Akt和SRC在Exosomes促进同源肺癌细胞迁移中的作用

摘要

目的 研究肺癌细胞来源的Exosomes对其分泌细胞及同源肿瘤细胞迁移的影响,初步探讨PI3K/Akt和SRC信号转导通路在此过程中的作用.方法 采用密度梯度离心法,从肺癌细胞A549的上清液中分离出肺癌细胞来源的Exosomes,利用透射电子显微镜观察Exosomes的形态,用Western blotting实验观察蛋白的表达.Transwell实验检测细胞的迁移能力.结果 透射电子显微镜下观察肺癌A549细胞来源的Exosomes,其具有特征性的盘状结构,由双层膜构成,直径30～100 nm.Westernblotting结果显示Exosomes表面富含CD9分子.A549细胞来源的Exosomes以剂量依赖性的方式促进A549和其同源肿瘤细胞HCC827细胞的迁移,并伴随Akt和SRC的活化.结论 肺癌细胞来源的Exosomes能促进其分泌细胞和同源肿瘤细胞的迁移,其机制可能与Akt和SRC的活化有关.%Objective To investigate the effect of Exosomes derived from lung cancer cells on the migration of secretory cells and homologous tumor cells and to explore the role of PI3K/Akt and SRC signaling pathways in this process.Methods Exosomes were isolated from the supematant post density gradient centrifugation of A549,lung cancer cells.Morphology of the Exosomes was studied using transmission electron microscopy.Protein expression was analyzed using Western blotting.Cell migration was analyzed by a transwell assay.Results The double-membrane-bound Exosomes appeared as discal-shaped structures,30-100 nm in diameter.Western blotting showed that CD9 was abundant in the Exosomes.The Exosomes promoted the migration of A549 cells and their homologous tumor cells,HCC827 in a dose-dependent manner,accompanied by the activation of Akt and SRC.Conclusion The Exosomes derived from A549 (lung cancer) cells promote the migration of the secreting cells and the homologous tumor cells.The mechanism may be correlated with the activation of Akt and SRC.