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Down-regulated miR-124-3p enhanced the migration and epithelial-stromal transformation of endometrial stromal cells extracted from eutopic endometrium in subjects with adenomyosis by up-regulating Neuropilin 1
Tissue & Cell ( IF 2.7 ) Pub Date : 2020-12-21 , DOI: 10.1016/j.tice.2020.101474
Nan Huang 1 , Liyan Xu 1 , Yafen Qiu 1 , Jinlai Zhan 1 , Xiangjun Chen 1
Affiliation  

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.



中文翻译:

下调miR-124-3p通过上调Neuropilin 1增强子宫腺肌病患者在位子宫内膜中提取的子宫内膜基质细胞的迁移和上皮间质转化

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。

更新日期:2020-12-30
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