Studies Using anin VitroModel Show Evidence of Involvement of Epithelial-Mesenchymal Transition of Human Endometrial Epithelial Cells in Human Embryo Implantation

Lipocalin 2 (LCN2) is an induced stressor that promotes the epithelial–mesenchymal transition (EMT). We previously demonstrated that the development of endometriosis in mice correlates with the secretion of LCN2 in the uterus. Here, we sought to clarify the relationship between LCN2 and EMT in endometrial epithelial cells and to determine whether LCN2 plays a role in endometriosis. Antibodies that functionally inhibit LCN2 slowed the growth of ectopic endometrial tissue in a mouse model of endometriosis, suggesting that LCN2 promotes the formation of endometriotic lesions. Using nutrient deprivation as a stressor, LCN2 expression was induced in cultured primary endometrial epithelial cells. As LCN2 levels increased, the cells transitioned from a round to a spindle-like morphology and dispersed. Immunochemical analyses revealed decreased levels of cytokeratin and increased levels of fibronectin in these endometrial cells, adhesive changes that correlate with induction of cell migration and invasion.Lcn2knockdown also indicated that LCN2 promotes EMT and migration of endometrial epithelial cells. Our results suggest that stressful cellular microenvironments cause uterine tissues to secrete LCN2 and that this results in EMT of endometrial epithelial cells, which may correlate with the development of ectopic endometriosis. These findings shed light on the role of LCN2 in the pathology of endometrial disorders.

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MFGE8 regulates TGF-β-induced epithelial mesenchymal transition in endometrial epithelial cells in vitro

Reproduction ◽

10.1530/rep-15-0585 ◽

2016 ◽

pp. 225-233 ◽

Cited By ~ 5

Author(s):

Liang Yu ◽

Rong Hu ◽

Claretta Sullivan ◽

R James Swanson ◽

Sergio Oehninger ◽

...

Keyword(s):

Epithelial Cells ◽

Milk Fat ◽

Epithelial Mesenchymal Transition ◽

Mesenchymal Transition ◽

Force Microscopy ◽

Atomic Force ◽

Milk Fat Globule ◽

Cell Changes ◽

Endometrial Epithelial Cells

This study investigated the role of milk fat globule-epidermal growth factor-factor 8 (MFGE8) in TGF-β-induced epithelial–mesenchymal transition (EMT) of endometrial epithelial cells. These were in vitro studies using human endometrial epithelial cells and mouse blastocysts. We investigated the ability of TGF-β to induce EMT in endometrial epithelial cells (HEC-1A) by assessment of cytological phenotype (by light and atomic force microscopy), changes in expression of the markers of cell adhesion/differentiation E- and N-cadherin, and of the transcription factor Snail (by immunofluorescence and immunoblotting), and competence to support embryo attachment in a mouse blastocyst outgrowth assay. We also studied the effects of E-cadherin expression in cells transfected by retroviral shRNA vectors specifically silencing MFGE8. Results demonstrated that TGF-β induced EMT as demonstrated by phenotypic cell changes, by a switch of cadherin expression as well as by upregulation of the expression of the mesenchymal markers Snail and Vimentin. Upon MFGE8 knockdown, these processes were interfered with, suggesting that MFGE8 and TGF-β together may participate in regulation of EMT. This study demonstrated for the first time that endometrial MFGE8 modulates TGF-β-induced EMT in human endometrium cells.

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Recepteur d'origine nantais contributes to the development of endometriosis via promoting epithelial‐mesenchymal transition of a endometrial epithelial cells

Journal of Cellular and Molecular Medicine ◽

10.1111/jcmm.16261 ◽

2021 ◽

Author(s):

Qin Yu ◽

Jianzhang Wang ◽

Tiantian Li ◽

Xinyue Guo ◽

Shaojie Ding ◽

...

Keyword(s):

Epithelial Cells ◽

Epithelial Mesenchymal Transition ◽

Mesenchymal Transition ◽

Endometrial Epithelial Cells

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Versican V1 in human endometrial epithelial cells promotes BeWo spheroid adhesion in vitro

Reproduction ◽

10.1530/rep-18-0333 ◽

2019 ◽

pp. 53-64 ◽

Cited By ~ 4

Author(s):

Yumiko Miyazaki ◽

Akihito Horie ◽

Hirohiko Tani ◽

Masashi Ueda ◽

Asuka Okunomiya ◽

...

Keyword(s):

Epithelial Cells ◽

Chondroitin Sulfate ◽

Cell Line ◽

Human Embryo ◽

Embryo Implantation ◽

Cancer Cell Line ◽

Promoting Effect ◽

Ishikawa Cells ◽

Endometrial Epithelial Cells

The endometrium extracellular matrix (ECM) is essential for embryo implantation. Versican, a large chondroitin sulfate proteoglycan that binds hyaluronan and forms large ECM aggregates, can influence fundamental physiological phenomena, such as cell proliferation, adhesion and migration. The present study investigated the possible role of versican in human embryo implantation. Versican V1 expression and secretion in human endometrial epithelial cells (EECs) was most prominent in the mid-secretory phase. Versican expression in EECs significantly increased after treatment with estrogen and progesterone, but not by estrogen alone. We also established versican V1-overexpressing Ishikawa (endometrial cancer cell line) cells (ISKW-V1), versican V3-overexpressing (ISKW-V3) and control GFP-overexpressing (ISKW-GFP) Ishikawa cells. By the in vitro implantation model, the attachment ratio of BeWo (choriocarcinoma cell line) spheroids to the monolayer of ISKW-V1, but not of ISKW-V3, was found significantly enhanced compared with attachment to the ISKW-GFP monolayer. The conditioned medium derived from ISKW-V1 (V1-CM) also promoted the attachment of BeWo spheroids to the ISKW monolayer. However, this attachment-promoting effect was abolished when V1-CM was pretreated with chondroitinase ABC, which degrades chondroitin sulfate. Therefore, out of the ECM components, versican V1 may facilitate human embryo implantation.

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TET1 may contribute to hypoxia-induced epithelial to mesenchymal transition of endometrial epithelial cells in endometriosis

PeerJ ◽

10.7717/peerj.9950 ◽

2020 ◽

Vol 8 ◽

pp. e9950

Author(s):

Jingni Wu ◽

Xidie Li ◽

Hongyan Huang ◽

Xiaomeng Xia ◽

Mengmeng Zhang ◽

...

Keyword(s):

Epithelial Cells ◽

Epithelial To Mesenchymal Transition ◽

Epithelial Mesenchymal Transition ◽

Epigenetic Modification ◽

Hypoxia Inducible Factor ◽

Hypoxic Condition ◽

Mesenchymal Transition ◽

Global Increase ◽

Endometrial Epithelial Cells ◽

E Cadherin

Background Endometriosis (EMs) is a non-malignant gynecological disease, whose pathogenesis remains to be clarified. Recent studies have found that hypoxia induces epithelial-mesenchymal transition (EMT) as well as epigenetic modification in EMs. However, the relationship between EMT and demethylation modification under hypoxia status in EMs remains unknown. Methods The expression of N-cadherin, E-cadherin and TET1 in normal endometria, eutopic endometria and ovarian endometriomas was assessed by immunohistochemistry and immunofluorescence double staining. 5-hmC was detected by fluorescence-based ELISA kit using a specific 5-hmC antibody. Overexpression and inhibition of TET1 or hypoxia-inducible factor 2α (HIF-2α) were performed by plasmid and siRNA transfection. The expression of HIF-2α, TET1 and EMT markers in Ishikawa (ISK) cells (widely used as endometrial epithelial cells) was evaluated by western blotting. The interaction of HIF-2α and TET1 was analyzed by chromatin immunoprecipitation. Results Demethylation enzyme TET1 (ten-eleven translocation1) was elevated in glandular epithelium of ovarian endometrioma, along with the activation of EMT (increased expression of N-cadherin, and decreased expression of E-cadherin) and global increase of epigenetic modification marker 5-hmC(5-hydroxymethylcytosine). Besides, endometriosis lesions had more TET1 and N-cadherin co-localized cells. Further study showed that ISK cells exhibited enhanced EMT, and increased expression of TET1 and HIF-2α under hypoxic condition. Hypoxia-induced EMT was partly regulated by TET1 and HIF-2α. HIF-2α inhibition mitigated TET1 expression changes provoked by hypoxia. Conclusions Hypoxia induces the expression of TET1 regulated by HIF-2α, thus may promote EMT in endometriosis.

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