环王巴明诱导DU145细胞凋亡的机制研究

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

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-2 mRNA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-2mRNA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-2mRNA表达,活化细胞凋亡的线粒体途径有关。

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.