒿素处理24 h和100 μg/ml青蒿素处理0、6、12、24、48 h对ASTC-a-1和A549细胞活性的影响；激光共聚焦显微镜观察1 μmol/L STS、100 μg/ml青蒿素单独或联合5 mmol/L 活性氧簇（reactive oxygen species,ROS)清除剂NAC（N-乙酰半胱氨酸）处理细胞24 h对细胞核形态的影响，并用流式细胞术检测青蒿素单独或联合NAC对细胞凋亡的影响；通过RNA干扰技术沉默Bax和Bak基因后，观察青蒿素对细胞活性的影响。结果：青蒿素能够以浓度和时间依赖的方式抑制ASTC-a-1和A549细胞的活性，IC50值约为100 μg/ml；经NAC预处理后，青蒿素导致的细胞活性下降程度明显低于未经预处理的细胞，差异有统计学意义（均P<0.01）。STS、青蒿素单独或联合NAC均能引起ASTC-a-1和A549细胞核固缩及细胞凋亡。经NAC预处理后，青蒿素诱导的ASTC-a-1和A549细胞凋亡率分别为（20.4±2.1）%和（17.9±3.8）%，明显低于STS组\[（48.2±2.6）%，（39.8±4.9）%\]和细胞未经预处理组\[（59.6±3.4）%，（50.7±3.8）%\]青蒿素引起的凋亡率（均P<0.01）。青蒿素只能引起Bak沉默细胞活性的上升（P<0.01），而对Bax沉默细胞活性没有明显影响（P>0.05）。结论：青蒿素能诱导两种非小细胞肺癌细胞（ASTC-a-1和A549细胞）ROS介导的细胞凋亡，促凋亡因子Bak而不是Bax参与了凋亡过程。

Objective:To investigate the effect of Artemisinin on apoptosis of non-small cell lung cancer (NSCLC) ASTC-a-1 and A549 cells and its mechanism. Methods:CCK-8 assay was performed to assess the effect of treatment with Artemisinin at 0-150 μg/ml for 24 h and at 100 μg/ml for 0, 6, 12, 24, 48 h on the activity of ASTC-a-1 and A549 cells. Laser scanning confocal microscope was used to observed influence of treatment with STS at 1 μg/ml, Artemisinin at 100 μg/ml alone or combined with NAC for 24 h on nuclear morphologies, and flow cytometry was used to examine impact of treatment with Artemisinin alone or combined with NAC on apoptosis of the cells; After silencing of Bax and Bak genes by RNA interference technology, effect of Artemisinin on viability of the cells was examined.Results: Artemisinin induced growth inhibition of ASTC-a-1 and A549 cells in a concentration and time dependent manner (IC50≈100 μg/ml), and the falling level of viability of the cells induced by Artemisinin was significantly less than untreaded cells after pretreated with NAC (all P<0.01). Treatments of ASTC-a-1 and A549 cells with STS, Artemisinin alone or combined with NAC induced their karyopyknosis and apoptosis. After pretreatment with NAC, the apoptosis rates of ASTC-a-1 and A549 cells induced by Artemisinin were (20.4±2.1)% and (17.9±3.8)% respectively, which were obviously less than STS group ( \[48.2±2.6\]% and \[39.8±4.9\]%, respectively) and without pretreated group (\[59.6±3.4\]% and \[50.7±3.8\]% respectively) (all P<0.01). Artemisinin only increased viability of silencing Bak cells (P<0.01), but not obviously impact that of silencing Bax cells (P＞0.05). Conclusion: Artemisinin induces apoptosis of non-small cell lung cancer ASTC-a-1 and A549 cells, which requires the participation of reactive oxygen species (ROS) and promoting apoptosis factor Bak, but not Bax.

河南省医学科技攻关计划项目（No. 2011020091）