青藤碱通过线粒体途径诱导肺癌NCI-H460细胞凋亡的初步研究

The primary research on sinomenine induced apoptosis of lung cancer NCI- H460 cells by the mitochondrial signaling pathway

目的:研究青藤碱(sinomenine,SIN)对人肺癌NCI-H460细胞株的生长抑制及诱导凋亡作用及其机制。方法:四甲基偶氮噻蓝(MTT)法检测细胞增殖的抑制作用,流式细胞仪Annexin V/PI双染法检测细胞凋亡,Td T酶介导的d UTP缺口末端标记(TUNNEL)方法观察细胞的凋亡,罗丹明123(Rhodamine123)染色流式细胞仪检测线粒体膜电位(ΔΨm)。结果:SIN对NCI-H460细胞株生长具有抑制作用并诱导凋亡。Annexin V/PI双染检测细胞的凋亡可见随SIN浓度增加,细胞凋亡增加呈浓度依赖性。TUNNEL阳性的凋亡细胞呈棕黄色,细胞核片段化改变。罗丹明染色检测线粒体膜电位的结果提示在SIN作用于NCI-H460细胞48h后,线粒体膜电位下降,SIN浓度越高,膜电位下降越显著。结论:SIN具有明显的细胞毒作用,能诱导NCIH460细胞凋亡,SIN通过线粒体途径诱导NCI-H460细胞凋亡。

Objective: To study the effects of sinomenine on the growth and apoptosis of human lung cancer cell NCI- H460 and its mechanisms involved in the process. Methods: The effect of sinomenine on the proliferation of NCI- H460 cell was determined by MTT assay. The apoptosis of NCI- H460 cells was analyzed by Annexin V-FITC and PI double staining. Morphological changes of cells were observed by TUNNEL assay. The mitochondrial membrane potential( ΔΨm) was examined by flow cytometry with Rhodamine123 staining. Results: Sinomenine could inhibit proliferation and induce apoptosis of NCL- H460 cell. The proportion of apoptosis cells in sinomenine- treated cells were increased in a dose- dependent manner which was demonstrated in the Annexin V- FITC and Pl double staining assay. The apoptosis cells with TUNEL positive were stained with brown- yellow color and the DNA fragmentation was observed in these cells. The result of Rhodamine123 staining indicated that sinomenine treatment could induce significant disruption of Δψm in a dose- dependent manner in case of sinomenine treatment for 48 h. Conclusion: Sinomenine has obvious cytotoxic effect and could induce the apoptosis of NCI- H460 cells. Sinomenine induced NCI- H460 cells apoptosis by mitochondrial pathway.