摘要
目的:观察广藿香醇(pachouli alcohol,PA)对人雄激素非依赖性前列腺癌细胞DU145的凋亡诱导效应及增殖抑制作用,探讨其发生机制。方法:广藿香醇10,20,40,80,160 mg·L-1作用在密度为6×107/L DU145细胞24,48,72 h后,采用四甲基偶氮唑盐法(MTT)检测广藿香醇对细胞的生长抑制作用;透射电镜观察药物诱导细胞凋亡形态变化;Annexin V-FITC,PI双标记法流式细胞仪检测药物诱发细胞凋亡的改变;Western blot检测半胱氨酸天冬氨酸蛋白酶3(Caspase-3)、B细胞淋巴瘤/白血病-2蛋白(Bcl-2)、Bcl-2关联X蛋白(Bax)、凋亡抑制蛋白Livin的表达。结果:MTT法检测提示广藿香醇处理组细胞增殖抑制作用明显,10,20,40,80,160 mg·L-1广藿香醇对DU145的24,48,72 h抑制率分别为5.73%,16.67%,22.61%;13.42%,25.03%,39.68%;25.92%,31.46%,52.76%;39.61%,54.68%,73.52%;56.42%,77.35%,91.58%,呈剂量-时间依赖性(P<0.05)。透射电镜显示药物作用后细胞出现凋亡形态改变。流式细胞仪检测提示广藿香醇作用使细胞凋亡率明显升高,与对照组比较有统计学意义(P<0.05)。Western blot检测提示广藿香醇可增强细胞Caspase-3,Bax的表达,下调Livin,Bcl-2蛋白的表达。结论:广藿香醇能抑制DU145细胞增殖,其作用机制可能与诱导细胞凋亡效应有关。
Objective： To observe the apoptosis inducing effect and inhibitory effect of pachouli alcohol on the proliferation of human androgen independent prostate cancer cell line DU145, and its mechanisms. Method： After prostate cancer DU145 cells were treated with pachouli alcohol for 24, 48, 72 h, the apoptosis morphology was observed by transmission electron microscope;cell viability was estimated using MTT assay;cell apoptosis rate was measured by flow cytometry. Western blot was used to test Caspase-3, Bcl-2, Bax and Livin protein expression. Result： Apoptosis of DU145 cells was detected by transmission electron microscopy after using pachouli alcohol. In vitro, pachouli alcohol significantly inhibited the proliferation of DU145 cells in dose and time-dependent manners which was detected by MTT method. The apoptosis rate was increased significantly which compare with the control group. Pachouli alcohol could enhance Caspase-3, Bax protein expression, and reduce Livin, Bcl-2 protein expression in DU145 cells. Conclusion： Pachouli alcohol can inhibit the proliferation of DU145 in vitro, which may be correlated with inducing the cell apoptosis.
出处
《中国实验方剂学杂志》
CAS
北大核心
2014年第10期165-169,共5页
Chinese Journal of Experimental Traditional Medical Formulae
基金
四川省教育厅科学研究项目(13ZB0324)
关键词
广藿香醇
人前列腺癌细胞DU145
增殖
细胞色素C途径
凋亡
pachouli alcohol
human prostate cancer cell line DU145
proliferation
cytochrome C pathway
apoptosis
