鱼藤素诱导人前列腺癌细胞凋亡及其机制研究

Studies on the mechanism of Deguelin-induced apoptosis of human prostate cancer cell lines

目的观察鱼藤素对于激素抵抗型前列腺癌（HRPC）细胞PC3和DU145增殖、细胞周期和凋亡的影响并探讨其机制。方法设阴性对照组（有细胞但不加药），空白对照组（无细胞仅有培养液），阳性对照组（渥曼青霉素100nmol／L），及鱼藤素分别10、100、1μmol／L共6组。CCK-8法进行细胞毒性实验，检测细胞生长抑制率。流式细胞术检测细胞周期和凋亡，Western-blot检测Akt、MAPK及其磷酸化蛋白表达，探讨药物作用机制。结果10nmol／L～1μmol／L鱼藤素对PC3细胞均有生长抑制作用，呈现明显的时间、浓度依赖性，对DU145细胞则无此作用。鱼藤素使PC3细胞出现G2／M期阻滞现象并引起浓度依赖性的凋亡，而未改变DU145细胞的周期分布也不能诱导其凋亡。鱼藤素能够阻断P13K／AKT通路而对MAPK通路无影响。结论鱼藤素通过阻断P13K／AKT通路实现抑制PC3细胞增殖、诱导凋亡的作用。两株细胞问实验结果的差异是因为其P13K／AKT通路活化状态的差异造成的。

Objective To investigate the anticancer effects and molecular mechanism of deguelin on human prostate cancer cells PC3 and DU145 in vitro. Methods Ceils were treated with either 0.1% DMSO as diluent control, wortmannin 100 nmol/L as positive control or various concentrations of deguelin （ 10 nmol/L, 100 nmol/L and 1 μmol/L）. CCK-8 assay was used to identify the inhibitory effect of Deguelin on proliferation in PC3 and DU145 cells. Apoptosis was assessed through annexin V/PI double-labeled eytometry. The effect of deguelin on the cell cycle of PC3 and DU145 cells was studied by propidium iodide method. Western blot assay was used to detect the expression of AKT, p42/44 MAPK and their phosphorylated proteins. Results Deguelin 10 nmol/L -1μmoL/L could inhibit the proliferation of PC3 cells but had little effect on DUI45 cells. Deguelin significantly increased the percentage of G2/M phase and induced apoptosis of PC3 cells ,but didn＇ t alter cell cycle and induce apoptosis of DU145 ceils. The expression of pAKT protein was decreased in PC3 cells treated with Deguelin but not in DU145 cells. Conclusion Deguelin can inhibit cell growth, induce G2/M arrest and apoptosis of PC3 cells by blocking PI3K/AKT pathway but has little effect on DU145 ceils. The different effects of Deguelin on PC3 and DU145 cells are caused by the different activated status of PI3K/AKT pathway between the two cells.