Department of Gynecology, The Affiliated Hangzhou Hospital of Nanjing Medical University, Hangzhou, Zhejiang 310008, P.R. China.
Department of Reproductive Medicine, Shanghai Tenth People's Hospital, Affiliated to Tongji University, Shanghai 200003, P.R. China.
Int J Mol Med. 2020 Dec;46(6):2078-2088. doi: 10.3892/ijmm.2020.4749. Epub 2020 Oct 9.
The enhanced migratory ability of endometrial stromal cells (ESCs) is a key factor in the formation of functional endometrium‑like tissues outside the uterine cavity during endometriosis (EMS). Although accumulating evidence has suggested the importance of microRNAs (miRNAs) in the pathogenesis of EMS, the role of particular miRNAs in the invasiveness of ESCs remain poorly understood. In the present study, the function of miRNAs in the invasiveness of ESCs, along with the associated underlying mechanism involved, were investigated. Initially, the expression patterns of miRNAs in the ectopic and eutopic endometrium isolated from patients with EMS were analyzed using microarray. MicroRNA‑202‑5p (miR‑202) was selected for further study due to its previously reported suppressive effects on the invasion in various types of cancers. The expression of miR‑202 and K‑Ras in eutopic and ectopic endometrioma tissues were detected using reverse transcription‑quantitative PCR, immunohistochemistry and western blotting. The migration and invasion ability of ESCs was determined using wound healing and Transwell invasion assays, respectively. Compared with that from healthy individuals, miR‑202 expression was demonstrated to be lower in the eutopic endometrium from patients with EMS, which was even lower in ectopic endometrium. Functional experiments in primary ESCs revealed that enhanced miR‑202 expression suppressed the cell invasion and migration abilities, which was also accompanied with increased E‑cadherin and reduced N‑cadherin expression in ESCs, suggesting its potentially suppressive role in epithelial‑mesenchymal transition. K‑Ras is a well‑known regulator of the ERK signaling pathway that was shown to be directly targeted and negatively regulated by miR‑202. In addition, K‑Ras expression was found to be upregulated in the ectopic endometrium, where it correlated negatively with that of miR‑202. Knocking down K‑Ras expression mimicked the anti‑invasive effects of miR‑202 overexpression on ESCs, whilst K‑Ras overexpression attenuated the inhibitory role of miR‑202 overexpression in ESC invasion. The K‑Ras/Raf1/MEK/ERK signaling pathway was also blocked by miR‑202 overexpression. These findings suggested that miR‑202 inhibited ESC migration and invasion by inhibiting the K‑Ras/Raf1/MEK/ERK signaling pathway, rendering miR‑202 a candidate for being a therapeutic target for EMS.
子宫内膜基质细胞(ESCs)的迁移能力增强是子宫内膜异位症(EMS)患者宫腔外形成功能性子宫内膜样组织的关键因素。虽然越来越多的证据表明 microRNAs(miRNAs)在 EMS 的发病机制中具有重要作用,但特定 miRNAs 在 ESCs 侵袭性中的作用仍知之甚少。在本研究中,研究了 miRNAs 在 ESCs 侵袭性中的功能及其相关的潜在机制。首先,使用微阵列分析了从 EMS 患者异位和在位子宫内膜中分离的 miRNAs 的表达模式。由于其先前报道的对各种类型癌症侵袭的抑制作用,选择 microRNA-202-5p(miR-202)进行进一步研究。使用逆转录-定量 PCR、免疫组织化学和 Western blot 检测 miR-202 和 K-Ras 在在位和异位子宫内膜异位症组织中的表达。使用划痕愈合和 Transwell 侵袭测定分别确定 ESCs 的迁移和侵袭能力。与健康个体相比,发现 EMS 患者在位子宫内膜中 miR-202 的表达较低,异位子宫内膜中甚至更低。在原代 ESCs 中的功能实验表明,增强的 miR-202 表达抑制了细胞侵袭和迁移能力,这也伴随着 ESCs 中 E-钙粘蛋白的增加和 N-钙粘蛋白的减少,表明其在上皮-间充质转化中具有潜在的抑制作用。K-Ras 是 ERK 信号通路的一个众所周知的调节剂,被证明是 miR-202 的直接靶标和负调控因子。此外,在异位子宫内膜中发现 K-Ras 表达上调,与 miR-202 的表达呈负相关。敲低 K-Ras 表达模拟了 miR-202 过表达对 ESCs 的抗侵袭作用,而 K-Ras 过表达减弱了 miR-202 过表达对 ESC 侵袭的抑制作用。K-Ras/Raf1/MEK/ERK 信号通路也被 miR-202 过表达阻断。这些发现表明,miR-202 通过抑制 K-Ras/Raf1/MEK/ERK 信号通路抑制 ESC 迁移和侵袭,使 miR-202 成为 EMS 的治疗靶点。