Department of Obstetrics, The Affiliated Hospital of Hangzhou Normal University, China.
Department of Obstetrics, The Affiliated Hospital of Hangzhou Normal University, China.
Tissue Cell. 2021 Apr;69:101474. doi: 10.1016/j.tice.2020.101474. Epub 2020 Dec 21.
MiR-124-3p regulates the biological function of endometrial cancer cells. However, the role of miR-124-3p in adenomyosis (AM) has not been reported. Hence, we hypothesized that miR-124-3p also regulated the development of AM. The expressions of miR-124-3p and Neuropilin 1 (NRP1) in AM endometrial tissues were evaluated by Quantitative Real-time-PCR (qPCR). Endometrial stromal cells (ESCs) were isolated from the eutopic endometrial tissue of women with AM and further identified using immunofluorescence. Then the target of miR-124-3p was predicted by Starbase V2.0 and verified by dual-luciferase assay. After transfection of miR-124-3p mimic, inhibitor, or NRP1 overexpression plasmids, the viability and migration of ESCs were measured by Cell counting kit -8 (CCK-8) and wound healing assays, respectively. The expressions of NRP1 and epithelial-stromal transformation (EST)-related factors were evaluated by Quantitative Real-time-PCR (qPCR) or Western blot. MiR-124-3p was down-regulated and NRP1 was up-regulated in AM eutopic endometrial tissues. NRP1 was targeted by miR-124-3p. The extracted ESCs were Vimentin-positive and Cytokeratin-negative. MiR-124-3p mimic decreased viability, migration, and the expressions of NRP1, Vimentin, N-cadherin, and matrix metalloproteinase (MMP-9) in ESCs while increasing the expression of E-cadherin. MiR-124-3p inhibitor and NRP1 overexpression had the contrary effect of miR-124-3p on ESCs. Furthermore, NRP1 overexpression offset the effect of miR-124-3p mimic on viability, migration, and expressions of NRP1 and EMT-related factors in ESCs. MiR-124-3p regulated the migration and EMT of ESCs by down-regulating NRP1.
miR-124-3p 调节子宫内膜癌细胞的生物学功能。然而,miR-124-3p 在子宫腺肌病(AM)中的作用尚未报道。因此,我们假设 miR-124-3p 也调节 AM 的发展。通过定量实时聚合酶链反应(qPCR)评估 AM 子宫内膜组织中 miR-124-3p 和 Neuropilin 1(NRP1)的表达。从 AM 患者的在位子宫内膜组织中分离子宫内膜基质细胞(ESCs),并通过免疫荧光进一步鉴定。然后使用 Starbase V2.0 预测 miR-124-3p 的靶标,并通过双荧光素酶报告基因实验验证。转染 miR-124-3p 模拟物、抑制剂或 NRP1 过表达质粒后,通过细胞计数试剂盒-8(CCK-8)和划痕愈合实验分别测量 ESCs 的活力和迁移。通过定量实时聚合酶链反应(qPCR)或 Western blot 评估 NRP1 和上皮间质转化(EST)相关因子的表达。AM 在位子宫内膜组织中 miR-124-3p 下调,NRP1 上调。NRP1 是 miR-124-3p 的靶标。提取的 ESCs 呈波形蛋白阳性和角蛋白阴性。miR-124-3p 模拟物降低 ESCs 的活力、迁移和 NRP1、波形蛋白、N-钙粘蛋白和基质金属蛋白酶(MMP-9)的表达,同时增加 E-钙粘蛋白的表达。miR-124-3p 抑制剂和 NRP1 过表达对 ESCs 的作用与 miR-124-3p 相反。此外,NRP1 过表达抵消了 miR-124-3p 模拟物对 ESCs 活力、迁移和 NRP1 及 EMT 相关因子表达的影响。miR-124-3p 通过下调 NRP1 调节 ESCs 的迁移和 EMT。