Zeylenone抑制人前列腺癌PC-3细胞增殖及诱导凋亡作用研究

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。

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

Aim To investigate the inhibitory effect of Zeylenone(Zey) the proliferation and apoptosis of human 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 inhibitory effect of Zey on PC-3 clonogenic survival was assayed by the method of plate clone formation.The apoptosis 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 mitochondrial 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 MTT 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 induced a dose-dependent apoptosis of PC-3 cells.Mitochondrial membrane potential decreased significantly after Zey exposure.Western blot results showed that expression 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 possesses potent tumor selected toxicity and can induce apoptosis of PC-3 cells through mitochondria-and Fas-mediated Caspase-dependent pathways,which is associated with Bcl-2 protein family.Thus,Zey may have great potential in the prevention and treatment of human prostate tumors.