Zhang Mengmeng, Wang Xi, Xia Xiaomeng, Fang Xiaoling, Zhang Tingting, Huang Fengying
Department of Obstetrics and Gynecology, The Second Xiangya Hospital, Central South University, Changsha, Hunan, 410000, P.R. China.
Department of Obstetrics and Gynecology, Women's Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, 310006, P.R. China.
Cell Death Discov. 2022 Apr 2;8(1):151. doi: 10.1038/s41420-022-00941-6.
Endometriosis (EMs) is a benign gynecological disorder showing some tumor-like migratory and invasive phenotypes. This study intended to investigate the role of microRNA-30c (miR-30c) in EMs, which is involved with B-cell lymphoma 9 (BCL9), an activator of the Wnt/β-catenin signaling pathway. EMs specimens were clinically collected for determination of miR-30c and BCL9 expression. Exosomes were isolated from endometrial epithelial cells (EECs), and the uptake of exosomes by ectopic EECs (ecto-EECs) was characterized using fluorescence staining and confocal microscopy. The binding of miR-30c to BCL9 was validated by dual-luciferase reporter assay. Artificial modulation (up- and down-regulation) of the miR-30c/BCL9/Wnt/CD44 regulatory cascade was performed to evaluate its effect on ecto-EEC invasion and migration, as detected by Transwell and wound healing assays. A mouse model of EMs was further established for in vivo substantiation. Reduced miR-30c expression and elevated BCL9 expression was revealed in EMs ectopic tissues and ecto-EECs. Normal EECs-derived exosomes delivered miR-30c to ecto-EECs to suppress their invasive and migratory potentials. Then, miR-30c was observed to inhibit biological behaviors of ecto-EECs by targeting BCL9, and the miR-30c-induced inhibitory effect was reversed by BCL9 overexpression. Further, miR-30c diminished the invasion and migration of ecto-EECs by blocking the BCL9/Wnt/CD44 axis. Moreover, miR-30c-loaded exosomes attenuated the metastasis of ecto-EEC ectopic nodules. miR-30c delivered by EECs-derived exosomes repressed BCL9 expression to block the Wnt/β-catenin signaling pathway, thus attenuating the tumor-like behaviors of ecto-EECs in EMs.
子宫内膜异位症（EMs）是一种良性妇科疾病，具有一些肿瘤样的迁移和侵袭表型。本研究旨在探讨微小RNA-30c（miR-30c）在EMs中的作用，其与Wnt/β-连环蛋白信号通路的激活剂B细胞淋巴瘤9（BCL9）有关。临床收集EMs标本以测定miR-30c和BCL9的表达。从子宫内膜上皮细胞（EECs）中分离出外泌体，并使用荧光染色和共聚焦显微镜对异位EECs（ecto-EECs）摄取外泌体的情况进行表征。通过双荧光素酶报告基因测定法验证miR-30c与BCL9的结合。对miR-30c/BCL9/Wnt/CD44调节级联进行人工调节（上调和下调），以评估其对ecto-EEC侵袭和迁移的影响，通过Transwell和伤口愈合试验进行检测。进一步建立了EMs小鼠模型进行体内验证。结果显示，EMs异位组织和ecto-EECs中miR-30c表达降低，BCL9表达升高。正常EECs来源的外泌体将miR-30c递送至ecto-EECs，以抑制其侵袭和迁移潜能。然后，观察到miR-30c通过靶向BCL9抑制ecto-EECs的生物学行为，而BCL9过表达可逆转miR-30c诱导的抑制作用。此外，miR-30c通过阻断BCL9/Wnt/CD44轴减少ecto-EECs的侵袭和迁移。此外，装载miR-30c的外泌体减弱了ecto-EEC异位结节的转移。EECs来源的外泌体递送的miR-30c抑制BCL9表达，阻断Wnt/β-连环蛋白信号通路，从而减弱EMs中ecto-EECs的肿瘤样行为。
