Elsevier

Life Sciences

Volume 265, 15 January 2021, 118757
Life Sciences

miR-202-3p overexpression attenuates endometriosis-like lesions by modulating YAP-dependent transcription of S100A6 in murine models

https://doi.org/10.1016/j.lfs.2020.118757Get rights and content

Abstract

Aim

Recent evidence has suggested the important implications of microRNAs (miRNAs) in the processes of proliferation and tissue remodeling in endometriosis (EMS). We therefore aim to determine the role of miR-202-3p in the pathophysiology of EMS and its underlying mechanisms.

Methods

Experimental endometriosis was induced in ovariectomized mice implanted with a slow-release 17-β estradiol capsule. Eutopic endometrial stromal cells (euESCs) were isolated and assayed for proliferative, invasive and apoptotic properties by EdU staining, Transwell assays, and flow cytometry. The invasive and apoptotic features in the endometrium of mice with EMS in vivo were evaluated by using immunohistochemical staining and TUNEL assays.

Results

miR-202-3p was observed to be downregulated in the endometrial tissues of EMS patients. MiR-202-3p was also found to target YAP1 which resulted in reduced euESC proliferation and invasion and increased apoptosis. YAP1 was able to phosphorylated STAT3 which consequently upregulated S100A6 to promote the proliferative and invasive abilities of euESCs. MiR-202-3p was thereby proposed to act as an inhibitor of proliferation and tissue damage in the in vivo setting of EMS, its effects however, were able to be counteracted byS100A6, which reversed the effects of miR-202-3p on tissue injury and cell proliferation.

Conclusion

Our data together evidenced that miR-202-3p targeted YAP1 to reduce STAT3-mediated S100A6 whereby preventing the progression of EMS.

Introduction

Endometriosis (EMS) is defined as an inflammatory disease that is characterized by the presence of endometrial tissues outside the uterine cavity. The disease is commonly associated with chronic pelvic pain and infertility [1]. It is estimated that approximately 5–10% of females across the world suffer from this disease during their reproductive years some studies of which show an incidence rate of 11% [2]. These values are usually underreported, as the definite diagnosis of endometriosis requires invasive surgical visualization. The growth and detachment of aberrantly located endometrium during the menstrual cycle leads to bleeding, inflammatory response, developing cysts and scar tissues as well as adhesions which may consequently cause intestinal problems, chronic pelvic inflammation and infertility [3]. Patients with EMS may are found to an increased risk in developing a variety of chronic diseases some of which include; ovarian and breast neoplasms, cutaneous malignant melanoma, asthma along with autoimmune, cardiovascular or atopic diseases [4]. Recent molecular genetic researches have revealed that circulating epithelial progenitor cells or stem cells contributing to potential regeneration of uterine endometrium may become trapped outside the uterine cavity post-menstruation. These trapped progenitor cells can form nascent glands by clonal expansion and recruit polyclonal stromal cells, which are associated with the body's immune surveillance and chronic inflammation in ectopic tissues [5]. Endometriotic stromal cells (ESCs) are the major cell type that has been found in endometriotic lesions. At a cytogenic level, ESCs have been found to contain several specific epigenetic alterations of key transcription factors, such as GATA-binding factor-6 [6]. Although there have been advances in the clinical management of EMS-induced pain and clinical pregnancy, current medication interventions such as danazol and depot progestogens may be associated with higher adverse effects while the surgical therapeutic approach has been found to result in a high recurrence rate [7,8]. Hence, we strongly believe that the identification of epigenetic alterations and molecular biomarkers in the pathophysiology of EMS is vital in order to produce more therapeutic interventions for this disease.

Non-coding RNAs (ncRNAs) are RNA molecules that are not translated into their protein products. Many types of ncRNAs exist which include microRNAs (miRNAs), long non-coding RNAs (lncRNAs) and short inhibitory RNAs, all of which play various roles in the body. Some ncRNAs have been found to contribute to pathological processes through regulation of inflammatory response, proliferation, angiogenic process and tissue remodeling [9]. MiR-202-3p is one example of miRNA that is overexpressed in severe preeclampsia [10], but has conversely been found to be under-expressed in endometrial tissues relative to healthy (control) endometrium [11]. A recent study has reported the close association between miR-202-3p expression and birth weight-for-gestational age (BWGA) z-score, suggesting a great value of biological information about fetal growth during pregnancy [12]. The Yes-associated protein 1 (YAP1) was identified as a key modulator of EMS [13], which also functions as a regulator to elevate STAT3 [14], which is aberrantly activated in the setting of EMS [15]. Meanwhile, genes such as CD63, S100A6, and GNB2L1 are found to be involved in the molecular pathways associated with EMS including apoptosis, angiogenesis, proliferation and maintenance of cellular homeostasis in the ectopic endometrium [16]. Furthermore, the abnormal activation of S100A6 is likely reported to be a contributor of EMS [17]. Based aforementioned facts, we speculate that those miRNA and mRNAs are likely to be play important roles and be responsible for the ectopic proliferation of cells during EMS. Since YAP1 is predicted to be a target of miR-202-3p, we therefore carry out this investigation to further understand whether miR-202-3p can affect the proliferation of ESCs through regulating YAP1/STAT3/S100A6.

Section snippets

Ethics statement

Written informed consent was obtained from all participants. The study was conducted with the approval of the Ethics Committee of Xiangya Hospital, Central South University. The animal experiments were conducted with the approval of the Animal Ethics Committee of Xiangya Hospital, Central South University.

Tissue collection

From August 2017 to November 2018, endometrial tissues were collected from 30 patients with EMS. Normal endometrial tissues were collected from 30 patients with tubal obstruction randomly

MiR-202-3p is under expressed in EMS

MiR-202-3p has been reported to be under-expressed in patients with EMS and its overexpression may prevent the occurrence of EMS [11]. In this investigation we consistently verified a lower miR-202-3p expression in endometrial tissues of patients with EMS compared to normal endometrial tissues by RT-qPCR (p < 0.05, Fig. 1A). Moreover, a mouse model of EMS was developed and identified by HE staining, which exhibited endometrial tissue necrosis, inflammatory cell infiltration, endometrial stroma

Discussion

There has been an increase in the number of researches that addresses the use of therapeutical use of miRNAs as ideal candidates that can serve as disease biomarkers for patients with EMS [26,27]. In one instance, the abnormal expression of miRNAs such as miR-139-5p and miR-375 in endometriotic lesions have been found to correlate with the pathogenesis and development of gynecological diseases [28]. In our study, we provided evidence for the anti-proliferative and pro-apoptotic effects of

Conclusion

In conclusion, miR-202-3p is addressed to be an important regulator of EMS by impairing EMS-induced proliferation. A regulatory network miR-202-3p/YAP1/STAT3/S100A6 proposed as a result of this should help in contributing to a better understanding of EMS pathogenic mechanism.

The following are the supplementary data related to this article.

. Bioinformatic analysis. A, 50 shared mRNAs among the TargetScan, TargetMiner, microT-CDS, miRDB databases, and the GSE7305 dataset. B, 2 shared

Funding

None.

Author contributions

Kangling Xi and Jing Lan designed the study. Kangling Xie and Jing Lan collated the data, carried out data analyses and produced the initial draft of the manuscript. Jing Lan contributed to drafting the manuscript. All authors have read and approved the final submitted manuscript.

Declaration of competing interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Acknowledgements

We would like to give our sincere appreciation to the reviewers for their helpful comments on this article.

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