摘要
目的:探讨皖南五步蛇蛇毒抑瘤组分Ⅰ(AAVC-1)诱导人非小细胞型肺癌A549细胞凋亡的作用,并分析其可能的线粒体机制。方法:以不同作用浓度的AAVC-1处理A549细胞,采用Annexin V/PI双染流式细胞仪分析法检测A549细胞凋亡百分率;JC-1检测细胞线粒体的膜电位变化,免疫组化检测细胞色素C在线粒体内的分布。结果:不同浓度梯度AAVC-1 1.0、3.0、9.0、20.0μg/m L处理24 h后,与对照组相比,A549细胞早期凋亡率明显增加(P<0.05);线粒体膜电位绿色荧光百分比由(23.01±6.07)%上升至(87.26±9.54)%(P<0.01),荧光显微镜观察到绿色荧光增强;免疫组化显示,随着AAVC-1浓度增加,胞质内细胞色素C平均光密度值明显增加(P<0.05)。结论:AAVC-1可以通过降低线粒体膜电位和促进细胞色素C的释放,激活线粒体通路诱导人非小细胞型肺癌A549的凋亡。
Objective: To investigate the apoptosis pathway and mechanisms of human non-small cell lung cancer A549 cells induced by anticoagulant fraction from Agkistrodon acutus venom( AAVC-1).Methods: Different dose of AAVC-1 was used to treat A549 cells,and the apoptosis percentage of A549 cells was assessed by flow cytometry with annexin V / PI double staining. Mitochondrial trans-membrane potential( Δψm) of A549 cells was measured by JC-1 fluorescence probe and microscopy,and distribution of cytochrome C in the mitochondria was detected by immunohistochemistry after AAVC-1 treatment.Results: The early apoptosis rate of A549 cells was significantly increased after treatment with different dose of AAVC-1 for 24 h as compared with the control group( P〈0.05).The fluorescence green intensity was increased from( 20.13±1.6) % to( 89.12± 1.2) %( P〈0.01),and fluorescence spectroscopy indicated intensified intracellular green fluorescence.Immunohistochemical test revealed markedly increased average light density of cytochrome C with added dose of AAVC-1 treatment( P〈0.05).Conclusion: AAVC-1 can induce the apoptosis of human non-small cell lung cancer A549 cells by reducing the mitochondrial membrane potential and promoting the release of cytochrome C.
作者
徐成
张根葆
周淑艳
桑金凤
XU Cheng ZHANG Genbao ZHOU Shuyan SANG Jinfeng(School of Clinical Medicine, Wanan Medical College, Wuhu 241002, China)
出处
《皖南医学院学报》
CAS
2017年第1期5-7,15,共4页
Journal of Wannan Medical College
基金
皖南医学院大学生科研资助项目(RG20151204)
安徽高校省级自然科学研究重点项目(KJ2011A266)
关键词
五步蛇毒抑瘤组分Ⅰ
非小细胞型肺癌
线粒体
细胞凋亡
Agkistrodon acutus venom antitumor component 1
non-small cell lung cancer
mitochondria
apoptosis