环王巴明诱导DU145细胞凋亡的机制研究

目的：研究环王巴明诱导DU145细胞凋亡作用及其对Gli-1和Bcl-2 mRNA表达的影响，探讨环王巴明诱导DU145细胞凋亡的机制。方法：不同浓度环王巴明处理DU145细胞后，吖啶橙染色，荧光显微镜下观察细胞形态变化。流式细胞术观察细胞凋亡率变化，RT—PCR检测5、10μmol／L环王巴明作用72h后的实验组和对照组Gli-1、Bcl-2mRNA表达水平差异。结果：当环王巴明浓度大于5μmol／L作用72h后，荧光显微镜下可见细胞核固缩或碎裂，核仁变形等典型的凋亡形态学改变。环王巴明5μmol／L组Gli-1、Bcl-2 mRNA表达减弱，但同对照组差异无统计学意义（P〉0．05），与空白及DMSO对照组相比10μmol／L环王巴明可以显著下调Gli—1、Bcl-2 mRNA表达（P〈0．01）。结论：环王巴明可诱导人前列腺癌DU145细胞株凋亡，其作用机制可能与下调Gli-1和Bcl-2 mRNA表达，活化细胞凋亡的线粒体途径有关。

Mechanism of cyclopamine induced apoptosis in human prostate cancer DU145 cell lines

Objective：To investigate the effect of cyclopamine induce apoptosis to human prostate cancer cell line DUI45 ,and its impact on Gli - land Bcl -2 mRNA levels. Methods：To treat Du145 cells with cyclopamine at different concentration for 72h, and observe the morphology. Gli -1 and Bcl- 2 mRNA expression was examined by RT- PCR, and westemblot. Results ： After cyclopamine of 5μmol/L treated for 72h, the nucleus pycnosis, fragmentation, chromatospherite disfiguration apoptotic morphology were observed in DU145 cells under fluorescence microscope. The expression levels of Gli - 1 and Bcl -2 mRNA in 5 μmol/L cyclopamine group were decreased but no statistical significance（ P 〉0.05 ） ,While 10μmol/L cyclopamine group was down - regulated significantly compare with control and DMSO group（ P 〈 0.01 ）. Conclusion：The mechanism of cyclopamine induced apoptosis in DUI45 cell lines may be related to down- regulation of expression levels of Gli- 1 and Bcl-2 mRNA,and active the mitochondrial apoptotic pathways.