异钩藤碱调节EGFR/FAK信号通路对肝癌细胞增殖、迁移和5-FU耐药性的影响  

Effects of isorhynchophylline on proliferation,migration,and 5-FU resistance of liver cancer cells by regulating EGFR/FAK signaling pathway

作　　者：李海燕 栗昭生 赵龙 金春花 张鼐鹏 LI Haiyan;LI Zhaosheng;ZHAO Long;JIN Chunhua;ZHANG Naipeng(Department of Pharmacy,Hongqi Hospital Affiliated to Mudanjiang Medical University,Mudanjiang 157000,China;First Clinical Medical College,Mudanjiang Medical University,Mudanjiang 157011,China;Department of Neurological Internal Medicine,Hongqi Hospital Affiliated to Mudanjiang Medical University,Mudanjiang 157000,China;Department of Second General Surgery,Hongqi Hospital Affiliated to Mudanjiang Medical University,Mudanjiang 157000,China)

机构地区：[1]牡丹江医学院附属红旗医院药学部,黑龙江牡丹江157000 [2]牡丹江医学院第一临床医学院,黑龙江牡丹江157011 [3]牡丹江医学院附属红旗医院神经内科,黑龙江牡丹江157000 [4]牡丹江医学院附属红旗医院普外二科,黑龙江牡丹江157000

出　　处：《药物评价研究》2024年第6期1267-1274,共8页Drug Evaluation Research

基　　金：黑龙江省省属高等学校基本科研业务费科研项目(2023-KYYWF-0959)。

摘　　要：目的探讨异钩藤碱调节表皮生长因子受体(EGFR)/局部黏着斑激酶(FAK)信号通路对肝癌细胞增殖、迁移和5-氟尿嘧啶(5-FU)耐药性的影响。方法取对数生长期的HepG2细胞,分为对照组,异钩藤碱低、中、高浓度(32.5、65.0、130.0μmol·L^(-1))组,异钩藤碱(130μmol·L^(-1))+EGFR激活剂NSC228155(2μmol·L^(-1))组;对数生长期的5-FU耐药肝癌细胞系HepG2/5-FU分为对照组、异钩藤碱(130μmol·L^(-1))组、5-FU(60μmol·L^(-1))组、异钩藤碱(130μmol·L^(-1))+5-FU(60μmol·L^(-1))组、异钩藤碱(130μmol·L^(-1))+5-FU(60μmol·L^(-1))+NSC228155(2μmol·L^(-1))组。给药处理24 h,对照组不给药。EdU染色、CCK-8检测HepG2或HepG2/5-FU细胞增殖;划痕实验检测HepG2或HepG2/5-FU细胞迁移;Western blotting检测细胞中细胞周期蛋白D1(CyclinD1)、迁移侵袭增强子因子1(MIEN1)、P-糖蛋白(P-gp)、p-EGFR、p-FAK蛋白表达。结果与对照组相比,异钩藤碱低、中、高浓度组EdU阳性率、吸光度(A450)值、划痕愈合率、CyclinD1、MIEN1、p-EGFR、p-FAK蛋白表达显著降低,且呈浓度相关性(P<0.05);与异钩藤碱130.0μmol·L^(-1)组相比,异钩藤碱+NSC228155组HepG2细胞EdU阳性率、A450值、划痕愈合率、CyclinD1、MIEN1、p-EGFR、p-FAK蛋白表达显著升高(P<0.05)。与对照组相比,5-FU组HepG2/5-FU细胞EdU阳性率、A450值、划痕愈合率、P-gp、p-EGFR、p-FAK蛋白表达显著降低(P<0.05);与异钩藤碱组、5-FU组相比,异钩藤碱+5-FU组HepG2/5-FU细胞EdU阳性率、A450值、划痕愈合率、P-gp、p-EGFR、p-FAK蛋白表达降低(P<0.05);与异钩藤碱+5-FU组相比,异钩藤碱+5-FU+NSC228155组HepG2/5-FU细胞EdU阳性率、A450值、划痕愈合率、P-gp、p-EGFR、p-FAK蛋白显著上调(P<0.05)。结论异钩藤碱抑制HepG2细胞增殖、迁移及降低5-FU耐药性的机制可能与阻断EGFR/FAK通路有关。Obieetive To investigate the eliects of isorhynchophylline(isorhy)on the proliferation,migration,and 5-fU resistance of liver cancer cells by regulating the epidermal growth factor receptor(EGFR)/focal adhesion kinase(FAK)signaling pathway Methods Logarithmic phase HepG2 cells were divided into control group,low,medium,and high concentration groups of isorhy(32.5,65.0,130.0μmol·L^(-1)),and the high concentration group of isorhy(130μmol·L^(-1)l)+EGFR activator NSC228155(2μmol·L^(-1)).Logarithmic phase 5-FU-resistant hepatocellular carcinoma cell line HepG2/5-FU was divided into control group,isorhy(130μmol·L^(-1)l)group,5-FU(60μmol·L^(-1))group,isorhy(130μmol·L^(-1))+5-FU(60μmol·L^(-1))group,and isorhy(130μmol·L^(-1))+5-FU(60μmol-Ll)+NSC228155(22μmol·L^(-1)l)group.The cells were treated with drugs for 24 hours,and the control group was not treated.EdU staining and CCK-8 were applied to detect HepG2 or HepG2/5-FU cell proliferation.Scratch experiment was applied to detect the migration of HepG2 or HepG2/5-FU cells.Western blotting was applied to detect cyclinD1,MIEN1,P-glycoprotein(P-gp),p-EGFR,and p-FAK proteins in cells.Results Compared with the control group,the EdU positive rate,A4so value,scratch healing rate,CyclinD1,MIEN1,p-EGFR,and p-FAK protein expression of HepG2 cells in isorhy group were reduced,in a concentration-dependent manner(P<0.05).Compared with isorhy(130μmol·L^(-1))group,the EdU positive rate,A4so value,scratch healing rate,CyclinD1,MIEN1,p-EGFR,and p-FAK protein expression of HepG2 cells in isorhy(130.0μmol·L^(-1)l)+NSC228155 group were increased(P<0.05).Compared with control group,the EdU positive rate,scratch healing rate,A4so value,P-gp,p-EGFR,and p-FAK protein expression of HepG2/5-FU cells in the 5-FU group were reduced(P<0.05).Compared with the isorhy group or 5-FU group,the EdU positive rate,A4so value,scratch healing rate,P-gp,p-EGFR,and p-FAK protein expression of HepG2/5-FU cells in the isorhy+5-FU group were reduced(P<0.05).Compared with the isorhy+5-FU group,

关 键 词：异钩藤碱 肝癌 表皮生长因子受体/局部黏斑激酶(EGFR/FAK)通路 增殖 迁移 5-FU耐药性 P-糖蛋白(P-gp) 

分 类 号：R965[医药卫生—药理学]