南蛇藤提取物靶向mTOR抑制人肝癌HepG2细胞侵袭与转移能力的研究  

Celastrus orbiculatus extracts inhibit invasion and metastasis in human hepatocellular carcinoma HepG2 cells by targeting mTOR

作　　者：钱亚云[1] 李文原 赵雪煜 杨婷[1] 严妍[1] 房传赐 侯晶晶 刘延庆[1] QIAN Ya-yun;LI Wen-yuan;ZHAO Xue-yu;YANG Ting;YAN Yan;FANG Chuan-ci;HOU Jing-jing;LIU Yan-qing(Medical College,Yangzhou University,Yangzhou 225001,China)

机构地区：[1]扬州大学医学院,江苏扬州225001

出　　处：《中草药》2018年第20期4831-4837,共7页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金资助项目(81403232;81573656);江苏省自然科学基金项目(BK20171290;BK2012686);教育部博士点基金新教师类项目(20133250120003)

摘　　要：目的探索南蛇藤提取物(COE)靶向哺乳动物雷帕霉素靶蛋白(m TOR)抑制人肝癌Hep G2细胞增殖、侵袭与转移的分子机制。方法用si RNA技术,构建敲除m TOR基因的Hep G2细胞模型,MTT法检测COE对Hep G2/m TOR-细胞增殖能力的影响。划痕实验及Transwell实验检测药物对细胞侵袭转移能力的影响。Western blotting法分析用药后基质金属蛋白酶-2(MMP-2)、MMP-9蛋白的表达水平变化。结果成功构建敲除m TOR表达的Hep G2/m TOR-细胞。COE明显抑制Hep G2/m TOR-细胞的增殖(P<0.05),并呈浓度依赖性。划痕实验结果显示COE降低了细胞迁移能力。Transwell实验结果表明,COE(80 mg/L)明显减少了穿膜细胞数目(P<0.05)。Western blotting结果显示,COE(80 mg/L)明显降低MMP-2、MMP-9蛋白的表达水平(P<0.05)。结论 COE明显抑制Hep G2/m TOR-细胞的增殖与侵袭转移,m TOR通路可能是其抑制肝癌的潜在作用靶点之一。Objective To investigate the molecular mechanisms of Celastrus orbiculatus extracts（COE） of the invasion and metastasis inhibition in human hepatocellular carcinoma Hep G2 cells by targeting m TOR.Methods The Hep G2/m TOR-cells with m TOR knockout expression were constructed by using si RNA technology.The effect of COE on the proliferation of the Hep G2/m TOR-cells was also studied.The Hep G2/m TOR-cells were treated with COE in different concentrations（20,40,80,160,and 320 mg/L） for 24 h.The cell reproductive capability of Hep G2/m TOR-cells was detected by MTT.The effect of COE on the metastatic ability of Hep G2/m TOR-cells in vitro was investigated by scratch assay and Transwell migration assay.The expression levels of molecular mechanisms related proteins MMP-2 and MMP-9 were assessed by Western blotting.Results The Hep G2/m TOR-cells with m TOR knockout expression were successfully constructed.COE significantly inhibited the proliferation of Hep G2/m TOR-cells in a concentration-dependent manner（P 0.05）.COE decreased the invasion and migration of Hep G2/m TOR-cells.The results of Transwell experiment indicated that COE（80 mg/L） significantly reduced the number of transmembrane cells（P 0.05）.And the expression levels of MMP2 and MMP9 protein were significantly reduced in the Hep G2/m TOR-cells after the treatment of COE.Conclusion COE can significantly inhibit the proliferation,invasion,and migration in the Hep G2/m TOR-cells.Our data reveal that COE is a potential chemotherapeutic drug in human hepatocellular carcinoma treatments via targeting m TOR signal pathway.

关 键 词：南蛇藤 原发性肝细胞癌 哺乳动物雷帕霉素靶蛋白 侵袭 转移 

分 类 号：R285.5[医药卫生—中药学]