阿法替尼调控Egr-1/TLR4/mTOR信号通路抑制肝癌细胞Hep3B增殖迁移侵袭及促进细胞凋亡的机制  

Study on the mechanism of afatinib regulating Egr-1/TLR4/mTOR signaling pathway to inhibit the proliferation,migration and invasion of hepatoma cells Hep3B and promote apoptosis

作　　者：李晶[1] 赵静[1] 孟德峰 LI Jing;ZHAO Jing;MENG De-feng(Department of hepatobiliary surgery,Kailuan general hospital,Tangshan Hebei,063000,China;不详)

机构地区：[1]开滦总医院肝胆外科,河北唐山063000 [2]华北理工大学附属医院

出　　处：《中西医结合肝病杂志》2021年第10期917-921,共5页Chinese Journal of Integrated Traditional and Western Medicine on Liver Diseases

摘　　要：目的:探讨阿法替尼是否通过早期生长反应因子1(Egr1)/Toll样受体4(TLR4)/哺乳动物雷帕霉素靶蛋白(mTOR)信号通路,抑制肝癌细胞Hep3B增殖迁移侵袭并促进其细胞凋亡。方法:采用不同浓度的阿法替尼处理Hep3B细胞,用免疫印迹试验(Western Blot)检测细胞周期蛋白D1(CyclinD1)、活化的(Cleaved)-天冬氨酸特异性半胱氨酸蛋白酶-3(caspase-3)、基质金属蛋白酶(MMP)-2、MMP9、Egr-1、TLR4、mTOR蛋白的表达,用噻唑蓝(MTT)法检测细胞增殖,流式细胞仪分析细胞凋亡,Transwell测定细胞迁移侵袭。在Hep3B细胞中转染pcDNA-Egr-1,并使用2.0μmol/L阿法替尼处理,采用上述方法检测细胞的增殖、迁移侵袭和凋亡。结果:阿法替尼显著降低Hep3B细胞的CyclinD1、MMP2、MMP9、Egr-1、TLR4、mTOR蛋白含量、增殖活性、细胞迁移及侵袭数量,显著提高Cleaved-caspase-3蛋白含量和凋亡率(P<0.05)。pcDNA-Egr-1可以逆转阿法替尼对Hep3B细胞增殖、迁移侵袭、凋亡和Egr-1/TLR4/mTOR信号通路的影响。结论:阿法替尼可以抑制肝癌细胞Hep3B的增殖,迁移侵袭,并诱导其凋亡,其机制与下调Egr-1/TLR4/mTOR信号通路有关。Objective:To investigate whether afatinib can inhibit the proliferation of hepatocellular carcinoma Hep3B through Egr-1/TLR4/mTOR signaling pathway,migrate and invade and promote apoptosis.Methods:Hep3B cells were treated with different concentrations of afatinib,and Western Blot was used to detect cyclin D1(CyclinD1)and activated(Cleaved)-aspartic acid-specific caspase-3(caspase)-3,matrix metalloproteinase(MMP)2,MMP9,Egr-1,TLR4,mTOR protein expression,thiazole blue(MTT)method to detect cell proliferation,flow cytometry analysis of cell apoptosis,Transwell measurement of cell migration and invasion.PcDNA-Egr-1 was transfected in Hep3B cells and treated with 2.0μmol/L afatinib.The above methods were used to detect cell proliferation,migration,invasion and apoptosis.Results:Afatinib significantly reduced CyclinD1,MMP2,MMP9,Egr-1,TLR4,mTOR protein content,proliferation activity,cell migration and invasion of Hep3B cells,and significantly increased the content of Cleaved-caspase-3 protein and the rate of apoptosis(P<0.05).pcDNA-Egr-1 can reverse the effects of afatinib on proliferation,migration,invasion,apoptosis and Egr-1/TLR4/mTOR signaling pathway of Hep3B cells.Conclusion:Afatinib can inhibit the proliferation,migration and invasion of hepatoma cells Hep3B,and induce its apoptosis.The mechanism is related to the down-regulation of Egr-1/TLR4/mTOR signaling pathway.

关 键 词：阿法替尼 Egr-1/TLR4/mTOR信号通路 肝癌细胞 增殖 迁移侵袭 凋亡 

分 类 号：R735.7[医药卫生—肿瘤]