College of Clinical Medicine, Ningxia Medical University, Yinchuan, China.
Medical Experimental Center, General Hospital of Ningxia Medical University, Yinchuan, China.
Pathol Oncol Res. 2021 Apr 29;27:1609761. doi: 10.3389/pore.2021.1609761. eCollection 2021.
Intrauterine adhesion (IUA) is one of the most prevalent reproductive system diseases in females. MicroRNAs (miRNAs) are reported to be master regulators in a variety of diseases, including IUA, but the role of microRNA-543 (miR-543) in IUA remains to be elucidated. In this study, we observed that miR-543 was downregulated in transforming growth factor-beta (TGF-β)-treated endometrial stromal cells (ESCs). Functionally, we observed that miR-543 suppressed the migration, epithelial-to-mesenchymal transition (EMT), and inhibited expression of extracellular matrix (ECM) proteins in TGF-β-treated ESCs. Mechanistically, MAPK1 is targeted by miR-543 after prediction and screening. A luciferase reporter assay demonstrated that miR-543 complementarily binds with the 3' untranslated region of mitogen-activated protein kinase 1 (MAPK1), and western blot analysis indicated that miR-543 negatively regulates MAPK1 protein levels. In addition, results from rescue assays showed that miR-543 inhibits the migration and EMT of TGF-β-treated ESCs by targeting MAPK1. In addition, we observed that miR-543 inactivates the Wnt/β-catenin signaling pathway through inhibiting the phosphorylation of MAPK1 and β-catenin. Finally, we confirmed that miR-543 represses migration, EMT and inhibits levels of ECM proteins in TGF-β-treated ESCs by targeting the Wnt/β-catenin signaling pathway. Our results demonstrated that miR-543 suppresses migration and EMT of TGF-β-treated ESCs by targeting the MAPK and Wnt/β-catenin pathways.
宫腔粘连（IUA）是女性最常见的生殖系统疾病之一。据报道，微小 RNA（miRNAs）是多种疾病的主要调控因子，包括 IUA，但 miRNA-543（miR-543）在 IUA 中的作用仍有待阐明。在本研究中，我们观察到 miR-543 在转化生长因子-β（TGF-β）处理的子宫内膜基质细胞（ESCs）中下调。功能上，我们观察到 miR-543 抑制 TGF-β 处理的 ESCs 的迁移、上皮间质转化（EMT），并抑制细胞外基质（ECM）蛋白的表达。从预测和筛选中发现，MAPK1 是 miR-543 的靶标。荧光素酶报告基因分析表明，miR-543 与丝裂原活化蛋白激酶 1（MAPK1）的 3'非翻译区互补结合，Western blot 分析表明，miR-543 负调控 MAPK1 蛋白水平。此外，挽救实验结果表明，miR-543 通过靶向 MAPK1 抑制 TGF-β 处理的 ESCs 的迁移和 EMT。此外，我们观察到 miR-543 通过抑制 MAPK1 和 β-catenin 的磷酸化使 Wnt/β-catenin 信号通路失活。最后，我们通过靶向 Wnt/β-catenin 信号通路证实 miR-543 通过抑制 MAPK1 和 Wnt/β-catenin 信号通路抑制 TGF-β 处理的 ESCs 的迁移、EMT 和 ECM 蛋白水平。我们的研究结果表明，miR-543 通过靶向 MAPK 和 Wnt/β-catenin 通路抑制 TGF-β 处理的 ESCs 的迁移和 EMT。
