乳腺癌mda-mb-231细胞源exosomes对血管内皮细胞egfr表达的 .docVIP

乳腺癌MDA-MB-231细胞源exosomes对血管内皮细胞EGFR表达的影响 谢莹珊，沈宜，隆霜，范维柯，姜蓉，陈黎 (重庆医科大学基础医学院病理生理教研室 干细胞与组织工程研究室，重庆 400016) 【摘要】 目的 研究人乳腺癌MDA-MB-231细胞源exosomes介导表皮生长因子受体（epidermal growth factor receptor，EGFR）对人脐静脉内皮细胞HUVECs EGFR表达影响，探讨肿瘤组织血管内皮细胞异常表达EGFR的机制。方法 超速离心及密度梯度离心法提取MDA-MB-231细胞源exosomes；免疫细胞化学法检测MDA-MB-231细胞EGFR的表达；Western blot法检测MDA-MB-231细胞、exosomes及HUVECs EGFR蛋白表达；免疫细胞化学法检测HUVECs与exosomes共培养24 h后EGFR的表达；RT-PCR法检测MDA-MB-231细胞与处理组HUVECs细胞 EGFR mRNA的表达。 结果 EGFR在MDA-MB-231细胞中呈高表达，Western blot结果显示MDA-MB-231细胞及exosomes可见分子量170kD的EGFR蛋白呈阳性反应带，HUVECs呈阴性反应带；HUVECs与exosomes共培养24 h后镜下可见实验组部分HUVECs细胞胞质有淡黄色或棕黄色颗粒，EGFR阳性表达率为21.4±3.1%，与对照组相比，差异具有显著性（P0.01）。RT-PCR结果显示，MDA-MB-231细胞EGFR mRNA表达阳性，而处理组HUVECs EGFR mRNA表达阴性。 结论 MDA-MB-231细胞源exosomes携带癌基因EGFR，并能介导其向周围血管内皮细胞转移，这可能是肿瘤内皮细胞异常表达EGFR的一种方式。 【关键词】MDA-MB-231；exosomes；血管内皮细胞；EGFR；影响 【中图分类号】R730.23 【文献标识码】A Effect of exosomes derived from Breast Cancer Cell Line MDA-MB-231 on EGFR expression in HUVECs XIE Yingshan,SHEN Yi,LONG Shuang,FAN Weike,JIANG Rong,CHEN Li Department of Pathophysiology,Institute of Stem Cell and Tissue Engineering,College of Basic Medicine,Chongqing Medical Univercity,Chongqing 400016,China 【Abstract】Objective:research the possible manner of abnormal expression EGFR of tumor endothelial cell. Methods: Exosomes were purified by serial ultracentrifugation and sugar density ultracentrifugation.The expression of EGFR in MDA-MB-231 were detected by immunocytochemistry and Western blotting;The expression of EGFR in exosomes and HUVECs were detected by Western blotting;The expression of EGFR in HUVECs with exosomes co-cultured 24 h were detected by immunocytochemistry;RT-PCR were used to detect the mRNA expression of EGFR in MDA-MB-231 and experimental group.Results:MDA-MB-231 cells expressed a releative level of EGFR.In the result of Western blotting,the positive reaction band was found in 170 kD which coincides with the molecular weight of EGFR in MDA-MB-231 and exosomes,and negative reacti