Increased expression of CYP1A1 and CYP1B1 in ovarian/peritoneal endometriotic lesions

Abstract Endometriosis is an estrogen-dependent disease affecting up to 10% of all premenopausal women. There is evidence that different endometriosis sites show distinct local estrogen concentration, which, in turn, might be due to a unique local estrogen metabolism. We aimed to investigate whether there was a site-specific regulation of selected enzymes responsible for the oxidative metabolism of estrogens in biopsy samples and endometrial and endometriotic stromal cells. Cytochrome P450 (CYP) 1A1 and CYP1B1 mRNA and protein expressions in deep-infiltrating (rectal, retossigmoidal, and uterossacral) lesions, superficial (ovarian and peritoneal) lesions, and eutopic and healthy (control) endometrium were evaluated by real-time PCR and western blot. Using a cross-sectional study design with 58 premenopausal women who were not under hormonal treatment, we were able to identify an overall increased CYP1A1 and CYP1B1 mRNA expression in superficial lesions compared with the healthy endometrium. CYP1A1 mRNA expression in superficial lesions was also greater than in the eutopic endometrium. Interestingly, we found a similar pattern of CYP1A1 and CYP1B1 expression in in vitro stromal cells isolated from ovarian lesions (n=3) when compared with stromal cells isolated from either rectum lesions or eutopic endometrium. In contradiction, there was an increased half-life of estradiol (measured by HPLC-MS-MS) in ovarian endometriotic stromal cells compared with paired eutopic stromal endometrial cells. Our results indicate that there is a site-dependent regulation of CYP1A1 and CYP1B1 in ovarian/peritoneal lesions and ovarian endometriotic stromal cells, whereas a slower metabolism is taking place in these cells.

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The expression of BECN1, LC3B, and BCL2 genes in eutopic endometrium of patients with adenomyosis: A cross-sectional study

Journal of Endometriosis and Pelvic Pain Disorders ◽

10.1177/22840265211046750 ◽

2021 ◽

pp. 228402652110467

Author(s):

Siriluk Tantanavipas ◽

Tepchongchit Aojanepong ◽

Wanwisa Waiyaput ◽

Areepan Sophonsritsuk

Keyword(s):

Mrna Expression ◽

Geometric Mean ◽

Premenopausal Women ◽

Cross Sectional Study ◽

Control Group ◽

Sectional Study ◽

Menstrual Phase ◽

Cross Sectional ◽

Eutopic Endometrium ◽

Significant Difference

Introduction: Both autophagy and apoptosis play a role in the cyclic remodeling of the endometrium. The abnormal regulation of genes and signaling pathways in the eutopic endometrium plays a role in the abnormal migration and implantation in adenomyosis. Objective: The present study investigates the mRNA expression of autophagy and apoptosis-related genes BECN1, LC3B, and BCL2 in the eutopic endometrium of patients with adenomyosis compared with healthy premenopausal women. Materials and methods: The present work was a cross-sectional study conducted between July 2018 and April 2019. The participants were 32 premenopausal women who attended the surgery for adenomyosis and other benign gynecological conditions. The participants were divided into two groups, with 16 women in the adenomyosis group and 16 healthy women in the control group. Endometrial tissues were collected during the proliferative menstrual phase for a quantitative real-time polymerase chain reaction. Results: The mRNA expression of BECN1, LC3B, and BCL2 were normalized by geometric mean mRNA expression of actin and GAPDH. There was no significant difference in mRNA expression for all three genes when comparing the control and adenomyosis groups. Conclusions: The mRNA expressions of autophagy-related genes BECN1 and LC3B and anti-apoptosis-related gene BCL2 were not significantly different in the eutopic endometrium of patients with adenomyosis compared with healthy premenopausal women during the proliferative menstrual phase.

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Evidence for a PGF2α auto-amplification system in the endometrium in mares

Reproduction ◽

10.1530/rep-15-0617 ◽

2016 ◽

Vol 151 (5) ◽

pp. 517-526 ◽

Cited By ~ 6

Author(s):

Keisuke Kozai ◽

Shota Tokuyama ◽

Anna Z Szóstek ◽

Yuko Toishi ◽

Nobuo Tsunoda ◽

...

Keyword(s):

Mrna Expression ◽

Stromal Cells ◽

Luteal Phase ◽

Endometrial Cell ◽

Late Luteal Phase ◽

Prostaglandin F ◽

Amplification System ◽

Prostaglandin Endoperoxide Synthase

AbstractIn mares, prostaglandin F2α(PGF2α) secreted from the endometrium is a major luteolysin. Some domestic animals have an auto-amplification system in which PGF2αcan stimulate its own production. Here, we investigated whether this is also the case in mares. In anin vivostudy, mares at the mid-luteal phase (days 6–8 of estrous cycle) were injected i.m. with cloprostenol (250 µg) and blood samples were collected at fixed intervals until 72 h after treatment. Progesterone (P4) concentrations started decreasing 45 min after the injection and continued to decrease up to 24 h (P < 0.05). In turn, 13,14-dihydro-15-keto-PGF2α(PGFM) metabolite started to increase 4h after an injection and continued to increase up to 72 h (P < 0.05). PGF receptor (PTGFR) mRNA expression in the endometrium was significantly higher in the late luteal phase than in the early and regressed luteal phases (P < 0.05).In vitro, PGF2αsignificantly stimulated (P < 0.05) PGF2αproduction by endometrial tissues and endometrial epithelial and stromal cells and significantly increased (P < 0.05) the mRNA expression of prostaglandin-endoperoxide synthase-2 (PTGS2), an enzyme involved in PGF2αsynthesis in endometrial cell. These findings strongly suggest the existence of an endometrial PGF2αauto-amplification system in mares.

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Indoleamine 2,3-dioxygenase suppresses the cytotoxicity of 1 NK cells in response to ectopic endometrial stromal cells in endometriosis

Reproduction ◽

10.1530/rep-18-0112 ◽

2018 ◽

Cited By ~ 1

Author(s):

Xuan-Tong Liu ◽

Hui-Ting Sun ◽

Zhong-Fang Zhang ◽

Ru-Xia Shi ◽

Li-Bing Liu ◽

...

Keyword(s):

Nk Cells ◽

Stromal Cells ◽

Therapeutic Strategy ◽

Neutralizing Antibody ◽

Inhibitory Effect ◽

Endometrial Stromal Cells ◽

Endometrial Cells ◽

Indoleamine 2,3 Dioxygenase ◽

Healthy Control

It has been reported that the impaired cytotoxicity of natural killer (NK) cells and abnormal cytokines that are changed by the interaction between ectopic endometrial cells and immune cells is indispensable for the initiation and development of endometriosis (EMS). However, the mechanism of NK cells dysfunction in EMS remains largely unclear. Here, we found that NK cells in peritoneal fluid from women with EMS highly expressed indoleamine 2,3-dioxygenase (IDO). Furthermore, IDO+NK cells possessed lower NKp46 and NKG2D but higher IL-10 than that of IDO-NK. Co-culture with endometrial stromal cells (nESCs) from healthy control or ectopic ESCs (eESCs) from women with EMS led to a significant increase in the IDO level in NK cells from peripheral blood, particularly eESCs, and an anti-TGF-β neutralizing antibody suppressed these effects in vitro. NK cells co-cultured with ESC more preferentially inhibited the viability of nESCs than eESCs did, and pretreating with 1-methyl-tryptophan (1-MT), an IDO inhibitor, reversed the inhibitory effect of NK cells on eESC viability. These data suggest that ESCs induce IDO+NK cells differentiation partly by TGF-β, and that IDO further restricts the cytotoxicity of NK cells in response to eESCs, which provides a potential therapeutic strategy for EMS patients, particularly those with a high number of impaired cytotoxic IDO+NK cells.

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Decorin induced by progesterone plays a crucial role in suppressing endometriosis

Journal of Endocrinology ◽

10.1530/joe-14-0393 ◽

2014 ◽

Vol 223 (2) ◽

pp. 203-216 ◽

Cited By ~ 13

Author(s):

Yoshihiro Joshua Ono ◽

Yoshito Terai ◽

Akiko Tanabe ◽

Atsushi Hayashi ◽

Masami Hayashi ◽

...

Keyword(s):

Cell Cycle ◽

Mrna Expression ◽

Cell Lines ◽

Stromal Cells ◽

Crucial Role ◽

Dependent Manner ◽

Endometrial Stromal Cells ◽

Endometrial Cells ◽

Cycle Arrest ◽

Dose Dependent

Dienogest, a synthetic progestin, has been shown to be effective against endometriosis, although it is still unclear as to how it affects the ectopic endometrial cells. Decorin has been shown to be a powerful endogenous tumor repressor acting in a paracrine fashion to limit tumor growth. Our objectives were to examine the direct effects of progesterone and dienogest on the in vitro proliferation of the human ectopic endometrial epithelial and stromal cell lines, and evaluate as to how decorin contributes to this effect. We also examined DCN mRNA expression in 50 endometriosis patients. The growth of both cell lines was inhibited in a dose-dependent manner by both decorin and dienogest. Using a chromatin immunoprecipitation assay, it was noted that progesterone and dienogest directly induced the binding of the decorin promoter in the EMOsis cc/TERT cells (immortalized human ovarian epithelial cells) and CRL-4003 cells (immortalized human endometrial stromal cells). Progesterone and dienogest also led to significant induced cell cycle arrest via decorin by promoting production of p21 in both cell lines in a dose-dependent manner. Decorin also suppressed the expression of MET in both cell lines. We confirmed that DCN mRNA expression in patients treated with dienogest was higher than that in the control group. In conclusion, decorin induced by dienogest appears to play a crucial role in suppressing endometriosis by exerting anti-proliferative effects and inducing cell cycle arrest via the production of p21 human ectopic endometrial cells and eutopic endometrial stromal cells.

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Ovarian steroids affect prostaglandin production in equine endometrial cells in vitro

Journal of Endocrinology ◽

10.1530/joe-13-0185 ◽

2014 ◽

Vol 220 (3) ◽

pp. 263-276 ◽

Cited By ~ 12

Author(s):

Anna Z Szóstek ◽

António M Galvão ◽

Graça M Ferreira-Dias ◽

Dariusz J Skarzynski

Keyword(s):

Cell Proliferation ◽

Epithelial Cells ◽

Protein Expression ◽

Stromal Cells ◽

Positive Control ◽

Dependent Manner ◽

Ovarian Steroids ◽

Endometrial Cells ◽

Prostaglandin Endoperoxide Synthase

This study aimed to evaluate the influence of ovarian steroids on equine endometrial epithelial and stromal cells, specifically i) prostaglandin (PG) production in a time-dependent manner, ii) specific PG synthases mRNA transcription and protein expression, and iii) cell proliferation. After passage I, cells were exposed to vehicle, oxytocin (OT, positive control, 10−7M), progesterone (P4, 10−7M), 17β estradiol (E2, 10−9M), or P4+E2for 12, 24, 48, or 72 h. Following treatment, PG concentration was determined using the direct enzyme immunoassay (EIA) method. Alterations inPGsynthases mRNA transcriptions,PGsynthases protein expression, and cell proliferation in response to the treatments were determined after 24 h using real-time PCR, western blot, or 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide respectively. After 24 h, E2and P4+E2increased PGE2and PGF2αsecretion as well as specific prostaglandin-endoperoxide synthase-2 (PTGS2), PGE2synthases (PGES), and PGF2αsynthases (PGFS) expression in the epithelial cells (P<0.05). Additionally, E2and P4+E2increased PTGS2 expression in stromal cells after 24 h (P<0.05). In stromal cells, P4+E2increased PGE2production as well as PGES expression after 24 h (P<0.05). Both E2and P4+E2increased PGF2αproduction by stromal cells after 24 h (P<0.05). Ovarian steroids affected proliferation of stromal and epithelial cells during the 24-h incubation period (P<0.05). We provide evidence that ovarian steroids affect PG production in equine endometrial cells, upregulating PTGS2, PGES, and PGFS expression. Ovarian steroid-stimulated PG production could be an important mechanism occurring in the equine endometrium that is involved in the regulation of the estrous cycle and early pregnancy.

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Immunosuppressive Factor MNSFβ Regulates Cytokine Secretion by Mouse Lymphocytes and Is Involved in Interactions between the Mouse Embryo and Endometrial Cells In Vitro

ISRN Immunology ◽

10.5402/2011/186541 ◽

2011 ◽

Vol 2011 ◽

pp. 1-11 ◽

Cited By ~ 2

Author(s):

Yaping He ◽

Zhaogui Sun ◽

Yan Shi ◽

Yahong Jiang ◽

Zhefu Jia ◽

...

Keyword(s):

Stromal Cells ◽

Embryo Implantation ◽

Cytokine Secretion ◽

Embryonic Cells ◽

Suppressor Factor ◽

Endometrial Stromal Cells ◽

Endometrial Cells ◽

Coculture Model ◽

Mouse Lymphocytes

Immune tolerance at the fetomaternal interface must be established during the processes of implantation and pregnancy. Monoclonal nonspecific suppressor factor beta (MNSFβ) is a secreted protein that possesses antigen-nonspecific immune-suppressive function. It was previously reported that intrauterine immunoneutralization of MNSFβ significantly inhibited embryo implantation in mice. In the present study, MNSFβ protein expression was up- or downregulated by overexpression or RNA interference, respectively, in HCC-94 cells and the culture supernatants used to determine effects of MNSFβ on the secretion of IL-4 and TNFα from mouse lymphocytes as detected by ELISA. A coculture model of mouse embryos and endometrial stromal cells was also utilized to determine the effects of a specific anti-MNSFβ antibody on hatching and growth of embryos in vitro. The results show that MNSFβ induced secretion of IL-4 and inhibited secretion of TNFα from mouse lymphocytes. Following immunoneutralization of MNSFβ protein in the HCC-94 supernatant, the stimulatory effect of MNSFβ on IL-4 secretion from mouse lymphocytes was reduced, while the inhibitory effect on secretion of TNFα was abrogated. Expression of MNSFβ was detected in both embryonic and endometrial stromal cells, and its immunoneutralization inhibited the hatching and spreading of embryos in an in vitro coculture model. These results indicated that MNSFβ may play critical roles during the peri-implantation process by regulating cytokine secretion of lymphocytes and by mediating the crosstalk between embryonic cells and endometrial stromal cells.

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Mesenchymal Stromal Cells Support Endometriotic Stromal CellsIn Vitro

Stem Cells International ◽

10.1155/2018/7318513 ◽

2018 ◽

Vol 2018 ◽

pp. 1-12 ◽

Cited By ~ 2

Author(s):

Fawaz Abomaray ◽

Sebastian Gidlöf ◽

Bartosz Bezubik ◽

Mikael Engman ◽

Cecilia Götherström

Keyword(s):

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Inflammatory Diseases ◽

Umbilical Vein ◽

Cell Types ◽

Tube Formation ◽

Migration And Invasion ◽

Endometrial Cells ◽

Umbilical Vein Endothelial Cells

Endometriosis is an inflammatory disease marked by ectopic growth of endometrial cells. Mesenchymal stromal cells (MSC) have immunosuppressive properties that have been suggested as a treatment for inflammatory diseases. Therefore, the aim herein was to examine effects of allogeneic MSC on endometriosis-derived cellsin vitroas a potential therapy for endometriosis. MSC from allogeneic adipose tissue (Ad-MSC) and stromal cells from endometrium (ESCendo) and endometriotic ovarian cysts (ESCcyst) from women with endometriosis were isolated. The effects of Ad-MSC on ESCendoand ESCcystwere investigated usingin vitroproliferation, apoptosis, adhesion, tube formation, migration, and invasion assays. Ad-MSC significantly increased proliferation of ESC compared to untreated controls. Moreover, Ad-MSC significantly decreased apoptosis and increased survival of ESC. Ad-MSC significantly increased adhesion of ESCendoand not ESCcyston fibronectin. Conditioned medium from cocultures of Ad-MSC and ESC significantly increased tube formation of human umbilical vein endothelial cells on matrigel. Ad-MSC may significantly increase migration of ESCcystand did not increase invasion of both cell types. The data suggest that allogeneic Ad-MSC should not be considered as a potential therapy for endometriosis, because they may support the pathology by maintaining and increasing growth of ectopic endometrial tissue.

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Changes in the extracellular microenvironment and osteogenic responses of mesenchymal stem/stromal cells induced by in vitro direct electrical stimulation

Journal of Tissue Engineering ◽

10.1177/2041731420974147 ◽

2021 ◽

Vol 12 ◽

pp. 204173142097414

Author(s):

Kasama Srirussamee ◽

Ruikang Xue ◽

Sahba Mobini ◽

Nigel J Cassidy ◽

Sarah H Cartmell

Keyword(s):

Electrical Stimulation ◽

Mrna Expression ◽

Stromal Cells ◽

Control Cell ◽

Cell Monolayer ◽

Computational Modelling ◽

Bone Injury ◽

Metabolic Activities ◽

By Products

Electrical stimulation (ES) has potential to be an effective tool for bone injury treatment in clinics. However, the therapeutic mechanism associated with ES is still being discussed. This study aims to investigate the initial mechanism of action by characterising the physical and chemical changes in the extracellular environment during ES and correlate them with the responses of mesenchymal stem/stromal cells (MSCs). Computational modelling was used to estimate the electrical potentials relative to the cathode and the current density across the cell monolayer. We showed expression of phosphorylated ERK1/2, c-FOS, c-JUN, and SPP1 mRNAs, as well as the increased metabolic activities of MSCs at different time points. Moreover, the average of 2.5 μM of H2O2 and 34 μg/L of dissolved Pt were measured from the electrically stimulated media (ES media), which also corresponded with the increases in SPP1 mRNA expression and cell metabolic activities. The addition of sodium pyruvate to the ES media as an antioxidant did not alter the SPP1 mRNA expression, but eliminated an increase in cell metabolic activities induced by ES media treatment. These findings suggest that H2O2 was influencing cell metabolic activity, whereas SPP1 mRNA expression was regulated by other faradic by-products. This study reveals how different electrical stimulation regime alters cellular regenerative responses and the roles of faradic by-products, that might be used as a physical tool to guide and control cell behaviour.

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Mesenchymal Stromal Cells Isolated from Ectopic but Not Eutopic Endometrium Display Pronounced Immunomodulatory Activity In Vitro

Biomedicines ◽

10.3390/biomedicines9101286 ◽

2021 ◽

Vol 9 (10) ◽

pp. 1286

Author(s):

Alexey Yu. Lupatov ◽

Roza Yu. Saryglar ◽

Valentina V. Vtorushina ◽

Rimma A. Poltavtseva ◽

Oxana A. Bystrykh ◽

...

Keyword(s):

Cell Cultures ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Immunomodulatory Activity ◽

Normal Endometrium ◽

Eutopic Endometrium ◽

Dendritic Cell Differentiation ◽

Extragenital Endometriosis ◽

Similar Phenotype

A comparative analysis of the cell surface markers and immunological properties of cell cultures originating from normal endometrium and endometrioid heterotopias of women with extragenital endometriosis was carried out. Both types of cell cultures expressed surface molecules typical of mesenchymal stromal cells and did not express hematopoietic and epithelial markers. Despite similar phenotype, the mesenchymal stromal cells derived from the two sources had different immunomodulation capacities: the cells of endometrioid heterotopias but not eutopic endometrium could suppress dendritic cell differentiation from monocytes as well as lymphocyte proliferation in allogeneic co-cultures. A comparative multiplex analysis of the secretomes revealed a significant increase in the secretion of pro-inflammatory mediators, including IL6, IFN-γ, and several chemokines associated with inflammation by the stromal cells of ectopic lesions. The results demonstrate that the stromal cells of endometrioid heterotopias display enhanced pro-inflammatory and immunosuppressive activities, which most likely impact the pathogenesis and progression of the disease.

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The scar constituent Collagen I triggers coordinated collective migration and invasion in a 3D spheroid model of early endometriotic lesions

10.1101/2020.04.02.005322 ◽

2020 ◽

Author(s):

Anna Stejskalova ◽

Victoria Fincke ◽

Melissa Nowak ◽

Yvonne Schmidt ◽

Marie-Kristin von Wahlde ◽

...

Keyword(s):

Stromal Cells ◽

3D Model ◽

Single Cells ◽

Experimental Models ◽

Collagen I ◽

Migration And Invasion ◽

Collective Migration ◽

Endometrial Stromal Cells ◽

Endometrial Cells

AbstractEndometriosis is a painful gynaecological condition characterized by ectopic growth of endometrial cells outside of the uterus. Little is known about the mechanisms by which endometrial fragments invade tissues. This is partially due to a lack of suitable experimental models. In this study, we show that a spheroid 3D model, but not single cells mimic the collective endometrial fragment-like invasion through the extracellular matrix. This model reveals that collagen I, the main constituent of surgical scars, significantly increases the rate of lesion formation by healthy endometrial stromal cells (St-T1b) in vitro compared to the basement membrane-like matrix Matrigel. Stromal cell invasion of collagen I requires MMPs, whereas collective migration of endometriotic epithelial 12Z cells involves Rac-signalling. We show that inhibiting ROCK signalling responsible for actomyosin contraction increases the lesion-size. Moreover, endometriotic epithelial 12Z cells, but not eutopic stromal cells St-T1b migrate on Matrigel. The rate of this migration is decreased by the microRNA miR-200b and increased by miR-145. Our 3D model offers a facile approach to dissect how endometrial fragments invade tissues and is an important step toward developing new personalized therapeutics for endometriosis. Moreover, our model is a suitable tool to screen small molecule drugs and microRNA-based therapeutics.

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