Hypo-Expression of Tuberin Promotes Adenomyosis via the mTOR1-Autophagy Axis

Adenomyosis (AM) is a disease in which endometrial tissue invades the myometrium and has a 10–60% prevalence in reproductive-aged women. TSC2 regulates autophagy via mTOR1 signalling in colorectal cancer and endometrial carcinoma. Dysregulation of autophagy is implicated in adenomyosis pathogenesis. However, whether TSC2 participates in adenomyosis via autophagy remains obscure. Here, we found that the expression of TSC2 in adenomyosis was significantly decreased than that in normal endometrium during the secretory phase. Moreover, TSC2 and autophagy marker expression was significantly lower in ectopic lesions than in eutopic samples. TSC2 downregulation inhibited autophagy through mTOR1 signalling pathway activation in endometrial cells, leading to excessive proliferation, migration, and EMT; TSC2 overexpression induced the opposite effects. Rapamycin treatment suppressed cell proliferation, migration and EMT in the absence of TSC2. In parallel, an autophagy-specific inhibitor (SAR-405) restored migration and EMT under rapamycin treatment in TSC2-knockdown Ishikawa cells. Finally, SAR-405 treatment promoted EMT and migration of overexpressing cells. Collectively, our results suggest that TSC2 controls endometrial epithelial cell migration and EMT by regulating mTOR1-autophagy axis activation and that hypo-expression of TSC2 in the endometrium might promote adenomyosis.

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Mouse double minute homologue 2 (MDM2) downregulation by miR-661 impairs human endometrial epithelial cell adhesive capacity

Reproduction Fertility and Development ◽

10.1071/rd17095 ◽

2018 ◽

Vol 30 (3) ◽

pp. 477 ◽

Cited By ~ 7

Author(s):

Amy Winship ◽

Amanda Ton ◽

Michelle Van Sinderen ◽

Ellen Menkhorst ◽

Katarzyna Rainczuk ◽

...

Keyword(s):

Gene Expression ◽

Cell Adhesion ◽

Epithelial Cell ◽

Epithelial Cells ◽

Endometrial Cells ◽

Endometrial Epithelial Cell ◽

Mouse Double Minute ◽

Double Minute ◽

Ishikawa Cells ◽

Endometrial Epithelial Cells

Human blastocysts that fail to implant following IVF secrete elevated levels of miR-661, which is taken up by primary human endometrial epithelial cells (HEECs) and impairs their adhesive capability. MicroRNA miR-661 downregulates mouse double minute homologue 2 (MDM2) and MDM4 in other epithelial cell types to activate p53; however, this has not been examined in the endometrium. In this study MDM2 protein was detected in the luminal epithelium of the endometrium, the site of blastocyst attachment, during the mid secretory receptive phase of the menstrual cycle. The effects of miR-661 on gene expression in and adhesion of endometrial cells was also examined. MiR-661 overexpression consistently downregulated MDM2 but not MDM4 or p53 gene expression in the Ishikawa endometrial epithelial cell line and primary HEEC. Adhesion assays were performed on the real-time monitoring xCELLigence system and by co-culture using Ishikawa cells and HEECs with HTR8/SVneo trophoblast spheroids. Targeted siRNA-mediated knockdown of MDM2 in endometrial epithelial cells reduced Ishikawa cell adhesion (P < 0.001) and also reduced HTR8/SVneo trophoblast spheroid adhesion to Ishikawa cells (P < 0.05) and HEECs (P < 0.05). MDM2 overexpression using recombinant protein treatment resulted in enhanced HTR8/SVneo trophoblast spheroid adhesion to Ishikawa cells (P < 0.01) and HEECs (P < 0.05). This study highlights a potential new mechanism by which human blastocyst-secreted miR-661 reduces endometrial epithelial cell adhesion; via downregulation of MDM2. These findings suggest that MDM2 contributes to endometrial–blastocyst adhesion, implantation and infertility in women.

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Different macrophages equally induce EMT in endometria of adenomyosis and normal

Reproduction ◽

10.1530/rep-17-0174 ◽

2017 ◽

Vol 154 (1) ◽

pp. 79-92 ◽

Cited By ~ 7

Author(s):

Min An ◽

Dong Li ◽

Ming Yuan ◽

Qiuju Li ◽

Lu Zhang ◽

...

Keyword(s):

Epithelial Cells ◽

Transforming Growth Factor ◽

Epithelial Mesenchymal Transition ◽

Quantitative Polymerase Chain Reaction ◽

Immunohistochemical Analysis ◽

Secretory Phase ◽

Normal Endometrium ◽

Mesenchymal Transition ◽

Expression Levels ◽

Endometrial Cells

Endometrial cells and microenvironment are two important factors in the pathogenesis of adenomyosis. Our previous study demonstrated that macrophages can induce eutopic epithelial cells of adenomyosis to suffer from epithelial–mesenchymal transition (EMT). The aim of this study is to detect whether macrophages interacting with epithelial cells equally induce the EMT process in normal and eutopic endometria of healthy and adenomyotic patients; and whether macrophages parallelly polarize to M2. We investigated the expression levels of epithelial cadherin (E-cadherin), neural cadherin (N-cadherin), cytokeratin7 (CK7), vimentin, transforming growth factor-β1 (TGFB1), SMAD3 and pSMAD3 using immunohistochemistry and western blot, and then estimated the genetic levels of CD163, IL10 and MMP12 using real-time quantitative polymerase chain reaction (RT-PCR) in macrophages. Eutopic and normal endometrial tissues were obtained from 20 patients with adenomyosis and 11 control patients without adenomyosis, respectively. The immunohistochemical analysis shows distinct EMT in eutopic endometria in secretory phase; the expression levels of TGFB1, SMAD3 and pSMAD3 that indicate signal pathway of EMT were also higher in secretory phase. Macrophages can induce EMT process in primary endometrial epithelial cells derived from normal and eutopic endometria. After co-culturing, THP-1-derived macrophages polarized to M2. Compared with the eutopic endometrium group, further polarization to M2 was observed in the normal endometrium group. These results indicate that adenomyosis may be promoted by the pathologic EMT of epithelial cells, which is induced by macrophages that incapably polarize to M2.

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FGFR-TKI resistance in cancer: current status and perspectives

Journal of Hematology & Oncology ◽

10.1186/s13045-021-01040-2 ◽

2021 ◽

Vol 14 (1) ◽

Cited By ~ 1

Author(s):

Sitong Yue ◽

Yukun Li ◽

Xiaojuan Chen ◽

Juan Wang ◽

Meixiang Li ◽

...

Keyword(s):

Gene Fusion ◽

Kinase Inhibitors ◽

Current Status ◽

Great Success ◽

Fibroblast Growth Factor Receptors ◽

Pathway Activation ◽

Tki Resistance ◽

Covalent Inhibitors ◽

And Migration ◽

Dual Target

AbstractFibroblast growth factor receptors (FGFRs) play key roles in promoting the proliferation, differentiation, and migration of cancer cell. Inactivation of FGFRs by tyrosine kinase inhibitors (TKI) has achieved great success in tumor-targeted therapy. However, resistance to FGFR-TKI has become a concern. Here, we review the mechanisms of FGFR-TKI resistance in cancer, including gatekeeper mutations, alternative signaling pathway activation, lysosome-mediated TKI sequestration, and gene fusion. In addition, we summarize strategies to overcome resistance, including developing covalent inhibitors, developing dual-target inhibitors, adopting combination therapy, and targeting lysosomes, which will facilitate the transition to precision medicine and individualized treatment.

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Bu Shen Yang Xue Prescription Has Treating Effect on Endometrial Cancer through FSH/PI3K/AKT/Gankyrin/HIF-α/cyclinD 1 Pathway in Ishikawa Cells

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2018/8412984 ◽

2018 ◽

Vol 2018 ◽

pp. 1-10

Author(s):

Yue-qun Chen ◽

Hua-li Fei ◽

Hong-li Zhu

Keyword(s):

Endometrial Cancer ◽

Endometrial Carcinoma ◽

Cancer Cell ◽

Nude Mice ◽

Follicle Stimulating Hormone ◽

Control Group ◽

Model Group ◽

Migration Ability ◽

Ishikawa Cells ◽

And Migration

Background. The formulation of Bu Shen Yang Xue (BSYX) has been clinically used in treating gynecologic disease in China, especially for the development of the endometrium. Endometrial carcinoma is the most common malignant tumor of the female genital tract in developed countries. And few studies have been reported on the antitumor activity of BSYX. Therefore, this study aimed to investigate the effect of BSYX on endometrial cancer and make an initial discussion of the underlining mechanisms in Ishikawa cells. Methods and Results. Firstly, 60 SPF female nude mice were randomly divided into control group, model group, BSYX group, and positive group. The models of subcutaneous tumor xenograft of nude mice were established by injection of human endometrial carcinoma cell line Ishikawa tumor cell suspension. Compared with model group, BSYX reduced effectively tumor volume and changed pathological feature in mice tumor issue. Meanwhile, proteins from tumor issues were detected by western blot analysis. The protein levels of follicle-stimulating hormone receptor (FSHR), p-Akt/Akt, Gankyrin, and cyclinD1 in the model group were higher than those in control group but the expression in BSYX group was lower than that in the model group. The hypoxia inducible factor alpha (HIF-α) protein level in the model group was lower than those in control group and upregulated in BSYX group. In addition, Ishikawa cells were cultured and then exposed to follicle-stimulating hormone (FSH), LY294002, a highly selective PI3K inhibitor and serum containing BSYX, respectively. LY294002 and BSYX markedly decreased the cancer cell viability and migration ability and increased the apoptosis rate. FSH promoted the cancer cell ability and migration ability. LY294002 and BSYX evidently downregulated the proteins levels of FSHR, p-Akt/Akt, Gankyrin, and cyclinD1 and upregulated the expression of HIF-α protein, and FSH was on the opposite. Conclusions. Taken together, our results showed that the formulation of BSYX had antitumor effect on endometrial cancer in vivo and in vitro and was related with FSH/PI3K/AKT/Gankyrin/HIF-α/cyclinD1 transduction pathway.

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Lipocalin 2 induces the epithelial–mesenchymal transition in stressed endometrial epithelial cells: possible correlation with endometriosis development in a mouse model

Reproduction ◽

10.1530/rep-13-0236 ◽

2014 ◽

Vol 147 (2) ◽

pp. 179-187 ◽

Cited By ~ 17

Author(s):

Chi-Jr Liao ◽

Pei-Tzu Li ◽

Ying-Chu Lee ◽

Sheng-Hsiang Li ◽

Sin Tak Chu

Keyword(s):

Epithelial Cells ◽

Mouse Model ◽

Epithelial Mesenchymal Transition ◽

Migration And Invasion ◽

Lipocalin 2 ◽

Mesenchymal Transition ◽

Endometrial Cells ◽

Cellular Microenvironments ◽

And Migration ◽

Endometrial Epithelial Cells

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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Mitonucleons formed during differentiation of Ishikawa endometrial cells generate vacuoles that elevate monolayer syncytia: Differentiation of Ishikawa domes, Part 1

10.7287/peerj.preprints.1728v1 ◽

2016 ◽

Author(s):

Honoree Fleming

Keyword(s):

Cell Death ◽

Structural Changes ◽

Apical Membrane ◽

Signet Ring Cells ◽

Double Membrane ◽

Endometrial Cells ◽

Endometrial Epithelial Cell ◽

Signet Ring ◽

Endogenous Biotin ◽

Cell Profile

In 1998, we published a paper (Fleming et.al, 1998) describing some aspects of Ishikawa endometrial epithelial cell differentiation from monolayer cells into cells forming fluid-filled hemispheres called domes. The process begins with the dissolution of membranes within discrete regions of the monolayer. Nuclei from fused cells aggregate and endogenous biotin in particulate structures assumed to be mitochondria increase throughout the resulting syncytium. Endogenous biotin is also the distinguishing feature of a membrane that surrounds aggregates of multiple nuclei in a structure called a mitonucleon. The current paper includes additional observations on structural changes accompanying Ishikawa differentiation. Vacuoles form in the heterochromatin of the mitonucleon and within the biotin-containing double membrane surrounding heterochromatin. With the formation of vacuoles, the mitonucleon can be seen to rise along with the apical membrane of the syncytium in which it formed. The small vacuoles that form within the heterochromatin result in structures similar to “cells with optically clear nuclei” found in some cancers. The second larger vacuole that forms within the membrane surrounding the heterochromatin transforms the cell profile to one that resembles “signet ring” cells also observed in some cancers. Eventually the membrane surrounding the massed heterochromatin, generated three to four hours earlier, is breached and previously aggregated nuclei disaggregate. During this process heterochromatin in the mitonucleons undergoes changes usually ascribed to cells undergoing programmed cell death such as pyknosis and DNA fragmentation (Fleming, 2016b). The cells do not die, instead chromatin filaments appear to coalesce into a chromatin mass that gives rise to dome-filling nuclei by amitosis during the final three to four hours of the 20 hour differentiation (Fleming, 2016c).

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Pathogenic Effects of Mineralocorticoid Pathway Activation in Retinal Pigment Epithelium

International Journal of Molecular Sciences ◽

10.3390/ijms22179618 ◽

2021 ◽

Vol 22 (17) ◽

pp. 9618

Author(s):

Jérémie Canonica ◽

Min Zhao ◽

Tatiana Favez ◽

Emmanuelle Gelizé ◽

Laurent Jonet ◽

...

Keyword(s):

Extracellular Matrix ◽

Retinal Pigment Epithelium ◽

Barrier Function ◽

Pigment Epithelium ◽

Retinal Pigment ◽

Retinal Diseases ◽

Mesenchymal Transition ◽

Rpe Cells ◽

Pathway Activation ◽

And Migration

Glucocorticoids are amongst the most used drugs to treat retinal diseases of various origins. Yet, the transcriptional regulations induced by glucocorticoid receptor (GR) and mineralocorticoid receptor (MR) activation in retinal pigment epithelium cells (RPE) that form the outer blood–retina barrier are unknown. Levels of endogenous corticoids, ligands for MR and GR, were measured in human ocular media. Human RPE cells derived from induced pluripotent stem cells (iRPE) were used to analyze the pan-transcriptional regulations induced by aldosterone—an MR-specific agonist, or cortisol or cortisol + RU486—a GR antagonist. The retinal phenotype of transgenic mice that overexpress the human MR (P1.hMR) was analyzed. In the human eye, the main ligand for GR and MR is cortisol. The iRPE cells express functional GR and MR. The subset of genes regulated by aldosterone and by cortisol + RU-486, and not by cortisol alone, mimics an imbalance toward MR activation. They are involved in extracellular matrix remodeling (CNN1, MGP, AMTN), epithelial–mesenchymal transition, RPE cell proliferation and migration (ITGB3, PLAUR and FOSL1) and immune balance (TNFSF18 and PTX3). The P1.hMR mice showed choroidal vasodilation, focal alteration of the RPE/choroid interface and migration of RPE cells together with RPE barrier function alteration, similar to human retinal diseases within the pachychoroid spectrum. RPE is a corticosteroid-sensitive epithelium. MR pathway activation in the RPE regulates genes involved in barrier function, extracellular matrix, neural regulation and epithelial differentiation, which could contribute to retinal pathology.

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cdc37 is essential for JNK pathway activation and wound closure in Drosophila

Molecular Biology of the Cell ◽

10.1091/mbc.e18-12-0822 ◽

2019 ◽

Vol 30 (21) ◽

pp. 2651-2658

Author(s):

Chan-wool Lee ◽

Young-Chang Kwon ◽

Youngbin Lee ◽

Min-Yoon Park ◽

Kwang-Min Choe

Keyword(s):

Cell Growth ◽

Wound Closure ◽

Cell Growth And Migration ◽

Jnk Pathway ◽

Protein Levels ◽

Pathway Activation ◽

And Migration ◽

The Stability ◽

Jnk Activation

Wound closure in the Drosophila larval epidermis mainly involves nonproliferative, endocyling epithelial cells. Consequently, it is largely mediated by cell growth and migration. We discovered that both cell growth and migration in Drosophila require the cochaperone-encoding gene cdc37. Larvae lacking cdc37 in the epidermis failed to close wounds, and the cells of the epidermis failed to change cell shape and polarize. Likewise, wound-induced cell growth was significantly reduced, and correlated with a reduction in the size of the cell nucleus. The c-Jun N-terminal kinase (JNK) pathway, which is essential for wound closure, was not typically activated in injured cdc37 knockdown larvae. In addition, JNK, Hep, Mkk4, and Tak1 protein levels were reduced, consistent with previous reports showing that Cdc37 is important for the stability of various client kinases. Protein levels of the integrin β subunit and its wound-induced protein expression were also reduced, reflecting the disruption of JNK activation, which is crucial for expression of integrin β during wound closure. These results are consistent with a role of Cdc37 in maintaining the stability of the JNK pathway kinases, thus mediating cell growth and migration during Drosophila wound healing.

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Endostatin: A Promising Drug for Antiangiogenic Therapy

The International Journal of Biological Markers ◽

10.1177/172460089901400412 ◽

1999 ◽

Vol 14 (4) ◽

pp. 263-267 ◽

Cited By ~ 13

Author(s):

L. Cirri ◽

S. Donnini ◽

L. Morbidelli ◽

P. Chiarugi ◽

M. Ziche ◽

...

Keyword(s):

Cell Proliferation ◽

Endothelial Cell ◽

Growth Factor ◽

Antiangiogenic Therapy ◽

Specific Inhibitor ◽

Angiogenesis Inhibitors ◽

Endothelial Cell Proliferation ◽

Platelet Factor ◽

Collagen Xviii ◽

And Migration

Angiogenesis, the formation of new blood vessels from existing capillaries, is critical for tumors to grow beyond a few in size. Tumor cells produce one or more angiogenic factors including fibroblast growth factor and vascular endothelial growth factor. Surprisingly, antiangiogenic factors or angiogenesis inhibitors have been isolated from tumors. Some angiogenesis inhibitors, such as angiostatin, are associated with tumors while others, such as platelet-factor 4 and interferon-alpha are not. Endostatin, a C-terminal product of collagen XVIII, is a specific inhibitor of endothelial cell proliferation, migration and angiogenesis. The mechanism by which endostatin inhibits endothelial cell proliferation and migration is unknown. Endostatin was originally expressed in a prokaryotic system and, late, in a yeast system, thanks to which it is possible to obtain a sufficient quantity of the protein in a soluble and refolded form to be used in preclincial and clinical trials.

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Estrogen signaling in the proliferative endometrium: implications in endometriosis

Revista da Associação Médica Brasileira ◽

10.1590/1806-9282.62.01.72 ◽

2016 ◽

Vol 62 (1) ◽

pp. 72-77 ◽

Cited By ~ 10

Author(s):

Rita de Cássia Pereira da Costa e Silva ◽

Kátia Karina Verolli de Oliveira Moura ◽

Circoncisto Laurentino Ribeiro Júnior ◽

Lidia Andreu Guillo

Keyword(s):

Signaling Pathways ◽

Physiological Role ◽

Estrogen Signaling ◽

Normal Endometrium ◽

Cell Culture Techniques ◽

Culture Techniques ◽

Endometrial Cells ◽

Signaling Mechanisms ◽

Proliferative Endometrium

SUMMARY Even though the physiological role of estrogen in the female reproductive cycle and endometrial proliferative phase is well established, the signaling pathways by which estrogen exerts its action in the endometrial tissue are still little known. In this regard, advancements in cell culture techniques and maintenance of endometrial cells in cultures enabled the discovery of new signaling mechanisms activated by estrogen in the normal endometrium and in endometriosis. This review aims to present the recent findings in the genomic and non-genomic estrogen signaling pathways in the proliferative human endometrium specifically associated with the pathogenesis and development of endometriosis.

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