葫芦素B通过激活线粒体途径诱导人肺癌NCI-H460细胞凋亡

Cucurbitacin B induces apoptosis of NCI-H460 lung cancer cells through activation of mitochondrial pathway

目的探讨葫芦素B对人肺癌NCI-H460细胞增殖及凋亡的影响,并探讨其机制。方法体外培养NCI-H460细胞,MTT法观察葫芦素B对细胞增殖的影响,倒置相差显微镜观察细胞形态的变化,采用流式细胞术检测细胞凋亡率。采用罗丹明123(Rhodamine 123)染色,通过流式细胞仪检测线粒体膜电位(△ψm),采用Western blot方法检测线粒体内和胞浆内的细胞色素C。结果葫芦素B对NCI-H460细胞的增殖有抑制作用,且呈剂量、时间依赖性;葫芦素B处理组细胞形态发生显著改变,细胞皱缩、变圆,可见胞核染色质浓缩;流式细胞仪检测结果显示葫芦素B诱导NCI-H460细胞凋亡,呈剂量依赖性。葫芦素B处理后线粒体膜电位显著下降,细胞色素C由线粒体释放到胞浆中。结论葫芦素B可以抑制人肺癌NCI-H460细胞的增殖并诱导细胞凋亡,线粒体途径参与葫芦素B诱导的NCI-H460细胞凋亡。

Objective To study the influence of Cucurbitacin B on the proliferation and apoptosis of NCIH460 lung cancer cells and investigate the mechanism. Methods The NCI-H460 cells were cultured in vitro. The growth inhibitory effect of Cucurbitacin B on NCI-H460 cells was detected by M TT assay. The morphological changes were observed under inverted microscope. Apoptosis rate was determined by flowcytometry analysis. The mitochondrial membrane potential（ Δψm） was detected by flowcytometry following rhodamine 123 staining. The expression of mitochondrial cytochrome C and cystolic cytochrome C was determined by Western blot. Results The growth of NCIH460 cells was inhibited by Cucurbitacin B in a dose-and time-dependent manner. M arked morphological changes such as cell volume shrinking and chromatin condensation were observed after treatment by Cucurbitacin B. Flowcytometry analysis showed that Cucurbitacin B could induce NCI-H460 cells apoptosis in a dose-dependent manner. Cucurbitacin B treatment induced significant disruption of △ψm. Cytochrome C was released from mitochondria to cytoplasm. Conclusion Cucurbitacin B could inhibit the growth of NCI-H460 lung cancer cells and induce cell apoptosis through activation of mitochondrial pathway.