摘要
目的:探讨熊果酸对前列腺癌细胞系PC-3细胞凋亡的影响及其机制。方法:体外培养PC-3细胞,用不同浓度的熊果酸处理24、48、72 h,MTT法测定其对PC-3细胞增殖的影响;倒置显微镜下观察细胞的形态学变化;熊果酸(浓度分别为20、40、80μmol/L)处理PC-3细胞48 h后,分别采用流式细胞仪检测细胞凋亡,分光光度计检测Casepase-3、Casepase-9相对活性,western blot法检测COX-2、PTEN基因表达情况。结果:熊果酸(浓度10~100μmol/L)能抑制PC-3细胞的增殖,呈剂量和时间依赖性;倒置显微镜下可观察到典型的细胞凋亡形态;熊果酸(浓度分别为20、40、80μmol/L)作用PC-3细胞48 h后,细胞凋亡率分别为12.5%、23.3%、35.4%,对照组凋亡率为3.4%;Casepase-3、Casepase-9的相对活性显著增加;COX-2的表达降低,而PTEN的表达升高,呈剂量依赖性。结论:熊果酸能抑制PC-3细胞的增殖并诱导其凋亡,其作用机制可能与下调COX-2基因的表达及上调PETN基因并增加细胞Caspase-3、Caspase-9激活线粒体凋亡途径有关。熊果酸是一种前景广阔的抗肿瘤药物。
Objective : To explore the effect of Ursolic acid on cell apoptosis in PC-3 cells and discuss its possible mechanisms. Methods : PC-3 cells were cultured in vitro. After treatment by Ursolic acid at different concentrations respectively at different time points, the cell survival was determined by the MTT method. The changes of cell morphology were observed by inverted microscope. Apoptosis was detected by flow cytometry. The relative activity of Caspase-3 and Caspase-9 was monitored by spectrophotometer. The changes of protein expression of COX-2 and PTEN were detected by western blot. Results : From the data of MTT, the cell proliferation of PC-3 cells was inhibited by Ursolic acid in a dose-dependent and time- dependent manner. Typical apoptosis morphology of PC-3 cells was observed by inverted microscope. Flow cytometry assays showed that Ursolic acid significantly induced apoptosis in PC-3 cells. After treated with Ursolic acid, the apoptosis rate of PC-3 cells was 12.5%,23.3%, and 35.4% respectively, which showed an obvious concentration-effect relationship. The relative activity of Caspase-3 of Ursolic acid group was 0.194 ±0.02,0.253 ±0.04 and 0.316 ±0.03 respectively. And the relative activity of Caspase-9 was 0.177 ±0.02,0.234 ±0.03 and 0.305 ±0.04. The data of western blot showed that Ursolic acid down-regulated COX-2 and up-regulated PTEN in a dose-dependent manner. Conclusion: Ursolic acid can inhibit the proliferation of PC-3 cells and induce apoptosis, and the mechanism of Ursolic acid on apoptosis may be related to the down regulation of COX-2 expression and up regulation of PTEN expression, as well as the increase of relative activity of Caspase-3 and Caspase-9. Ursolic acid may be a promising anti-tumor agent for mammary cancer treatment.
出处
《辽宁中医药大学学报》
CAS
2014年第7期37-40,共4页
Journal of Liaoning University of Traditional Chinese Medicine