Zeylenone抑制人前列腺癌PC-3细胞增殖及诱导凋亡作用研究  

Inhibitory effect of Zeylenone on proliferation and apoptosis of human PC-3 cells

作　　者：张蕾蕾[1] 李如意[2] 张丽静[1] 陈丹[1,3] 霍小位 李立勇[1] 曹丽[1] 

机构地区：[1]中国医学科学院北京协和医学院药用植物研究所,北京100193 [2]浙江大学生物医学工程及仪器科学学院,浙江杭州310027 [3]贵阳医学院药理研究室,贵州贵阳550004

出　　处：《中国药理学通报》2013年第6期808-813,共6页Chinese Pharmacological Bulletin

基　　金：国家科技部"重大新药创制"科技重大专项-综合性新药研究开发技术大平台-创新药物研究开发技术平台建设(No 2012ZX09301-002-001;2012ZX09501001-004)

摘　　要：目的研究(4R,5S,6S)-4-苯甲酰氧基-6-[(苯甲酰氧基)甲基]-5,6-二羟基-2-环己烯-1-酮Zeylenone(Zey)对人前列腺癌PC-3细胞的生长抑制和诱导凋亡作用。方法采用MTT法观察Zey对PC-3细胞和正常前列腺基质细胞WPMY-1的增殖抑制作用,应用平板克隆形成实验观察Zey对PC-3细胞克隆形成的作用,AO/EB荧光染色观察细胞形态、JC-1染色观察Zey对细胞内线粒体膜电位的影响,An-nexin V-FITC/PI双染检测凋亡细胞比率,Western blot检测凋亡相关蛋白Caspase-9、Caspase-8、Caspase-3、Caspase-7和PARP的激活及Bcl-2、Bax、Bid、Bcl-xL蛋白的表达。结果Zey可以明显抑制人前列腺癌PC-3细胞的增殖并诱导其凋亡,而对正常前列腺基质细胞WPMY-1的抑制作用显著低于PC-3细胞。Zey抑制PC-3细胞克隆形成并明显诱导其凋亡。线粒体膜电位检测结果显示Zey导致细胞内线粒体膜电位的明显降低,Western blot检测结果表明Caspase-8、Caspase-9、Caspase-7、Caspase-3被激活,PARP和Bid被剪切活化,Bcl-2、Bcl-xL表达下降,而Bax表达上调。结论 Zey可选择性抑制人非激素依赖性前列腺癌PC-3细胞的增殖并诱导其凋亡,作用机制与其对Bcl-2和Caspase家族蛋白的影响,从而诱导PC-3细胞发生内源性和外源性凋亡相关。Zey有望成为治疗前列腺癌的候选药物。Aim To investigate the inhibitory effect of Zeylenone（Zey） the proliferation and apoptosis of hu- man prostate carcinoma PC-3 cells in vitro. Methods MTT assay was used to detect the viability of PC-3 and WPMY-1 normal prostate stromal cells. The inhib- itory effect of Zey on PC-3 clonogenic survival was as- sayed by the method of plate clone formation. The ap- optosis of cell in morphology was observed by AO/EB fluorescence double-dye technology. Annexin V- FITC/PI double staining and JC-1 staining assay were used to test apoptotic ratio and mitoehondrial potential of PC-3 cells, respectively. Western blot was employed to analyze the activation of Caspase-3, -8, -9, -7 and PARP, as well as the expression of Bcl-2, Bax, Bcl- XL, and Bid. Results MTF results showed that Zey exhibited significantly higher cytotoxicity to the cancer cell line PC-3 than to noncancerous cell line WPMY-1. Zey significantly inhibited the clone formation and in-duced a dose-dependent apoptosis of PC-3 cells. Mito- chondrial membrane potential decreased significantly after Zey exposure. Western blot results showed that ex- pression of proCaspase- 9, -8,-7,-3, and PARP was decreased after Zey treatment, while cleaved Caspase- 9, -8, -7, -3 and cleaved PARP increased. At the same time, the expression of Bcl-2, Bcl-xL decreased and Bax increased, and the BH3-only protein Bid was cleaved into a truncated Bid. Conclusion Zey pos- sesses potent tumor selected toxicity and can induce apoptosis of PC-3 cells through mitochondria- and Fas- mediated Caspase-dependent pathways, which is asso- ciated with Bcl-2 protein family. Thus, Zey may have great potential in the prevention and treatment of hu- man prostate tumors.

关 键 词：Zeylenone Uvaria grandiflora Roxb 人前列腺癌 PC-3细胞 细胞凋亡 Caspase活化 Bcl-2蛋白家族 

分 类 号：R284.1[医药卫生—中药学] R329.24[医药卫生—中医学]