细梗香草总皂苷诱导肺癌细胞H460凋亡及其机制的研究

Capillipes inhibits proliferation and induces apoptosis of human lung cancer H460 cells

目的观察细梗香草总皂苷(LC)对非小细胞肺癌H460细胞增殖和凋亡的影响。方法在H460肺癌细胞培养时加入不同浓度的LC,采用MTT法和克隆形成实验检测细胞的增殖活性,AnnexinV/PI(FCM)法观察LC对肺癌细胞凋亡的诱导作用和周期分布的影响。DCFH-DA荧光探针FCM检测细胞内活性氧生成,Western bIot法检测NF-K B、I—kB、Bax、Bcl-2、Capase-3的蛋白表达。结果 LC可抑制H460细胞生长,具有剂量、时间依赖性;LC作用H460细胞后克隆形成率明显下降。LC可引起H460细胞凋亡,并可将H460生长阻滞在S期。LC以浓度依赖方式增高H460细胞内活性氧水平,还可使H460细胞l—k B、Bax和Capase-3表达增加,而使NF-k B和BcI-2表达降低。结论LC可以抑制H460细胞增殖,可能是通过增加细胞内活性氧表达,激活了肺癌细胞线粒体凋亡途径。

Objective To investigate the effect of capil iposides isolated from Lysimachia capil ipes Hemsl（capil ipes, LC） on proliferation and apoptosis of human non- smal lung cancer cellline H460 in vitro. Methods H460 cells were cultured with different concentration of LC. The proliferation, colony formation and cellcycle of H460 cells were measured by MTT method, colony formation assay and flow cytometry, respectively. Intracellular reactive oxygen species （ROS） was measured by FCM with fluorescent probe of DCFH- DA. The expression of signal transduction proteins NF- κB, I- κB, Bax, Bcl- 2 and capase- 3 was detected by Western blotting. Results The proliferation of H460 cells was inhibited and the ability of colony- formation was re-duced;the apoptosis rate of H460 cells was increased（P〈0.05）and the cellcycle was arrested at S phase（P〈0.05）after cul-tured with LC for 24h. LC significantly increased ROS level in H460 cells in a concentration- dependent manner. LC up- regulated I- κB and Bax expression,and activated capase- 3 enzyme （P〈0.05）while the expression levels of NF- ΚB and Bcl- 2 were down- regulated（P〈0.05）. Conclusion LC can inhibit the growth and induce apoptosis of H460 cells, which may be associated with ROS generation and activation of mitochondrial apoptotic pathway.