目的：探讨五味子乙素（ Sch B）对人卵巢癌Skov3细胞株的增殖、凋亡及Wnt/β-catenin信号通路的影响。方法采用0、1�0、10�0、20�0、50�0μmol/L Sch B处理Skov3细胞，采用四甲基偶氮唑盐（ MTT）法检测各浓度处理24、48、72和96h的增殖抑制率，Annexin-FITC/PI双染法检测不同浓度处理48h后的细胞凋亡情况，流式细胞仪检测各浓度处理48h后的细胞周期分布情况，Western blotting检测各浓度处理48h后细胞核Wnt/β-catenin信号通路中β-连接素（β-catenin）及下游靶分子C-myc、细胞周期素D1（ Cyclin D1）的蛋白水平，同时于各浓度处理48h后检测细胞质和细胞核的糖原合成酶激酶-3β（ GSK-3β）活性。结果 Sch B可呈剂量和时间依赖方式增加细胞增殖抑制率（ P＜0�05），作用48h后可呈剂量依赖方式升高早、晚期凋亡率及细胞质/胞核 GSK-3β活性（ P＜0�05）；除1�0μmol/L外，其余浓度作用48h 的 G0/G1期细胞比例均高于0μmol/L，S 期、G2/M 期细胞比例及β-catenin、C-myc 和 Cyclin D1蛋白水平均低于0μmol/L （ P＜0�05），且10�0、20�0、50�0μmol/L Sch B间的差异有统计学意义（ P＜0�05）。结论 Sch B可以抑制Skov3细胞株的增殖并促进其凋亡和细胞周期阻滞，以及抑制Wnt/β-catenin通路的激活。%Objective To explore the influences of Schisandrin B ( Sch B) on proliferation, apoptosis and Wnt/β-catenin signaling pathway of SKOV3 human ovarian cancer cells. Methods The SKOV3 cells were treated with different concentrations of Sch B ( 0, 1�0, 10�0, 20�0, 50�0μmol/L) . The MTT was used to measure the proliferation inhibition rates at 24, 48, 72 and 96h treated with different concentrations of Sch B. The Annexin-FITC/PI double staining was employed to detect cell apoptosis at 48h after treat-ment with different concentrations of Sch B. The cycle distribution at 48h after treatment with Sch B was detected by flow cytometry. The Western blotting was used to measure the nuclearβ-catenin protein levels in Wnt/β-catenin signaling pathway as well as its downstream target C-myc and cyclin D1. The cytoplasmucleus activities of glycogen synthase kinase 3β ( GSK-3β) at 48h after treatment with different concentrations of Sch B were measured by activity assay kit. Results The Sch B can increase the proliferation inhibition rates in a dose-and time-dependent manner, and elevate the early and late apoptosis and cytoplasmucleus activities of GSK-3βat 48h after treatment. In addition to 1�0μmol/L, the proportion of cells in G0/G1 phase of the remaining concentrations were higher than those of 0μmo/L ( P<0�05) , and the proportion of cells in S phase and G2/M phase andβ-catenin, C-myc and Cyclin D1 protein levels were lower than those of 0μmo/L ( P<0�05) . Conclusion Sch B can inhibit the proliferation, promote apoptosis and cell cycle arrest and inhibit the activation of Wnt/β-catenin pathway.
展开▼
