外泌体对A549细胞增殖、凋亡及迁移功能的影响

发布时间：2018-03-10 13:37

本文选题：肺癌　切入点：外泌体　出处：《广东医学》2017年10期 　论文类型：期刊论文

【摘要】：目的观察肺癌细胞来源的外泌体对肿瘤细胞增殖、凋亡及迁移能力的影响。方法采用多步超速离心法从肺癌A549细胞的培养液上清中分离出肺癌细胞来源的外泌体,并通过透射电子显微镜观察外泌体形态,利用Western blot检测其表面跨膜蛋白的表达。采用MTT法、ELISA法分别检测外泌体对A549细胞增殖和凋亡的影响;采用Transwell法了解外泌体对A549细胞迁移能力的影响。结果观察所提取的外泌体大小均匀,形态规则,多为圆形及椭圆形膜性囊泡盘状结构,直径30~100 nm,其外周包被被膜,表面具有丰富的四次跨膜蛋白CD9、CD63等。MTT法提示外泌体对A549细胞具有促增殖作用,且其影响具有时间和剂量依赖特征(P0.05),ELISA法检测用外泌体共培育后的A549细胞的凋亡情况,结果显示外泌体在一定程度上抑制肿瘤细胞凋亡(F=365.496,P=0.000)。Transwell法观察显示,外泌体具有促进A549细胞迁移运动的作用(F=252.005,P=0.000)。结论肺癌细胞来源的外泌体能促进A549细胞增殖、减少其凋亡、增强其迁移运动的能力。
[Abstract]:Objective To observe the exosome sources of lung cancer cells on tumor cell proliferation, apoptosis and migration. Method using multi step ultracentrifugation separation of exosomes from lung cancer cells from the culture supernatant of lung cancer A549 cells, and the exosome morphology was observed by transmission electron microscope, to detect the expression of the surface membrane protein by Western blot. By MTT method, respectively, to detect the influence of exosomes on proliferation and apoptosis of A549 cells by ELISA; Transwell was used to understand the effect of exosomes on the migration of A549 cells. Results exosomes from the uniform size and regular shape, mostly round and oval membranous sac bubble disc structure, diameter 30~100 nm, peripheral coated film, four transmembrane protein CD9 has rich surface, CD63.MTT suggested that exosomes could promote proliferation of A549 cells, and the effect is dose and time Dependence (P0.05), apoptosis of ELISA was measured by exosomes were after incubation of A549 cells, results showed that exosomes inhibit the apoptosis of tumor cells to a certain extent (F=365.496, P=0.000).Transwell observation showed that exosomes can promote A549 cell migration function (F=252.005, P=0.000) conclusion. Lung cancer cells derived extracellular fitness promote A549 cell proliferation, reduce apoptosis, enhance the ability of migration.

【作者单位】：广州市红十字会医院心胸外科;暨南大学第一临床医学院;
【基金】：广州市产学研协同创新重大专项项目(编号:201604020063)
【分类号】：R734.2

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