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目的研究青蒿琥酯(artesunate,ART)对人子宫颈癌细胞HeLa凋亡与增殖的影响。方法通过MTT法观察ART对HeLa细胞的生长抑制作用;荧光染色观察ART作用前后细胞形态的改变;免疫组化检测HeLa细胞caspase-3表达;流式细胞术观察ART对细胞周期的影响,并检测细胞线粒体膜电位ΔΨm和细胞内活性氧(reactive oxygen species,ROS)的改变。结果ART对HeLa细胞的生长抑制作用呈时间、剂量依赖关系;荧光染色观察见凋亡小体形成;透射电镜见细胞染色质凝聚,边集呈新月形;免疫组化检测细胞质caspase-3呈阳性表达;流式细胞术检测ART可引起HeLa细胞周期S期阻滞;细胞内ROS明显升高(P<0.01),ΔΨm显著降低(P<0.05)。结论ART作用于人子宫颈癌HeLa细胞,可能通过线粒体途径诱导细胞凋亡,抑制细胞增殖。
Objective To study the effect of artesunate (ART) on the apoptosis and proliferation of human cervical carcinoma HeLa cells. Methods MTT assay was used to observe the inhibitory effect of ART on HeLa cells. Fluorescence staining was used to observe the changes of cell morphology before and after ART treatment. The expression of caspase-3 in HeLa cells was detected by immunohistochemistry. The cell cycle was observed by flow cytometry Mitochondrial membrane potential ΔΨm and intracellular reactive oxygen species (ROS) changes. Results ART inhibited the growth of HeLa cells in a time-and dose-dependent manner. The formation of apoptotic bodies was observed by fluorescence staining. The chromatin was aggregated and the crescent was observed by transmission electron microscopy. The expression of caspase-3 in cytoplasm was detected by immunohistochemistry (P <0.01), and ΔΨm was significantly decreased (P <0.05). The results of flow cytometry showed that ART could induce the arrest of S phase in HeLa cell cycle. Conclusion ART acts on human cervical cancer HeLa cells, which may induce cell apoptosis through mitochondrial pathway and inhibit cell proliferation.