香鳞毛蕨苷提取物E对人肺癌A549细胞增殖及凋亡的影响

Effect of Compound E from Dryopteris Fragrans(L.) Schott on the Proliferation and Apoptosis of Human Lung Cancer A549 Cells

目的:研究香鳞毛蕨苷提取物E(Fragranoside E)对人肺癌A549细胞增殖及凋亡的影响。方法:将体外培养的人肺癌A549细胞分为对照组(0.1%DMSO、100 nM紫杉醇)和实验组,通过CCK-8实验、克隆形成实验检测细胞增殖情况;检测乳酸脱氢酶(LDH)水平以探讨Fragranoside E的细胞毒性;透射电镜及流式细胞术检测A549细胞的凋亡情况。进一步采用5mM N-乙酰半胱氨酸(N-acetylcysteine,NAC)预处理A549细胞2 h后,采用荧光显微镜观察细胞内ROS的释放情况。结果:CCK8及克隆形成实验结果提示Fragranoside E呈浓度和时间依赖性抑制A549细胞增殖(P<0.05);≤40μM Fragranoside E处理A549细胞对其LDH的释放无显著影响(P>0.05);而40μM Fragranoside E可诱导A549细胞凋亡,细胞内ROS升高,NAC(5 m M)预处理2 h后,细胞内ROS(112.6%±12.3%)较Fragranoside E处理组显著降低(P<0.05)。结论:Fragranoside E能够抑制A549细胞增殖,诱导其凋亡,其抗肿瘤活性可能与细胞内ROS释放有关。

Objective: To investigate the efftect of Fragranoside E from Dryopteris fragrans(L.) Schott on the proliferation and apoptosis of human lung cancer of A549 cells. Methods: Human lung cancer A549 cells cultured in vitro were divided into the control group(0.1 %DMSO, 100 n M paclitaxel) and the Fragranoside E group. Cell proliferation was examined by Cell counting kit(CCK-8) assay and clone formation assay;cytotoxicity was tested by LDH assay;cell apoptosis was detected by transmission electron microscope and flow cytometry assay;releasing of ROS was observed by fluorescence microscopy, after some cells were pretreated with N-acetylcysteine(NAC) for 2 h. Results: The results of CCK8 and clone formation experiments suggested that Fragranoside E inhibited the proliferation of A549 cells in a dose-dependent and time-dependent manner(P<0.05), without any obvious cytotoxic below 40 μM concentration in LDH assay. Fragranoside E(40 μM) induced A549 cell apoptosis and the increasing of ROS, which was significantly reduced(112.6 %±12.3 %) in cells pretreated with NAC(P<0.05). Conclusion: Fragranoside E inhibited the proliferation and induced the apoptosis of A549 cells, which might be closely related to the releasing of ROS.