三阴性乳腺癌细胞膜表面EGFR和c-Met的单分子水平共定位研究  

Single-Molecule Colocalization of EGFR and c-Met on the Surface of Triple Negative Breast Cancer Cells

作　　者：柴彬彬 王丽[2] 李劲涛[1] 张晓菲 夏阳 吉元[2] 盛望[1] 韩晓东[2] CHAI Bin-Bin;WANG Li;LI Jin-Tao;ZHANG Xiao-Fei;XIA Yang;JI Yuan;SHENG Wang;HAN Xiao-Dong(College of Life Science and Bioengineering,Beijing University of Technology,Beijing 100124,China;Solid State Institute of Microstructure and Property of Advanced Materials,Beijing University of Technology,Beijing 100124,China)

机构地区：[1]北京工业大学生命科学与生物工程学院,北京100124 [2]北京工业大学固体微结构与性能研究所,北京100124

出　　处：《生物技术通讯》2020年第1期66-70,共5页Letters in Biotechnology

基　　金：北京结构生物学高精尖创新中心课题(997012045-100300001)。

摘　　要：目的:在纳米水平直观验证表皮生长因子受体(EGFR)和肝细胞生长因子受体(c-Met)是否可以结合形成异源二聚体。方法:采用链霉亲和素-生物素技术,利用2种不同大小的纳米金颗粒(10和30 nm)分别结合核酸适配体对EGFR和c-Met进行共标记和共定位;采用环境扫描电镜对2种膜蛋白进行纳米尺度下的超高分辨成像观察,并开展统计分析。结果:三阴性乳腺癌细胞膜表面EGFR单体、二聚体、多聚体的占比分别为48.60%、29.83%、21.57%,c-Met单体、二聚体、多聚体的占比分别为38.24%、27.66%、34.10%;EGFR、c-Met可以聚合形成异源二聚体和多聚体,分别占二聚体和多聚体总数的13.71%和24.42%。结论:EGFR和c-Met可以通过形成不同类型聚体的形式在膜上发挥作用,提示联合靶向EGFR和c-Met将成为针对三阴性乳腺癌的一种新的潜在治疗方法。Objective:To visually verify that epidermal growth factor receptor(EGFR)and cellular mesenchymalepithelial transition factor(c-Met)can combine to form heterodimers at the nanometer level.Methods:EGFR and c-Met were co-localized using two streptavidin-labelled gold nanoparticlesof different sizes(10 and 30 nm)respec tively combined with biotin-labelled EGFR and c-Met nucleic acid aptamers.Environmental scanning electron mi croscopy was used to observe the ultra-high resolution imaging of two membrane proteins at the nanoscale,and statistical analysis was performed.Results:The proportions of EGFR monomers,dimers,and multimers on the sur face of triple-negative breast cancer(TNBC)cell membranes were 48.60%,29.83%and 21.57%,respectively;the proportions of c-Met monomers,dimers,and multimers were 38.24%,27.66%and 34.10%,respectively.EGFR and c-Met can polymerize to form heterodimers and multimers,which account for 13.71%and 24.42%of the to tal number of dimers and multimers,respectively.Conclusion:EGFR and c-Met can play a role on the membrane by forming different types of aggregates,suggesting that the combined targeting of EGFR and c-Met will be come a new potential treatment method for TNBC.

关 键 词：三阴性乳腺癌 表皮生长因子受体(EGFR) 肝细胞生长因子受体(c-Met) 扫描电镜 共定位 

分 类 号：Q6-33[生物学—生物物理学] Q25