洋地黄毒苷诱导小细胞肺癌NCI-H446细胞增殖和凋亡的影响

Effect of Digitoxin on Proliferation and Apoptosis of Human Small Cell Lung Cancer NCI-H446 Cell

应用四甲基偶氮唑盐(MTT)法检测洋地黄毒苷对NCI-H446细胞增殖的抑制效果;用AO-EB染色、DNA凝胶电泳分析以及AnnexinⅤ检测法检测细胞凋亡;用碘化丙啶(PI)染色测定其周期变化,利用激光共聚焦显微镜观察细胞内活性氧(ROS)和钙离子(Ca2+)的变化.结果显示洋地黄毒苷明显抑制NCI-H446细胞增殖,AO-EB染色、DNA电泳及AnnexinV检测法显示细胞有明显的凋亡现象,PI染色显示处于S期的细胞增多,激光共聚焦显微镜观察表明细胞内活性氧和钙离子浓度均升高.表明洋地黄毒苷能明显抑制NCI-H446细胞增殖,且细胞被阻滞在S期.细胞内活性氧和钙离子浓度的增加可能与其诱导细胞凋亡有关.

Growth inhibition was detected by MTT assay. Cell apoptosis was assayed by staining of cell with AO-EB, DNA fragmentation and Annexin V -TITC/PI FCM . Cell cycle was detected by staining of cell with PI, the changes of intracellular reactive oxygen species and free calcium were determined with laser confocal microscopy. The result showed that Digitoxin remarkably inhibited NCI-H446 cells proliferation. As shown by AO-EB fluores- cence staining, DNA fragmentation assay and Annexin V-TITC/PI FCM detection, NCI- H446 ceils apoptosis could be induced by Digitoxin. PI staining assay indicated that the ratio of S phase of NCI-H446 cells was increased. In addition,Digitoxin increased the level of intra- cellular reactive oxygen species and C obviously. That means Digitoxin can inhibit the prolif- eration of NCI-H446 and block the cell cycle progression in S phase. The induced-apoptosis effect may be due to the boost of concentrations of reactive oxygen species and free calcium.