摘要
茶氟香酰胺(Ethyl 6-fluorocoumarin-3-carboxylyl L-theanine,TFC)是对茶氨酸进行结构优化后的产物,通过实验研究其对人肝癌细胞(SMMC7721)生长和迁移的抑制作用,并了解其作用机制.MTT和Transwell chamber法分别检测不同浓度的TFC对SMMC7721细胞生长和迁移的影响,流式细胞术分析不同浓度的TFC对SMMC7721细胞凋亡的影响,Western blotting检测不同浓度的TFC组对与SMMC7721细胞中与肿瘤细胞生长、迁移和凋亡密切相关蛋白表达的影响.实验结果显示,TFC能够显著抑制SMMC7721细胞生长、迁移并诱导癌细胞凋亡,TFC药物分子作用机制可能与下调SMMC7721细胞中血管内皮生长因子受体VEGFR1、磷酸化蛋白激酶B(phosphorylated protein kinase B,p AKT)、核转录因子NF-κB、抗凋亡蛋白(BCL2,apoptosis regulator,BCL2)、细胞周期蛋白(cyclin D1,CCND1)表达和上调促凋亡蛋白(BCL2 associated X,apoptosis regulator,BAX)、细胞色素c(cytochrome c,somatic,CYCS)、人体抑癌基因蛋白(tumor protein p53,TP53)、半胱天冬酶3(caspase,CASP3)和E-cadherin蛋白表达有关,TFC抑制人肝癌SMMC7721细胞生长和迁移作用的重要信号通路涉及到VEGFR/AKT/NF-κB.
The purpose of this study is to investigate the inhibitory effects of TFC,a synthesized theanine derivative on the growth and migration of human hepatoma cells.The assays of MTT and transwell chamber are applied to evaluate the growth and migration of the human hepatoma cells treated with different concentrations of TFC,respectively.Flow cytometry is used to detect the TFC-induced apoptosis in the hepatoma cells stained with AnnexinV-FITC and propidium iodide(PI).Western blot is used to analyze the related protein expression in the hepatoma cells.TFC significantly inhibites the growth and migration of the hematoma cells in a dose-time dependent manner.TFC induces the apoptosis in the hepatoma cells.The molecular mechanisms of TFC action may be related to the reduction of the protein levels in phosphorylated protein kinase B(pAKT),vascular endothelial growth factor receptor(VEGFR),nuclear factor kappa B(NF-κB),BCL2,apoptosis regulator(BCL2),cyclin D1(CCND1)and to the increase in the protein expressions of BCL2 associated X,apoptosis regulator(BAX),TP53,caspase 3(CASP3),cytochrome c(CYCS)and E-cadherin.Inhibition of the hematoma cell growth and migration by TFC is involved in suppression of the important signaling pathway of VEGFR/AKT/NF-κB.
作者
郑晓慧
朱荣芹
刘真真
李筝
盖静
杨美玲
闫春燕
刘昆
张国营
ZHENG Xiao-hui;ZHU Rong-qin;LIU Zhen-zhen;LI Zheng;GE Jing;YANG Mei-ling;YAN Chun-yan;LIU Kun;ZHANG Guo-ying(Laboratory of Molecular Pharmacology,School of Pharmacy,Yantai University,Collaborative Innovation Center of Advanced Drug Delivery System and Biotech Drugs in Universities of Shandong,Key Laboratory of Molecular Pharmacology and Drug Evaluation(Yantai University),Ministry of Education,Yantai Uni)
出处
《烟台大学学报（自然科学与工程版）》
CAS
2018年第2期136-144,共9页
Journal of Yantai University(Natural Science and Engineering Edition)
基金
山东省自然科学基金资助项目(ZR2015HM004)
