桔梗皂苷-D诱导肺腺癌A549和A549/DDP细胞凋亡和周期阻滞

被引:7
|
作者
赵若琳
周坤福
张旭
陈美娟
机构
[1] 南京中医药大学江苏省中医药防治肿瘤协同创新中心
关键词
桔梗皂苷-D(PD); 非小细胞肺癌; 凋亡; 周期阻滞; A549; A549/DDP;
D O I
10.13192/j.issn.1000-1719.2015.08.044
中图分类号
R285 [中药药理学];
学科分类号
摘要
目的:探究桔梗皂苷-D(Platycodin-D,PD)诱导肺腺癌细胞(A549)和A549顺铂耐药细胞(A549/DDP)凋亡和周期抑制的作用机制,为PD治疗非小细胞肺癌和产生顺铂耐药的非小细胞肺癌提供实验基础。方法:采用MTT法检测细胞的增殖抑制率和耐药指数;流式细胞术检测细胞周期阻滞和凋亡作用;Hoechst-33258荧光染色观察细胞核形态的变化;Western-blot蛋白印迹法分析凋亡通路和细胞周期相关蛋白表达水平的变化。结果:MTT检测结果显示,PD对A549和A549/DDP细胞的增殖抑制呈浓度和时间依赖性。Hoechst 33258荧光染色发现,随着PD浓度增加,两种细胞核形态都有明显的改变。进一步流式细胞术结果表明,PD诱导A549和A549/DDP细胞凋亡和周期阻滞。Westernblot检测结果表明,PD通过上调Bax、Caspase-9、cleaved-Caspase-3和PARP的表达,下调Bcl-2和Caspase-3的表达诱导A549和A549/DDP细胞凋亡。此外,PD通过下调Cyclin-D1、Cyclin-D3、CDK4、CDK6、E2F1和p-Rb及上调P18的蛋白水平诱导A549细胞周期G0/G1阻滞。同时,PD通过下调Cyclin-B1、p-Cdc2、E2F1和p-Rb及上调P27的蛋白表达诱导A549/DDP细胞周期G2/M阻滞。结论:PD通过线粒体途径诱导A549和A549/DDP细胞凋亡诱导,并且引发A549细胞G0/G1周期阻滞和A549/DDP细胞G2/M阻滞周期阻滞。
引用
收藏
页码:1484 / 1489+1603 +1603
页数:7
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