摘要
目的·检测TWEAK刺激后,巨噬细胞及其分泌的外泌体中miRNA-7的表达;探索TWEAK刺激后,巨噬细胞源性外泌体抑制上皮性卵巢癌细胞(HO8910-PM)侵袭和迁移的机制。方法·收集TWEAK刺激前后的巨噬细胞源性外泌体,并将其与HO8910-PM细胞共培养,real-time PCR检测巨噬细胞、巨噬细胞源性外泌体和共培养后HO8910-PM细胞中miRNA-7的表达;Western blotting检测共培养后HO8910-PM细胞中EGFR/AKT/ERK1/2信号通路的表达;Antagomi R-7处理降低巨噬细胞中miRNA-7表达后,收集TWEAK刺激前后的外泌体并进行transwell侵袭和迁移实验,观察HO8910-PM细胞侵袭和迁移能力的变化。结果·TWEAK刺激后,巨噬细胞内及其外泌体中miRNA-7的表达升高,并可上调HO8910-PM细胞中miRNA-7的表达,下调EGFR信号通路的表达。预先下调巨噬细胞中miRNA-7的表达后,巨噬细胞源性外泌体和共培养后HO8910-PM细胞中miRNA-7的表达降低,TWEAK刺激的巨噬细胞源性外泌体原先抑制HO8910-PM细胞转移的作用得到恢复。结论·miRNA-7在TWEAK刺激巨噬细胞分泌的外泌体中发挥重要作用,可通过抑制上皮性卵巢癌细胞中EGFR信号通路抑制其侵袭和迁移。
Objective · To determine the expression of miRNA-7 in TWEAK-stimulated macrophages and their secreted exosomes; to investigate the role of exosomal miRNA-7 from TWEAK-stimulated macrophages in modulating the metastasis of epithelial ovarian cancer （EOC） cells. Methods · Real-time PCR analysis was used to determine the miRNA-7 expression in TWEAK-stimulated macrophages, their exosomes and recipient HO8910-PM cells. The activity of EGFR signaling pathway in HO8910-PM cells was detected by Western blotting analysis. AntagomiR-7 was used to downregulate the miRNA-7 expressions in macrophage exosomes and then their effect on metastasis of HO8910-PM cells was examined by transwell assay. Results · TWEAK increased the levels of miRNA-7 in macrophages and their secreted exosomes and also resulted in an elevated level of miRNA-7 in recipient HO8910PM cells, which eventually reduced the activity of EGFR/AKT/ERK1/2 pathway. Pre-transfection of antagomiR-7 remarkably decreased the levels of miRNA-7 in macrophages, their secreted exosomes and the recipient EOC cells, with the enhancement of HO8910-PM metastasis. Conclusion · Exosomal miRNA-7 from TWEAK-stimulated macrophages plays a critical role in suppressing the metastasis of EOC cells by attenuation of EGFR signaling pathway.
出处
《上海交通大学学报（医学版）》
CSCD
北大核心
2017年第6期726-731,共6页
Journal of Shanghai Jiao tong University:Medical Science
基金
国家自然科学基金(81272884)
上海市卫生和计划生育委员会科研项目(ZHYY-ZXYJHZX-2-06)
上海市教育委员会高峰高原学科建设计划(20161412)~~
