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目的:研究桦木醇对人卵巢癌细胞CAOV3的细胞增殖和凋亡的影响并探讨其作用机制。方法:四甲基偶氮唑盐(MTT)比色法测定不同浓度的桦木醇对CAOV3细胞生长的抑制效果,DAPI染色后观察细胞产生凋亡的形态学变化,流式细胞技术分析细胞凋亡,体外caspase活力测定凋亡相关蛋白caspase-3/-8/-9的激活情况,蛋白质免疫印迹技术检测caspase-3底物多聚(ADP-核糖)聚合酶[Poly(ADP-ribose)polymerase,PARP]断裂的情况。结果:桦木醇可有效地抑制CAOV3细胞的增殖并呈量-效关系,IC50值为8μg/mL;DAPI染色呈现典型的细胞凋亡特征,流式结果显示12 h后细胞凋亡率明显增加,caspase-3/-9活性显著上升,免疫印迹检测结果显示PARP断裂。结论:桦木醇抑制卵巢癌细胞CAOV3的增殖并诱导caspase参与的细胞凋亡,并且这种细胞凋亡涉及线粒体通路。
Objective: To investigate the effect of betulin on the proliferation and apoptosis of human ovarian cancer cell line CAOV3 and to explore its mechanism. Methods: MTT assay was used to determine the inhibitory effect of different concentrations of betulin on the growth of CAOV3 cells. The apoptosis of CAOV3 cells was observed by DAPI staining. The apoptosis of CAOV3 cells was analyzed by flow cytometry (ADP-ribose) polymerase (Poly (ADP-ribose) polymerase) was detected by Western blotting, and the activation of caspase-3 / -8 / PARP] broken. Results: Betulin could effectively inhibit the proliferation of CAOV3 cells in a dose-effect relationship with an IC50 value of 8μg / mL. DAPI staining showed typical characteristics of apoptosis. Flow cytometry results showed that apoptosis rate of CAOV3 cells increased significantly after 12 hours, The activity of caspase-3 / -9 increased significantly, and the result of immunoblot showed that PARP was cleaved. CONCLUSION: Betulin inhibits the proliferation of ovarian cancer cell line CAOV3 and induces caspase-associated apoptosis, and this apoptosis involves mitochondrial pathway.