冬凌草甲素抑制人卵巢癌细胞恶性行为及机制研究

目的 研究冬凌草甲素（Oridonin）对人卵巢癌（Human ovarian cancer）SKOV3 细胞迁移和侵袭能力的影响及其潜在的分子机制。方法 采用CCK-8 法检测不同浓度（0、5、10、20、40、80μmol/L）的冬凌草甲素作用SKOV3细胞24、48 和72h 后细胞活力的变化，计算半数抑制浓度（IC50）。采用Annexin V-FITC/PI 双染法检测SKOV3细胞凋亡。采用划痕修复实验检测SKOV3细胞的迁移能力，Transwell小室实验检测SKOV3的侵袭能力。Western blot检测SKOV3细胞上皮细胞间充质转化（EMT）蛋白［E-钙黏蛋白（E-cadherin）、波形蛋白（Vimentin）］和Wnt/β-连环蛋白（β-catenin）信号通路中β-catenin及下游靶分子原癌基因（C-myc）、细胞周期蛋白D1（Cyclin D1）蛋白的表达。结果 冬凌草甲素可抑制SKOV3 细胞活力，且具有时间-剂量依赖性，作用24、48 和72 h后IC50值为23.57、12.48和7.29μmol/L。与对照组比较，5、10和20μmol/L冬凌草甲素作用SKOV3细胞24h 后，细胞凋亡率明显升高，迁移率和侵袭率降低（P<0.05）。与对照组比较，5、10和20μmol/L冬凌草甲素可升高E-cadherin 蛋白表达（P<0.05），降低Vimentin 表达（P<0.05）。与对照组比较，5、10和20 μmol/L冬凌草甲素可降低β-catenin、C-myc和Cyclin D1表达（P<0.05）。结论 冬凌草甲素具有抑制SKOV3细胞增殖、转移和侵袭能力的作用，该作用与其抑制Wnt/β-catenin 信号通路有关。

Effect and mechanism of oridonin on malignant behavior of human ovarian cancer

?o study the effects of oridonin on the migration and invasion of human ovarian cancer SKOV3 cells and its potential molecular mechanism.Methods The CCK-8 assay was used to detect the changes of cell viability and the half-inhibitory concentration (IC50) in SKOV3 cells after treatment with different concentrations (0, 5, 10, 20, 40 and 80μmol/L) of the oridonin for 24, 48, and 72h. The Annexin V-FITC/PI double staining method was used to detect the the apoptosis. The wound healing and the Transwell assay were conducted to detect migration and invasion, respectively. The Western blot was used to detect epithelial-mesenchymal transition (EMT) proteins (E-cadherin, Vimentin) and β-catenin, C-myc and Cyclin D1. Results The oridonin inhibited SKOV3 cell viability in a time-dose-dependent manner, and IC50 were 23.57, 12.48 and 7.29μmol/L after 24, 48 and 72h treatment. After treatment with SKOV3 cells at a concentration of 5, 10 and 20μmol/L oridonin for 24 h, SKOV3 cells were obtained. Compared with the control group, the oridonin increased significantly apoptosis rate and decreased the migration rate and the invasion rate (P<0.05). Compared with the control group, the oridonin concentration-dependently increased the expression of E-cadherin (P<0.05) and decreased the expression of Vimentin (P<0.05). Compared with the control group, the oridonin concentration-dependently reduced the expressions of β-catenin, C-myc and Cyclin D1 (P<0.05). Conclusion The oridonin can inhibit the proliferation, the migration and invasion of SKOV3 cells, which is related to the down-regulation of Wnt/β-catenin signaling pathway.