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，尚未报道 miR-124-3p 在子宫腺肌病 (AM) 中的作用。因此，我们假设 miR-124-3p 也调节 AM 的发展。

通过定量实时 PCR (qPCR) 评估 miR-124-3p 和 Neuropilin 1 (NRP1) 在 AM 子宫内膜组织中的表达。从患有 AM 的女性的在位子宫内膜组织中分离出子宫内膜基质细胞 (ESC)，并使用免疫荧光进一步鉴定。然后通过Starbase V2.0预测miR-124-3p的靶标并通过双荧光素酶测定验证。转染 miR-124-3p 模拟物、抑制剂或 NRP1 过表达质粒后，分别通过细胞计数试剂盒 -8 (CCK-8) 和伤口愈合试验测量 ESC 的活力和迁移。通过定量实时 PCR (qPCR) 或蛋白质印迹评估 NRP1 和上皮-基质转化 (EST) 相关因子的表达。

在 AM 在位子宫内膜组织中，MiR-124-3p 下调，NRP1 上调。NRP1 被 miR-124-3p 靶向。提取的 ESC 为波形蛋白阳性和细胞角蛋白阴性。MiR-124-3p 模拟物降低了胚胎干细胞中 NRP1、波形蛋白、N-钙粘蛋白和基质金属蛋白酶 (MMP-9) 的活力、迁移和表达，同时增加了 E-钙粘蛋白的表达。MiR-124-3p 抑制剂和 NRP1 过表达对 ESC 具有 miR-124-3p 的相反作用。此外，NRP1 过表达抵消了 miR-124-3p 模拟物对 ESC 中 NRP1 和 EMT 相关因子的活力、迁移和表达的影响。

MiR-124−3p 通过下调 NRP1 来调节 ESC 的迁移和 EMT。