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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 proliferation inhibitory effect of pachouli alcohol (PA) on human androgen-independent prostate cancer cell line DU145 and to explore its mechanism. Methods: patchouli alcohol (10,20,40,80,160 mg · L-1) was applied to detect the proliferation of DU145 cells at a density of 6 × 107 / L for 24, 48 and 72 h. The drug-induced morphological changes of apoptosis were observed by transmission electron microscopy. The changes of drug-induced apoptosis were detected by flow cytometry with Annexin V-FITC and PI double labeling method. The levels of cysteine Caspase-3, Bcl-2, Bcl-2 and Livin. Results: MTT assay showed that patchouli alcohol treatment inhibited the cell proliferation significantly. The inhibitory rates of 10, 20, 40, 80 and 160 mg · L -1 patchouli alcohol on DU145 were 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% in a time-and dose- ). Transmission electron microscopy showed apoptotic morphological changes of cells after drug treatment. Flow cytometry showed that patchouli alcohol effect significantly increased the rate of apoptosis, compared with the control group was statistically significant (P <0.05). Western blot showed that patchouliol enhanced the expression of Caspase-3, Bax and down-regulated the expressions of Livin and Bcl-2. Conclusions: Patchouli alcohol can inhibit the proliferation of DU145 cells, which may be related to the induction of apoptosis.