西妥昔单抗对人肝癌细胞HepG2的体外效应  

Effects of cetuximab against human hepatocellular carcinoma cells HepG2 in vitro

作　　者：叶伟亮[1] 林周[1] 郎小玲 乔春霞[1] 李新颖[1] 王立燕[1] 吕明[1] 黎燕[1] 

机构地区：[1]军事医学科学院基础医学研究所分子免疫学研究室,北京100850 [2]北京天广实生物技术股份有限公司,北京100176

出　　处：《军事医学》2011年第3期202-206,共5页Military Medical Sciences

基　　金：国家自然科学基金资助项目(30972807)

摘　　要：目的探讨西妥昔单抗(爱必妥,cetuximab,Erbitux)对人肝癌细胞HepG2的体外效应。方法流式细胞术检测HepG2细胞表面表皮生长因子受体(EGFR)的表达。细胞划痕及Transwell实验检测西妥昔单抗对HepG2细胞迁移能力的影响。Western印迹检测西妥昔单抗对HepG2细胞EGFR、AKT及ERK表达及活化的影响。碘化丙啶(propidium lodide,PI)染色检测西妥昔单抗对细胞周期的影响。结果流式细胞分析结果显示,HepG2细胞表面存在较高水平的EGFR表达;细胞划痕及Transwell结果表明,西妥昔单抗对HepG2细胞迁移能力具有明显的抑制效应;Western印迹结果证明西妥昔单抗能显著降低HepG2细胞内EGFR、AKT及ERK的磷酸化水平;细胞周期分析显示西妥昔单抗作用24 h剂量依赖性影响HepG2细胞周期进程,并将其阻抑在G0/G1期。结论西妥昔单抗可以抑制高表达EGFR的肝癌细胞系HepG2胞内关键信号蛋白的活化,阻滞其细胞周期,抑制肝癌细胞的迁移能力。Objective To investigate the effect of cetuximab against human hepatocellular carcinoma cells HepG2 in vitro.Methods The membrane expression of EGFR was detected by flow cytometry in HepG2 cells.Then the migration of HepG2 cells treated with cetuximab was examined by scratch wound assay and Transwell migration assay,respectively.The expression and activition of EGFR,AKT and ERK1/2 was detected by Western blotting assay,and cell cycle distribution stained with propidium iodide（PI）was analyzed also by flow cytometry.Results The human hepatic carcinoma cells HepG2 were screened for a high expression level of membrane EGFR using flow cytometry.Both the scratch wound and Transwell migration assays showed that cetuximab could significantly inhibit the migration of HepG2 cells.Meanwhile,cetuximab could down-regulate the expression level of p-EGFR（Tyr1068）,p-AKT（Ser473）and p-ERK1/2（Thr202/Tyr204）in HepG2 cells.Besides,incubating HepG2 with rising cetuximab concentrations for 24 h led to a dose-dependent arrest of the cells in the G0/G1-phase of the cell cycle.Conclusion Cetuximab could inhibit the activation of key signal proteins,depress the cell cycle and inhibit the migration of HepG2 cells with a high expession level of membrane EGFR.

关 键 词：受体 表皮生长因子 西妥昔单抗 肝肿瘤 细胞运动 细胞周期 

分 类 号：R735.7[医药卫生—肿瘤]