上海交通大学学报（医学版）

上海交通大学学报（医学版） ›› 2017, Vol. 37 ›› Issue (6): 726-.doi: 10.3969/j.issn.1674-8115.2017.06.003

• 论著（基础研究） • 上一篇    下一篇

TWEAK调控巨噬细胞外泌体中miRNA-7的表达抑制卵巢癌 细胞侵袭和迁移的研究

李栋，胡媛，吴安玥，邱兴堤，邱丽华   

  1. 上海交通大学 医学院附属仁济医院妇产科，上海市妇科肿瘤重点实验室，上海 200127
  • 出版日期:2017-06-28 发布日期:2017-07-05
  • 通讯作者: 邱丽华，电子信箱：lilyqiulh@126.com
  • 作者简介:李栋（1992—），男，硕士生；电子信箱：donglee.og@hotmail.com
  • 基金资助:

    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）

Exosomal miRNA-7 from TWEAK-stimulated macrophages inhibiting the invasion and migration of ovarian cancer cells

LI Dong, HU Yuan, WU An-yue, QIU Xing-di, QIU Li-hua   

  1. Department of Gynecology and Obstetrics, Shanghai Key Laboratory of Gynecologic Oncology, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200127, China
  • Online:2017-06-28 Published:2017-07-05
  • Supported by:

    National Natural Science Foundation of China, 81272884; Foundation of Shanghai Municipal Commission of Health and Family Planning, ZHYYZXYJHZX-2-06; Shanghai Municipal Education Commission–Gaofeng Clinical Medicine Grant Support, 20161412

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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HO8910PM 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；AntagomiR-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。

关键词: &ensp, 上皮性卵巢癌, 巨噬细胞, 外泌体, TWEAK, 细胞侵袭和迁移

Abstract:

 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.

Key words: epithelial ovarian cancer, macrophages, exosome, TWEAK, cell invasion and migration