唾液腺腺样囊性癌来源的外泌体促进癌细胞的增殖、迁移及侵袭

Salivary adenoid cystic carcinoma-derived exosomes promote the proliferation,migration and invasion of cancer cells

目的:探索人唾液腺腺样囊性癌不同细胞系分泌的外泌体对其癌细胞增殖、迁移及侵袭能力的影响。方法:使用ExoEasy Maxi Kit (Qiagen,Hilden,Germany)试剂盒提取外泌体,通过透射电镜、纳米粒子跟踪分析(NTA)、蛋白免疫印迹Western blotting技术鉴定提取的肿瘤外泌体。并在外泌体的干预下,采用CCK-8法、平板克隆形成实验、细胞划痕及Transwell实验检测细胞增殖、迁移及侵袭能力。结果:透射电镜及纳米粒子跟踪分析(NTA)检测显示:提取的外泌体主要直径在30~150 nm之间,其形态呈典型的"茶托型"结构;Western blotting检测到外泌体特异性标志蛋白CD9、CD63及TSG101呈阳性表达。CCK-8法及平板克隆实验显示唾液腺腺样囊性癌来源的外泌体可增强其癌细胞增殖能力(P<0.05)。细胞划痕及Transwell实验表明其外泌体可增强癌细胞迁移及侵袭能力(P<0.05)。结论:唾液腺腺样囊性癌不同细胞系均分泌外泌体。唾液腺腺样囊性癌外泌体可增强其癌细胞增殖、迁移及侵袭能力。

Objective:To investigate the effects of exosomes secreted by different cell lines of human salivary gland adenoid cystic carcinoma on proliferation,migration and invasion.Methods:Exosomes were extracted by the ExoEasy Maxi Kit(Qiagen,Hilden,Germany) kit.The methods of transmission electron microscopy,nanoparticle tracking analysis(NTA) and Western blotting were used to identify the exosomes of salivary adenoid cystic carcinoma.Under the intervention of exosomes,CCK-8 method,plate colony formation assay,cell scratch and Transwell assay were used to detect ability of proliferation,migration and invasion.Results:Transmission electron microscopy and nanoparticle tracking analysis(NTA) showed that the diameter of extracted exosomes were mainly between 30~150 nm,and their morphology was the "tea tray type" typical structure.Western blotting detected exosome-specific marker proteins CD9,CD63 and TSG101 were positively expressed.The CCK-8 method and plate cloning experiments showed that salivary adenoid cystic carcinoma-derived exosomes enhanced the proliferation of cancer cells(P<0.05).Wound healing test and Transwell experiments showed that exosomes enhanced the migration and invasion of cancer cells(P<0.05).Conclusion:Two different cell lines of salivary adenoid cystic carcinoma secrete exosomes.And the exosomes of salivary adenoid cystic carcinoma can enhance the proliferation,migration and invasion of cancer cells.