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mi R-7的表达;过表达Dicer,real-time PCR检测巨噬细胞中mi R-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mi R-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mi R-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mi R-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。
Objective·To investigate the mechanism of tumor necrosis factor like weak inducer of apoptosis（TWEAK） promoting macrophagederived exosomal miR-7 to epithelial ovarian cancer（EOC）.Methods·The expression of Dicer was detected by real-time PCR and Western blotting in TWEAK-stimulated macrophages.The expression of miR-7 was detected by real-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 expression of miR-7.Then TWEAK was used to stimulate the macrophage,and the expression of miR-7 in the macrophage and macrophage-derived exosome was detected by real-time PCR.The expression of key proteins in the nuclear factor-κB（NF-κB） pathway was detected by Western blotting in TWEAK-stimulated macrophage.After pretreatment of NF-κB inhibitor,Western blotting was used to detect the expression of Dicer and key proteins in the NF-κB pathway in TWEAKstimulated macrophage.Results·The expression of Dicer in macrophage was down-regulated after TWEAK stimulating.The expression of miR-7 was up-regulated in Dicer-silencing macrophage and macrophage-derived exosome.While the expression of miR-7 was down-regulated in the macrophage and the macrophage-derived exosome in Dicer-overexpressing macrophage after TWEAK stimulating.The expression 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 expression 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 expression of Dicer to promote macrophage-derived exosomal miR-7 to EOC cells.
作者
邱兴堤
吴安玥
李栋
洪祖蓓
顾李颖
邱丽华
QIU Xing-di;WU An-yue;LI Dong;HONG Zu-bei;GU li-ying;QIU Li-hua(Department of Obstetrics and Gynecology,Renji Hospital,Shanghai Jiao Tong University School of Medicin;Shanghai Key Laboratory of Gynecology Oncology,Shanghai 200127,China)
出处
《上海交通大学学报（医学版）》
CAS
CSCD
北大核心
2018年第7期740-744,共5页
Journal of Shanghai Jiao tong University:Medical Science
基金
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)~~
