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- 2017
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
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Abstract:
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。
： 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
