摘要
目的:探究抗表皮生长因子受体二聚体界面的新抗体EGFR dimer 5G9对肿瘤细胞生长的抑制效果及机制。方法:选用表皮生长因子受体不同表达水平的3种细胞系进行研究;用MTT法检测EGFR dimer 5G9单抗对3种细胞体外生长的抑制效果;选取A431细胞作为表皮生长因子受体过表达模型,研究EGFR dimer 5G9单抗对表皮生长因子受体磷酸化的影响;用BALB/c小鼠荷人表皮鳞癌模型进行体内抑瘤实验。结果:表皮生长因子受体在NIH-3T3细胞低表达,而在BxPC-3细胞和A431细胞均过表达,EGFR dimer 5G9单抗对BxPC-3细胞和A431细胞的生长抑制率分别为(50.17±0.75)%和(47.09±0.94)%,而对NIH-3T3细胞没有明显的抑制作用;A431细胞在EGFR dimer 5G9单抗联合表皮生长因子的作用下,2 h后表皮生长因子受体磷酸化水平有所下调,并在12 h后磷酸化水平被显著抑制;体内抑瘤实验显示,EGFR dimer 5G9单抗给药组和对照组的裸鼠在实验结束时的平均肿瘤体积分别为(1726±187.6)和(3082.5±276.1)mm3,平均肿瘤湿重分别为(1.31±0.13)和(2.33±0.08)g,EGFR dimer 5G9单抗在体内有显著的抑瘤效果,抑瘤率达到43.78%。结论:靶向EGFR二聚体界面的新抗体EGFR dimer 5G9通过抑制表皮生长因子受体磷酸化来抑制表皮生长因子受体信号通路的活性,从而可有效抑制表皮生长因子受体过表达进而抑制肿瘤的生长。
Objective:To investigate the inhibitory effect and the mechanism of EGFR dimer 5G9,a new antibody against epidermal growth factor receptor(ECFR)dimer interface,on the growth of cancer cells.Methods:Three cell lines with different expression levels of EGFR were selected for this experiment.MTT assay was used to detect the inhibitory effect of the monoclonal antibody ECFR dimer 5G9 on the growth of the three cell lines in vitro.A431 cells were selected as the model of ECFR overexpression to study the effect of monoclonal antibody EGFR dimer 5G9 on EGFR phosphorylation.The tumor inhibition experiment in vivo was carried out in BALB/c mice bearing human epidermal squamous cell carcinoma.Results:ECFR was expressed in low level in NIH-3T3 cells,but overexpressed in BxPC-3 cells and A431 cells.The growth inhibitory rates of monoclonal antibody EGFR dimer 5G9 on BxPC-3 cells and A431 cells were(50.17±0.75)%and(47.09±0.94)%,respectively,but it had no significant inhibitory effect on NIH-3T3 cells.The phosphorylation levels of EGFR in A431 cells were down-regulated by monoclonal antibody ECFR dimer 5C9 combined with epidermal growth factor 2 h later,and was significantly inhibited 12 h later.In vivo anti-tumor test showed that the average tumor volume in the monoclonal antibody ECFR dimer 5G9 administration group and control group were(1726±187.6)mm²and(3082.5±276.1)mm,respectively.The average tumor wet weights were(1.31±0.13)and(2.33±0.08)g,respectively.Monoclonal antibody EGFR dimer 5G9 had significant tumor inhibition effect in vivo,and the tumor inhibition rate reached 43.78%.Conclusion:EGFR dimer 5G9,a novel antibody targeting EGFR dimer,can exert influence on EGFR signaling pathway by inhibiting the phosphorylation of EGFR,thus effectively inhibit the growth of tumor overexpressing EGFR.
作者
梁志文
赵林
李泽民
王爽
朱林峰
吴雪蔓
蔡静敏
郑可
李黄金
LIANG Zhi-wen;ZHAO Lin;LI Ze-min;WANG Shuang;ZHU Lin-feng;WU Xue-man;CAI Jing-min;ZHENG Ke;LI Huang-jin(School of Life Sciences and Biopharmaceutics,Guangdong Pharmaceutical University,Biopharmaceutical Technology and Quality Control Laboratory,Guangzhou 510006,China)
出处
《中国新药杂志》
CAS
CSCD
北大核心
2024年第3期285-291,共7页
Chinese Journal of New Drugs
基金
国家自然科学基金资助项目(81273423)
广东省自然科学基金资助项目(2015A030310120)
广东省高校重大科研资助项目(2016KZDXM042)。
关键词
表皮生长因子受体
二聚体界面
单克隆抗体
抑瘤作用
受体磷酸化
epidermal growth factor receptor
dimer interface
monoclonal antibody
anti-tumor effect
receptor phosphorylation
