摘要
目的探讨食管癌KYSE410细胞来源的外泌体对肿瘤细胞迁移及侵袭能力的影响及其机制。方法超速离心法分离食管癌KYSE410细胞培养上清中的外泌体;采用透射电镜及Western blotting观察所获得的外泌体的形态及其标志物;共聚焦显微镜观察KYSE410、KYSE510和YES2细胞摄取荧光染料标记的KYSE410外泌体;采用Transwell小室实验分析3种食管癌细胞的迁移及侵袭能力以及KYSE410外泌体对3种食管癌细胞迁移及侵袭能力的影响;Western blotting检测Wnt/β-catenin和PI3K/Akt信号通路相关蛋白的变化。结果透射电镜下观察KYSE410外泌体具有膜结构,直径分布在30~100nm;3种食管癌细胞的迁移及侵袭能力从大到小依次为KYSE410、KYSE510、YES2;KYSE410外泌体能够促进KYSE410、KYSE510、YES2细胞迁移及侵袭能力并增强3种食管癌细胞中β-catenin和p-Akt的表达。结论来源于高迁移及侵袭能力的食管癌KYSE410细胞的外泌体能够促进自身和低迁移及侵袭能力的食管癌KYSE510、YES2细胞的迁移和侵袭,并且可能通过激活Wnt/β-catenin和PI3K/Akt信号通路发挥上述作用。
Objective To investigate the biological effects ofexosomes secreted by KYSE410 cells on migration and invasion of KYSE410, KYSE510, YES2 cells and the possible mechanisms underlying the phenotype change. Methods The exosomes were isolated from the conditional supernatant of esophageal cancer cell line KYSE410 by ultracentrifugation. The morphology of exosomes was observed by transmission electron microscopy （TEM）. Western blotting was used to detect the protein markers of exosomes. The uptaken of fluorescence-labeled KYSE410 exosomes by KYSE410, KYSE510 and YES2 was also recorded under confocal microscopy. Migration and invasion ability of the three esophageal carcinoma cell lines and the effects of exosomes from KYSE410 on migration and invasion of KYSE410, KYSES10 and YES2 cells were analyzed by Transwell chamber, respectively. The alteration of Wnt/β-catenin and PI3K/Akt pathway-related proteins were detected by Western blotting. Results The membrane structure of KYSE410 derived exosomes could be observed with its diameter ranged between 30-100nm. The invasion and migration ability of three esophageal cancer cells are KYSE410〉 KYSE510〉 YES2. KYSE410 exosomes promoted the migration and invasion of KYSE410, KYSES10 and YES2 cells. Conclusions Concentrated exosomes derived from the highly migratory and invasive esophageal cancer cell line KYSE410 promoted the migration and invasion potentials of itself and esophageal cancer cell lines KYSE510 and YES2, which possibly exerted the effects by activating Wnt/β-catenin and PI3K/Akt signaling pathways.
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2017年第4期307-313,共7页
Medical Journal of Chinese People's Liberation Army
基金
国家自然科学基金(81572842
81672459)
中国医学科学院医学与健康科技创新工程项目(2016-12M-1-001)~~
关键词
食管癌
外泌体
肿瘤迁移
肿瘤侵袭
esophageal cancer
exosome
cancer migration
cancer invasion
