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目的探索肺癌细胞分泌的Exosomes对其分泌细胞及其同源肿瘤细胞增殖的影响,以及PI3K/Akt和SRC信号通路在其中的作用。方法采用密度梯度离心法从肺癌A549细胞的上清液中提取Exosomes,透射电子显微镜法观察Exosomes的形态,Western blotting法检测Exosomes标志蛋白的表达,MTT法检测细胞的增殖能力。结果 A549细胞来源的Exosomes的直径在30~100 nm之间,由双层膜构成。Western blotting检测到Exosomes中CD9的表达,且A549细胞来源的Exosomes以剂量和时间依赖性的方式促进其自身及同源肿瘤细胞HCC827细胞的增殖,并伴随Akt和SRC的活化。结论肺癌A549细胞来源的Exosomes以时间和剂量依赖性促进其自身及同源肿瘤细胞的增殖,其机制可能与Akt和SRC的活化有关。
Objective To explore the effects of exosomes secreted by lung cancer cells on the proliferation of their secretory cells and their cognate tumor cells and the role of PI3K / Akt and SRC signaling pathways. Methods Exosomes were extracted from the supernatant of lung cancer A549 cells by density gradient centrifugation. The morphology of Exosomes was observed by transmission electron microscopy. The expression of Exosomes was detected by Western blotting. The proliferation of A549 cells was detected by MTT assay. Results The diameter of A549-derived Exosomes was between 30 and 100 nm and consisted of bilayer membranes. CD9 expression in Exosomes was detected by Western blotting, and A549 cell-derived Exosomes promoted the proliferation of HCC827 cells in their own and homologous tumor cells in a dose- and time-dependent manner with the activation of Akt and SRC. Conclusion Exosomes derived from lung cancer A549 cells promote the proliferation of their own and homologous tumor cells in a time and dose dependent manner, which may be related to the activation of Akt and SRC.