Department of Minimally Invasive Gynecologic Center, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, No. 17 Qi Helou Road, Dong Cheng District, Beijing, 100006, China.
Department of Urology, Beijing Tongren Hospital, Capital Medical University, Beijing, 100730, China.
Reprod Sci. 2021 May;28(5):1523-1539. doi: 10.1007/s43032-020-00444-8. Epub 2021 Feb 4.
Adenomyosis (ADS) is a commonly encountered benign gynecological disorder. Epithelial-mesenchymal transition (EMT) may serve a pivotal role in the pathogenesis of ADS. Talin1 has been identified to be implicated in multiple human carcinomas, probably through inducing EMT process. However, available data on the precise molecular mechanism of Talin1 in the pathogenesis of ADS remain extremely scanty. In the present study, we aim to investigate the clinical roles of Talin1 and its effects on uterine endometrial cell migration, invasion, and EMT in ADS. Relative mRNA expression of Talin1, microRNA-145-5p (miR-145-5p), and EMT-related markers was determined by qRT-PCR. Immunohistochemistry and immunofluorescence were performed to examine the distribution of Talin1 in ADS endometrium. Protein levels of Talin1, EMT-related markers, and wnt/β-catenin pathway were measured by western blot. Wound healing assay and transwell assay were utilized for evaluating cell migration and invasion respectively. Dual-luciferase reporter assay was performed to verify the relationship between Talin1 and miR-145-5p. We found Talin1 was markedly overexpressed in ADS endometrial tissue and cells, whereas miR-145-5p was downregulated. Elevated Talin1 mRNA level might be closely related to some clinicopathological features of ADS. Through functional experiments, we demonstrated that overexpression of Talin1 induced EMT and enhanced migration and invasion ability of ADS eutopic and ectopic endometrial epithelial cells (ADS_Eu_EEC and ADS_Ec_EEC) in vitro through activating the canonical wnt/β-catenin pathway. From a mechanistic perspective, Talin1 was inversely regulated by miR-145-5p as a direct target. Our findings unveiled that under the regulation of miR-145-5p, Talin1 might promote endometrial cell migration and invasion through inducing EMT, presenting a novel insight for elucidating the pathogenesis of ADS.
腺肌病（ADS）是一种常见的良性妇科疾病。上皮-间充质转化（EMT）可能在 ADS 的发病机制中起关键作用。Talin1 已被确定与多种人类癌有关，可能通过诱导 EMT 过程。然而，关于 Talin1 在 ADS 发病机制中的精确分子机制的可用数据仍然非常有限。在本研究中，我们旨在研究 Talin1 的临床作用及其对 ADS 中子宫子宫内膜细胞迁移、侵袭和 EMT 的影响。通过 qRT-PCR 测定 Talin1、微小 RNA-145-5p（miR-145-5p）和 EMT 相关标记物的相对 mRNA 表达。通过免疫组织化学和免疫荧光法检查 Talin1 在 ADS 子宫内膜中的分布。通过 Western blot 测定 Talin1、EMT 相关标记物和 wnt/β-catenin 通路的蛋白水平。利用划痕愈合试验和 Transwell 试验分别评估细胞迁移和侵袭。双荧光素酶报告基因实验验证 Talin1 与 miR-145-5p 之间的关系。我们发现 Talin1 在 ADS 子宫内膜组织和细胞中明显过表达，而 miR-145-5p 下调。升高的 Talin1 mRNA 水平可能与 ADS 的一些临床病理特征密切相关。通过功能实验，我们证明 Talin1 的过表达通过激活经典 wnt/β-catenin 通路，在体外诱导 ADS 在位和异位子宫内膜上皮细胞（ADS_Eu_EEC 和 ADS_Ec_EEC）的 EMT 并增强其迁移和侵袭能力。从机制上讲，Talin1 作为直接靶标受 miR-145-5p 的反向调节。我们的研究结果表明，在 miR-145-5p 的调节下，Talin1 可能通过诱导 EMT 促进子宫内膜细胞的迁移和侵袭，为阐明 ADS 的发病机制提供了新的见解。
