冬凌草甲素抑制人卵巢癌细胞恶性行为及机制研究  

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

作　　者：刘艺[1] 郭金兴 LIU Yi;GUO Jin-xing(College of Pharmacy,Mudanjiang Medical University,Mudanjiang,Heilongjiang 157011,China)

机构地区：[1]牡丹江医学院药学院,黑龙江157011 [2]牡丹江医学院附属红旗医院

出　　处：《现代预防医学》2021年第1期139-143,共5页Modern Preventive Medicine

基　　金：黑龙江省卫生计生委科研课题(2018M03031)。

摘　　要：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、80μmol/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7.29μmol/L。与对照组比较,5、10和20μmol/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20μmol/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20μmol/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。Objective To 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(IC50)in SKOV3 cells after treatment with different concentrations(0,5,10,20,40 and 80μmol/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)andβ-catenin,C-myc and Cyclin D1.Results The oridonin inhibited SKOV3 cell viability in a time-dose-dependent manner,and IC50 were 23.57,12.48 and 7.29μmol/L after 24,48 and 72h treatment.After treatment with SKOV3 cells at a concentration of 5,10 and 20μmol/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 ofβ-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.

关 键 词：冬凌草甲素 卵巢癌 迁移 侵袭 WNT/Β-CATENIN信号通路 

分 类 号：R737.31[医药卫生—肿瘤]