外泌体 miR-22-3p 来源于腹腔巨噬细胞，通过调节 SIRT1/NF-κB 信号通路增强异位子宫内膜基质细胞的增殖、迁移和侵袭。

Exosomal miR-22-3p derived from peritoneal macrophages enhances proliferation, migration, and invasion of ectopic endometrial stromal cells through regulation of the SIRT1/NF-κB signaling pathway.

机构信息

Department of Obstetrics and Gynecology, Qilu Hospital of Shandong University, Jinan, China.

出版信息

Eur Rev Med Pharmacol Sci. 2020 Jan;24(2):571-580. doi: 10.26355/eurrev_202001_20033.

DOI:10.26355/eurrev_202001_20033

PMID:32016958

Abstract

OBJECTIVE

Exosomes play crucial roles in cell-cell communication, but few studies exist on the role of exosomal miRNA in the interaction between peritoneal macrophages (pMφ) and human ectopic endometrial stromal cells (eESCs) in endometriosis (EMS). This study aimed to identify which exosomal miRNAs are significantly differently produced from EMS pMφ and to investigate the functional role of exosomal miRNAs in eESCs.

PATIENTS AND METHODS

Exosomes were collected from the culture media of pMφ by differential centrifugation. Confocal microscopy was used to identify whether the exosomes secreted by pMφ can be delivered into eESCs. miRNA microarray and quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR) were used to identify which exosomal miRNAs were specifically elevated in pMφ-derived exosomes from EMS and delivered into eESCs via exosomes. The effect of pMφ-derived miR-22-3p on the biological function of eESCs was assessed by Cell Counting Kit-8 (CCK-8), wound-healing, and transwell chamber assays. Bioinformatics analysis and Luciferase reporter assay were used to detect the binding of exosomal miR-22-3p to the 3'untranslated region of SIRT1. Western blot was utilized to detect the activity of SIRT1/NF-κB pathway.

RESULTS

Exosomes secreted by pMφ can successfully be transported to eESCs. pMφ-derived exosomes from EMS promoted the proliferation, migration, and invasion of eESCs. MiR-22-3p was significantly increased in pMφ-derived exosomes from EMS and delivered from pMφ to eESCs via exosomes. Mechanistic analyses revealed that exosomal miR-22-3p from pMφ promoted the proliferation, migration, and invasion of eESCs by targeting SIRT1 and activating NF-κB pathway.

CONCLUSIONS

Exosomal miR-22-3p promotes the proliferation, migration, and invasion of eESCs by regulating SIRT1/NF-κB pathway and may serve as a novel target for the inhibition of EMS progression.

摘要

目的

外泌体在细胞间通讯中发挥着关键作用，但关于外泌体 miRNA 在子宫内膜异位症（EMS）中腹膜巨噬细胞（pMφ）与人类异位子宫内膜基质细胞（eESCs）相互作用中的作用的研究甚少。本研究旨在鉴定 EMS pMφ 中差异产生的外泌体 miRNA，并研究外泌体 miRNA 在 eESCs 中的功能作用。

方法

通过差速离心从 pMφ 的培养物中收集外泌体。使用共聚焦显微镜鉴定 pMφ 分泌的外泌体是否可以递送至 eESCs。使用 miRNA 微阵列和定量逆转录-聚合酶链反应（qRT-PCR）鉴定 EMS 来源的 pMφ 衍生的外泌体中特异性升高并通过外泌体递送至 eESCs 的外泌体 miRNA。通过细胞计数试剂盒-8（CCK-8）、划痕愈合和 Transwell 室测定评估 pMφ 衍生的 miR-22-3p 对 eESCs 生物学功能的影响。生物信息学分析和荧光素酶报告基因测定用于检测外泌体 miR-22-3p 与 SIRT1 的 3'非翻译区的结合。Western blot 用于检测 SIRT1/NF-κB 通路的活性。

结果

pMφ 分泌的外泌体可以成功递送至 eESCs。EMS 来源的 pMφ 衍生的外泌体促进了 eESCs 的增殖、迁移和侵袭。miR-22-3p 在 EMS 来源的 pMφ 衍生的外泌体中显著增加，并通过外泌体从 pMφ 递送至 eESCs。机制分析表明，pMφ 衍生的外泌体 miR-22-3p 通过靶向 SIRT1 并激活 NF-κB 通路促进 eESCs 的增殖、迁移和侵袭。