摘要
目的观察冬凌草甲素对人宫颈癌HeLa细胞增殖、凋亡、迁移、侵袭及Wnt/β-连环蛋白(β-catenin)信号通路的影响。方法体外培养HeLa细胞,以细胞计数试剂盒-8(cell counting kit-8,CCK-8)检测不同浓度(0、10、20、40、80、160μmol·L^(-1))的冬凌草甲素对HeLa细胞处理不同时间(12 h、24 h和36 h)的细胞活力的影响。以0(对照组)、10、20、40μmol·L^(-1)冬凌草甲素处理HeLa细胞24 h,TUNEL染色和AnnexinV-FITC/PI双染法分别检测HeLa细胞凋亡,细胞粘附实验检测HeLa细胞粘附力,划痕修复实验检测HeLa细胞迁移能力,Transwell小室体外侵袭实验检测HeLa细胞侵袭能力,Western blot分析检测Wnt/β-catenin信号通路中β-catenin及下游靶分子原癌基因(C-myc)、细胞周期素D1(Cyclin D1)蛋白的表达。结果冬凌草甲素以浓度-时间依赖效应的方式抑制HeLa细胞细胞活力(P<0.05)。与对照组比较,10、20、40μmol·L^(-1)冬凌草甲素组显著升高HeLa细胞凋亡率和凋亡指数(P<0.05);与对照组比较,10、20、40μmol·L^(-1)冬凌草甲素组显著降低HeLa细胞黏附率、迁移率和侵袭率(P<0.05);与对照组比较,10、20、40μmol·L^(-1)冬凌草甲素组显著降低HeLa细胞β-catenin、C-myc和Cyclin D1蛋白表达(P<0.05)。结论冬凌草甲素具有抑制HeLa细胞增殖的作用,亦可诱导凋亡及抑制迁移和侵袭能力,并抑制Wnt/β-catenin信号通路。
Objective To observe effects of oridonin on proliferation,apoptosis,migration,invasion and the Wnt/β-catenin signaling pathway of human cervical carcinoma HeLa cells.Methods HeLa cells were cultured in vitro and treated with different concentrations(0,10,20,40,80 and 160μmol·L^(-1))of oridonin for different time(12 h,24 h and36 h),and effects on cell viability were detected by cell counting kit-8(CCK-8).The HeLa cells were treated with 0(the control group),10,20 and 40μmol·L^(-1) of oridonin for 24 h.The HeLa cell apoptosis was detected by TUNEL staining and Annexin V-FITC/PI double staining,respectively.The HeLa cell adhesion was detected by cell adhesion assay.The HeLa cell migration and invasion was detected by wound healing assay and in vitro Transwell chamber invasion assay,respectively.Protein expressions ofβ-catenin in the Wnt/β-catenin signaling pathway,downstream target molecule Cmyc and Cyclin D1 were detected by Western blot analysis.Results Oridonin inhibited the cell viability in a concentration-time-dependent effect manner(P<0.05).Compared with the control group,10,20 and 40μmol·L^(-1) oridonin significantly increased the apoptotic ratio and the apoptotic index of the HeLa cells(P<0.05).Compared with the control group,10,20 and 40μmol·L^(-1) oridonin significantly decreased the adhesion ratio,migration ratio and invasion ratio of the HeLa cells(P<0.05).Compared with the control group,10,20 and 40μmol·L^(-1) oridonin significantly decreased the protein expressions ofβ-catenin,C-myc and Cyclin D1 of the HeLa cells(P<0.05).Conclusion Oridonin has an effect of inhibiting proliferation and can induce apoptosis and inhibit migration,invasion and the Wnt/β-catenin signaling pathway in the HeLa cells.
作者
刘艺
郭金兴
Liu Yi;Guo Jinxing(School of Pharmacy,Mudanjiang Medical University,Mudanjiang 157011,China;The Hongqi Hospital Affiliated to Mudanjiang Medical University,Mudanjiang 157011,China)
出处
《世界科学技术-中医药现代化》
CSCD
北大核心
2020年第9期3129-3135,共7页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
黑龙江省中医药管理局科研项目(ZHY18-W39):冬凌草甲素抑制人宫颈癌HeLa细胞迁移和侵袭的可能机制研究,负责人:郭金兴。
