上海交通大学学报(医学版) ›› 2018, Vol. 38 ›› Issue (7): 740-.doi: 10.3969/j.issn.1674-8115.2018.07.004

• 论著·基础研究 • 上一篇    下一篇

TWEAK调控Dicer促使巨噬细胞源性外泌体分选miR-7至卵巢癌细胞的机制

邱兴堤,吴安玥,李栋,洪祖蓓,顾李颖,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:2018-07-28 发布日期:2018-07-30
  • 通讯作者: 邱丽华,电子信箱:lilyqiulh@126.com。
  • 作者简介:邱兴堤(1983—),男,硕士生;电子信箱:64033116@qq.com。
  • 基金资助:
    国家自然科学基金面上项目(80172884);上海市卫生和计划生育委员会公卫项目(15gwzk0701);上海市教育委员会高峰高原计划(20161412);浦东新区卫生和计划生育委员会卫生行业科研专项(PW2016E-2)

Mechanism of TWEAK promoting macrophage-derived exosomal miR-7 to epithelial ovarian cancer cell through regulating Dicer

QIU Xing-di, WU An-yue, LI Dong, HONG Zu-bei, GU li-ying, QIU Li-hua   

  1. Department of Obstetrics and Gynecology, Renji Hospital, Shanghai Jiao Tong University School of Medicine; Shanghai Key Laboratory of Gynecology Oncology, Shanghai 200127, China
  • Online:2018-07-28 Published:2018-07-30
  • Supported by:
    National Natural Science Foundation of China, 80172884; Foundation of Shanghai Municipal Commission of Health and Family Planning, 15gwzk0701; Shanghai Municipal Education Commission—Gaofeng Clinical Medicine Support, 20161412; Pudong New District Commission of Health and Family Planning Program, PW2016E-2

摘要: 目的·研究TWEAK通过何种途径促使巨噬细胞源性外泌体分选miR-7至卵巢癌细胞。方法·收集TWEAK刺激前后的巨噬细胞,real-time PCR和Western blotting检测巨噬细胞中Dicer的表达。在巨噬细胞中沉默Dicer,real-time PCR检测巨噬细胞及其外泌体中miR-7的表达;过表达Dicer, real-time PCR检测巨噬细胞中miR-7的表达。继而用TWEAK刺激巨噬细胞,real-time PCR检测巨噬细胞源性外泌体中miR-7的表达,Western blotting检测TWEAK刺激前后的巨噬细胞中核因子κB(nuclear factor-κB,NF-κB)信号通路的表达;继而将NF-κB信号通路抑制,Western blotting检测TWEAK刺激前后巨噬细胞中Dicer的表达。结果· TWEAK刺激后,巨噬细胞中Dicer的表达下调。沉默巨噬细胞中Dicer的表达,巨噬细胞及其外泌体中miR-7的表达均上调;而将巨噬细胞中的Dicer过表达,再用TWEAK刺激巨噬细胞后,与对照组相比,Dicer过表达组巨噬细胞及其外泌体中的miR-7表达降低。TWEAK刺激巨噬细胞后,NF-κB信号通路蛋白的表达上调;而抑制NF-κB信号通路,再用TWEAK刺激巨噬细胞后,Dicer的表达无明显改变。结论· TWEAK通过活化NF-κB通路,抑制Dicer表达,从而促进巨噬细胞分选miR-7至其外泌体。

关键词: 上皮性卵巢癌, 肿瘤坏死因子样凋亡微弱诱导剂, Dicer, 巨噬细胞, 外泌体

Abstract: Objective · To investigate the mechanism of tumor necrosis factor like weak inducer of apoptosis (TWEAK) promoting macrophage-derived exosomal miR-7 to epithelial ovarian cancer (EOC). Methods · The of Dicer was detectedreal-time PCR and Western blotting in TWEAK-stimulated macrophages. The of miR-7 was detectedreal-time PCR in the macrophage and macrophage-derived exosome after silencing Dicer in macrophage. While in Dicer-overexpressing macrophage, real-time PCR was used to detect the of miR-7. Then TWEAK was used to stimulate the macrophage, and the of miR-7 in the macrophage and macrophage-derived exosome was detectedreal-time PCR. The of key proteins in the nuclear factor-κB (NF-κB) pathway was detectedWestern blotting in TWEAK-stimulated macrophage. After pretreatment of NF-κB inhibitor, Western blotting was used to detect the of Dicer and key proteins in the NF-κB pathway in TWEAK-stimulated macrophage. Results · The of Dicer in macrophage was down-regulated after TWEAK stimulating. The of miR-7 was up-regulated in Dicer-silencing macrophage and macrophage-derived exosome. While the of miR-7 was down-regulated in the macrophage and the macrophage-derived exosome in Dicer-overexpressing macrophage after TWEAK stimulating. The of key proteins in the NF-κB pathway in macrophage was also up-regulated after TWEAK stimulating. After inhibition of NF-κB signaling pathway, the of Dicer was not significantly changed in TWEAK-stimulated macrophage compared to the control group. Conclusion · TWEAK can active NF-κB pathway and inhibit the of Dicer to promote macrophage-derived exosomal miR-7 to EOC cells.

Key words: epithelial ovarian cancer, tumor necrosis factor like weak inducer of apoptosis (TWEAK), Dicer, macrophage, exosome

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