摘要
目的:结合体内外实验研究南蛇藤提取物对人肝癌细胞Hep G2上皮间质转化的作用及分子机制。方法:取对数生长期Hep G2细胞,除对照组外,分别给予南蛇藤提取物10、20、40、80、160μg·m L-1培养,采用Western Blotting法检测上皮间质转化(EMT)相关蛋白的表达水平;斑马鱼胚胎,于受精后48 h开始给药,正常组给予E3缓冲液,DMSO组为含1%DMSO的E3缓冲液,南蛇藤组各药物浓度分别为10、20、40、80、160μg·m L-1;给药24 h后,以Western Blotting法检测m TOR信号通路相关蛋白的表达水平。结果:与对照组相比,南蛇藤提取物各浓度组明显增加E-cadherin的表达量,降低vimentin和m TOR信号通路相关蛋白的表达水平。结论:南蛇藤提取物能在一定程度上逆转人肝癌细胞EMT,其分子机制可能与m TOR信号通路相关,m TOR可作为临床治疗肝癌的新靶点。
This study was aimed to investigate the effect of Nan-She-Teng （Celastrus orbiculatus） extract on epithe-lial-mesenchymal transition（EMT） in HepG2 cells. Except the control group, human hepatocellular carcinoma HepG2 cells in other groups were treated with Celastrus Orbiculatus extract in different concentrations （10, 20, 40, 80, and 160 μg·mL-1）. The protein expression levels related to EMT were detected by western blotting. At 48 h after fertiliza-tion, the zebrafish embryos were randomly assigned to 7 groups as follows： untreated control group （E3 buffer）, DMSO group （E3 buffer with 1% DMSO）, and different dosages treatment of C.orbiculatus extract （10, 20, 40, 80, and 160μg·mL-1） for 24 h. The protein expressions of mTOR signaling pathways were detected by western blotting. The re-sults showed that compared with the control group, C.orbiculatus extract significantly increased E-cadherin protein expression. Simultaneously, C.orbiculatus extract inhibited vimentin and mTOR signaling pathways at protein levels. It was concluded that to a certain extent, C.orbiculatus extract prevented EMT in HepG2 cells by modulating the mTOR signaling pathway. Therefore, it suggested that mTOR can be chosen as a new therapeutic target for clinical treatment of hepatic carcinoma.
出处
《世界科学技术-中医药现代化》
北大核心
2014年第12期2647-2651,共5页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
国家自然科学基金委青年基金(81403232):南蛇藤提取物靶向PI3K/Akt/mTOR信号通路抑制肝癌早期转移的作用及机制研究
负责人:钱亚云
国家自然科学基金委面上项目(81274141):南蛇藤总萜通过协同抑制Notch和VEGF的表达阻断肝癌血管生成的作用机制
负责人:刘延庆
江苏省自然科学基金委面上项目(BK2012686):南蛇藤提取物增强抑癌基因MASPIN表达抑制胃癌转移的机制研究
负责人:钱亚云
教育部博士点基金新教师类项目(20133250120003):南蛇藤总萜靶向mTOR信号通路抑制肝癌转移的分子机制研究
负责人:钱亚云
教育部大学生科创项目(201311117042):南蛇藤总萜靶向mTOR信号通路抑制肝癌转移的分子机制研究
负责人:吉兰芳
姜馨
江苏省教育厅大学生科创重点项目(201311117042z):南蛇藤总萜靶向mTOR信号通路抑制肝癌转移的分子机制研究
负责人:吉兰芳
姜馨
关键词
南蛇藤
肝癌
上皮间质转化
斑马鱼
信号通路
Nan-She-Teng （Celastrus orbiculatus）, hepatic carcinoma, epithelial-mesenchymal transition,zebrafish, signaling pathway
