摘要
探讨圆齿野鸦椿中野鸦椿酸(EA)对人肝癌细胞增殖、侵袭和迁移能力的影响及其机制研究。从圆齿野鸦椿果皮中提取得到EA,通过培养人肝癌细胞Hep G2细胞,以MTT法分析EA对Hep G2细胞增殖的影响,流式细胞术评价不同浓度(20、40、80μmol/L)的EA对Hep G2细胞的凋亡状态和周期分布情况,细胞划痕、Transwell小室实验考察对Hep G2细胞侵袭转移能力的变化,用Western blot法和荧光定量PCR法检测野鸦椿酸对上皮-间质转化(EMT)相关标志物E-Cadherin、N-Cadherin、Vimentin、MMP-2和MMP-9的蛋白和mRNA表达的影响。结果显示,EA抑制人肝癌细胞Hep G2细胞的增殖,24、48、72 h的IC50分别为32. 16±4. 58、26. 45±3. 79、和16. 76±4. 01μmol/L。随EA浓度的增大,细胞的凋亡率逐渐升高,且发生显著的G0/G1期阻滞。EA可降低Hep G2细胞的侵袭和迁移能力,上调E-Cadherin的蛋白和mRNA的水平,下调N-Cadherin、Vimentin、MMP-2和MMP-9的蛋白和mRNA表达。EA抑制人肝癌细胞增殖和侵袭转移能力,可能与其调控EMT相关信号通路有关。
The purpose of the present work was to study the inhibitory effect of Euscaphic acid(EA)on the proliferation,invasion and migration of human hepatocarcinoma HepG2 cells.EA was extracted from the pericarps of Euscaphis konishii Hayata.MTT assay was employed to evaluate the proliferative inhibition of EA on HepG2 cells;flow cytometry was adopted to test cell cycle and apoptosis status;wound healing and transwell assays were performed to determine the effect of EA on invasion and migration abilities.The protein and mRNA expression of E-Cadherin,N-Cadherin,Vimentin,MMP-2 and MMP-9 were evaluated using Western blot and qRT-PCR.The results showed that EA significantly inhibits the proliferation of HepG2,with the IC 50 of 32.16±4.58,26.45±3.79 and 16.76±4.01μmol/L for 24,48 and 72 h treatment,respectively.EA induced apoptosis and G0/G1 phase arrest in a dose-dependent manner,suppressed invasion and migration ability of HepG2,and mediated the Epithelial-mesenchymal transition(EMT)process of HepG2 cells via upregulating the protein and mRNA levels of E-Cadherin and downregulating the protein and mRNA expression of N-Cadherin,Vimentin,MMP-2 and MMP-9.In summary,the inhibition of proliferation,invasion and migration of human heptocarcinoma HepG2 cells by EA probably through the EMT associated signaling pathway.
作者
黄维
冯翯
郑毅
张泰雷
倪林
姚霞
邹小兴
HUANG Wei;FENG He;ZHENG Yi;ZHANG Tai-lei;NI Lin;YAO Xia;ZOU Xiao-xing(College of Life Sciences,Fujian Agriculture and Forestry University;Engineering Research Institute of Conservation,Utilization of Natural Bioresources,Fujian Agriculture and Forestry University;College of Forestry,Fujian Agriculture and Forestry University;College of Plant Protection,Fujian Agriculture and Forestry University,Fuzhou 350002,China;China National of Traditional&Herbal Medicine Co.,Ltd.,Beijing 100095,China)
出处
《天然产物研究与开发》
CAS
CSCD
北大核心
2018年第9期1502-1508,共7页
Natural Product Research and Development
基金
国家自然科学基金(31700292)
福建省自然科学基金(2018J01653)
中国博士后科学基金(2018M632566)
福建农林大学科技创新专项(CXZX2016072)
关键词
野鸦椿酸
圆齿野鸦椿
增殖
侵袭
迁移
euscaphic acid
Euscaphis konishii Hayata
proliferation
invasion
migration
