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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。Western blotting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 study the effect of exosomes derived from lung cancer cells on the migration of secretory cells and their cognate tumor cells, and to explore the role of PI3K / Akt and SRC signal transduction pathways in this process. Methods Exosomes derived from lung cancer cells were isolated from lung cancer cell A549 supernatant by density gradient centrifugation. The morphology of Exosomes was observed by transmission electron microscopy and the protein expression was observed by Western blotting. Transwell assay detects cell migration ability. Results Transmission electron microscopy was used to observe the origin of lung cancer A549 cells Exosomes, which has a characteristic disk-like structure, composed of bilayer membrane diameter 30 ~ 100 nm. Western blotting showed that the surface of Exosomes was rich in CD9 molecules. A549-derived Exosomes promoted the migration of A549 and its cognate tumor cell HCC827 cells in a dose-dependent manner with Activated Akt and SRC. Conclusion Exosomes derived from lung cancer cells can promote the migration of their secretory cells and their cognate tumor cells. The mechanism may be related to the activation of Akt and SRC.