scholarly journals Expression of Progesterone Receptor A form and Its Role in the Interaction of Progesterone with Cortisol on Cyclooxygenase-2 Expression in Amnionic Fibroblasts

2009 ◽  
Vol 94 (12) ◽  
pp. 5085-5092 ◽  
Author(s):  
C. M. Guo ◽  
X. O. Zhu ◽  
X. T. Ni ◽  
Z. Yang ◽  
L. Myatt ◽  
...  
Keyword(s):  
Progesterone Receptor ◽  
Small Interfering Rna ◽  
Prostaglandin Synthesis ◽  
Cyclooxygenase 2 ◽  
Receptor Subtypes ◽  
Term Pregnancy ◽  
Human Amnion ◽  
Cox 2 ◽  
Exogenous Progesterone ◽  
Interfering Rna

Context: Human amnion fibroblasts produce abundant prostaglandins toward the end of gestation, which is believed to be one of the major events leading to parturition. Glucocorticoids have been shown to up-regulate cyclooxygenase-2 (COX-2) expression, the crucial enzyme catalyzing prostaglandin synthesis, in human amnion fibroblasts. Although a major propregnancy hormone, the effect of progesterone and the associated progesterone receptor subtypes in the regulation of both basal and glucocorticoid-induced COX-2 expression in human amnion fibroblasts have not been resolved. Methods and Results: Cultured human amnion fibroblasts prepared from the fetal membranes at term pregnancy without labor mainly expressed the progesterone receptor A form (PRA). Inhibition of endogenous progesterone production with trilostane or knockdown of PRA expression with small interfering RNA significantly enhanced the glucocorticoid receptor (GR)-mediated COX-2 induction by cortisol, whereas overexpression of PRA attenuated the induction by cortisol. Co-immunoprecipitation assay revealed PRA in the GR protein complex. Although exogenous progesterone did not alter COX-2 expression under basal conditions, it attenuated cortisol-induced COX-2 expression at concentrations about 10- to 50-fold higher, which might be achieved by competition with cortisol for GR. Conclusions: We demonstrated in this study that endogenous progesterone might counteract the induction of prostaglandin synthesis by cortisol via PRA transdominant repression of GR function, whereas high levels of progesterone might further inhibit the induction by cortisol via competitive binding to GR in human amnion fibroblasts. These inhibitory actions of progesterone and PRA on glucocorticoids and GR may partly explain the inconsistent effects of glucocorticoids on parturition in humans.

scholarly journals Transcriptional Cross Talk between the Forkhead Transcription Factor Forkhead Box O1A and the Progesterone Receptor Coordinates Cell Cycle Regulation and Differentiation in Human Endometrial Stromal Cells

2007 ◽  
Vol 21 (10) ◽  
pp. 2334-2349 ◽  
Author(s):  
Masashi Takano ◽  
Zhenxiao Lu ◽  
Tomoko Goto ◽  
Luca Fusi ◽  
Jenny Higham ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Progesterone Receptor ◽  
Stromal Cells ◽  
Small Interfering Rna ◽  
Cell Cycle Regulation ◽  
Forkhead Transcription Factor ◽  
Endometrial Stromal Cells ◽  
Forkhead Box ◽  
Cycle Regulation ◽  
Interfering Rna

Abstract Differentiation of human endometrial stromal cells (HESCs) into decidual cells is associated with induction of the forkhead transcription factor forkhead box O1A (FOXO1). We performed a genomic screen to identify decidua-specific genes under FOXO1 control. Primary HESCs were transfected with small interfering RNA targeting FOXO1 or with nontargeting control small interfering RNA before treatment with a cAMP analogue and the progestin, medroxyprogesterone acetate for 72 h. Total RNA was processed for whole genome analysis using high-density oligonucleotide arrays. We identified 3405 significantly regulated genes upon decidualization of HESCs, 507 (15.3%) of which were aberrantly expressed upon FOXO1 knockdown. Among the most up-regulated FOXO1-dependent transcriptional targets were WNT signaling-related genes (WNT4, WNT16 ), the insulin receptor (INSR), differentiation markers (PRL, IGFBP1, and LEFTY2), and the cyclin-dependent kinase inhibitor p57Kip2 (CDKN1C). Analysis of FOXO1-dependent down-regulated genes uncovered several factors involved in cell cycle regulation, including CCNB1, CCNB2, MCM5, CDC2 and NEK2. Cell viability assay and cell cycle analysis demonstrated that FOXO1 silencing promotes proliferation of differentiating HESCs. Using a glutathione-S-transferase pull-down assay, we confirmed that FOXO1 interacts with progesterone receptor, irrespectively of the presence of ligand. In agreement, knockdown of PR disrupted the regulation of FOXO1 target genes involved in differentiation (IGFBP1, PRL, and WNT4) and cell cycle regulation (CDKN1, CCNB2 and CDC2) in HESCs treated with either cAMP plus medroxyprogesterone acetate or with cAMP alone. Together, the data demonstrate that FOXO1 engages in transcriptional cross talk with progesterone receptor to coordinate cell cycle regulation and differentiation of HESCs.

scholarly journals Expression of cyclo-oxygenase types-1 and -2 in human fetal membranes throughout pregnancy

1999 ◽  
Vol 22 (2) ◽  
pp. 125-130 ◽  
Author(s):  
D Slater ◽  
W Dennes ◽  
R Sawdy ◽  
V Allport ◽  
P Bennett
Keyword(s):  
Mrna Expression ◽  
Gestational Age ◽  
Prostaglandin Synthesis ◽  
Fetal Membranes ◽  
Activity Levels ◽  
Third Trimester ◽  
Human Amnion ◽  
Human Labour ◽  
Cox 2 ◽  
Cox 1

Human labour is associated with increased prostaglandin synthesis within the fetal membranes. We have studied the expression of the two isoforms of the central prostaglandin synthetic enzyme, cyclo-oxygenase (COX-1 and COX-2), in human fetal membranes throughout pregnancy, at mRNA, protein and activity levels. COX-1 mRNA expression was low in human amnion and chorion-decidua and did not change with gestational age. COX-2 mRNA expression in fetal membranes increased with gestational age, with significant up-regulation prior to the onset of labour and in association with labour. Protein concentrations of COX-1 did not change, whilst concentrations of COX-2 increased from the first to the third trimester. COX activity increased with gestational age and in association with labour, although prostaglandin production in fetal membranes collected after labour was reduced, suggesting reduced substrate supply. These data suggest that it is up-regulation of COX-2, rather than of COX-1, which mediates increased prostaglandin synthesis within the fetal membranes at term. Much of the increase in COX-2 expression precedes the onset of labour, suggesting that it is a cause, rather than a consequence, of labour.

scholarly journals Cross Talk between PKC and CREB in the Induction of COX-2 by PGF2α in Human Amnion Fibroblasts

Endocrinology ◽  
2012 ◽  
Vol 153 (10) ◽  
pp. 4938-4945 ◽  
Author(s):  
C. M. Guo ◽  
N. Kasaraneni ◽  
K. Sun ◽  
L. Myatt
Keyword(s):  
Prostaglandin F2α ◽  
Dominant Negative ◽  
Fetal Membranes ◽  
Camp Response Element ◽  
Human Amnion ◽  
Rate Limiting Step ◽  
Inducible Synthesis ◽  
Fp Receptor ◽  
Cox 2 ◽  
Interfering Rna

Abstract Compelling evidence indicates a crucial role of prostaglandin F2α (PGF2α) in parturition. Both the maternal and fetal sides of the fetal membranes synthesize PGF2α, which exerts effects via the prostaglandin F2α receptor (FP) that is coupled to the activation of protein kinase C (PKC). Cyclooxygenase-2 (COX-2) catalyzes the rate-limiting step of the inducible synthesis of prostaglandin. Although activation of PKC is known to induce COX-2 expression, it is not clear whether PGF2α can induce COX-2 via FP receptor-coupled PKC activation. COX-2 promoter carries a cAMP-response element (CRE) and phosphorylation of CRE binding protein 1 (CREB1) is associated with COX-2 expression in human amnion fibroblasts. We demonstrated that human amnion fibroblasts produced PGF2α and expressed FP receptor. PGF2α increased COX-2 expression and CREB1 phosphorylation, which could be blocked by either the FP receptor antagonist AL8810 or PKC inhibitor Ro31-7549. The PKC activator, phorbol-12-myristate-13-acetate (PMA), could mimic the induction of COX-2 and CREB1 phosphorylation. The induction of COX-2 by PGF2α and PMA could be attenuated by the small interfering RNA-mediated knockdown of CREB1 expression or overexpressing dominant-negative CREB1. A chromatin immunoprecipitation assay showed that the binding of CREB1 to the COX-2 promoter was increased by PGF2α and PMA in amnion fibroblasts. In conclusion, we provide evidence that PGF2α induces COX-2 expression via the FP receptor and phosphorylates CREB1 by PKC, thus increasing CREB1 binding to the COX-2 promoter and the expression of COX-2 in human amnion fibroblasts. This feed-forward loop may be crucial for the production of prostaglandins in the fetal membranes prior to the onset of labor.

scholarly journals Ubiquitin-Proteasomal Degradation of COX-2 in TGF-β Stimulated Human Endometrial Cells Is Mediated Through Endoplasmic Reticulum Mannosidase I

Endocrinology ◽  
2012 ◽  
Vol 153 (1) ◽  
pp. 426-437 ◽  
Author(s):  
Mohan Singh ◽  
Parvesh Chaudhry ◽  
Sophie Parent ◽  
Eric Asselin
Keyword(s):  
Endoplasmic Reticulum ◽  
Stromal Cells ◽  
Small Interfering Rna ◽  
Proteasomal Degradation ◽  
26S Proteasome ◽  
Dependent Manner ◽  
Endometrial Stromal Cells ◽  
Endometrial Cells ◽  
Cox 2 ◽  
Interfering Rna

Cyclooxygenase (COX)-2 is a key regulatory enzyme in the production of prostaglandins (PG) during various physiological processes. Mechanisms of COX-2 regulation in human endometrial stromal cells (human endometrial stromal cells) are not fully understood. In this study, we investigate the role of TGF-β in the regulation of COX-2 in human uterine stromal cells. Each TGF-β isoform decreases COX-2 protein level in human uterine stromal cells in Smad2/3-dependent manner. The decrease in COX-2 is accompanied by a decrease in PG synthesis. Knockdown of Smad4 using specific small interfering RNA prevents the decrease in COX-2 protein, confirming that Smad pathway is implicated in the regulation of COX-2 expression in human endometrial stromal cells. Pretreatment with 26S proteasome inhibitor, MG132, significantly restores COX-2 protein and PG synthesis, indicating that COX-2 undergoes proteasomal degradation in the presence of TGF-β. In addition, each TGF-β isoform up-regulates endoplasmic reticulum (ER)-mannosidase I (ERManI) implying that COX-2 degradation is mediated through ER-associated degradation pathway in these cells. Furthermore, inhibition of ERManI activity using the mannosidase inhibitor (kifunensine), or small interfering RNA-mediated knockdown of ERManI, prevents TGF-β-induced COX-2 degradation. Taken together, these studies suggest that TGF-β promotes COX-2 degradation in a Smad-dependent manner by up-regulating the expression of ERManI and thereby enhancing ER-associated degradation and proteasomal degradation pathways.

scholarly journals Progesterone Receptor Plays a Major Antiinflammatory Role in Human Myometrial Cells by Antagonism of Nuclear Factor-κB Activation of Cyclooxygenase 2 Expression

2006 ◽  
Vol 20 (11) ◽  
pp. 2724-2733 ◽  
Author(s):  
Daniel B. Hardy ◽  
Bethany A. Janowski ◽  
David R. Corey ◽  
Carole R. Mendelson
Keyword(s):  
Progesterone Receptor ◽  
Target Genes ◽  
Nuclear Factor Κb ◽  
Inhibitory Effect ◽  
Cyclooxygenase 2 ◽  
Nuclear Factor ◽  
Uterine Contractility ◽  
T47d Cells ◽  
Myometrial Cells ◽  
Cox 2

Abstract Spontaneous labor in women and in other mammals is likely mediated by a concerted series of biochemical events that negatively impact the ability of the progesterone receptor (PR) to regulate target genes that maintain myometrial quiescence. In the present study, we tested the hypothesis that progesterone/PR inhibits uterine contractility by blocking nuclear factor κB (NF-κB) activation and induction of cyclooxygenase-2 (COX-2), a contractile gene that is up-regulated in labor. To uncover mechanisms for regulation of uterine COX-2, immortalized human fundal myometrial cells were treated with IL-1β ± progesterone. IL-1β alone caused a marked up-regulation of COX-2 mRNA, whereas treatment with progesterone suppressed this induction. This was also observed in human breast cancer (T47D) cells. In both cell lines, this inhibitory effect of progesterone was blocked by RU486. Using chromatin immunoprecipitation, we observed that IL-1β stimulated recruitment of NF-κB p65 to both proximal and distal NF-κB elements of the COX-2 promoter; these effects were diminished by coincubation with progesterone. The ability of progesterone to inhibit COX-2 expression in myometrial cells was associated with rapid induction of mRNA and protein levels of inhibitor of κBα, a protein that blocks NF-κB transactivation. Furthermore, small interfering RNA-mediated ablation of both PR-A and PR-B isoforms in T47D cells greatly enhanced NF-κB activation and COX-2 expression. These effects were observed in the absence of exogenous progesterone, suggesting a ligand-independent action of PR. Based on these findings, we propose that PR may inhibit NF-κB activation of COX-2 gene expression and uterine contractility via ligand-dependent and ligand-independent mechanisms.

Gonadotropin-Induced Apoptosis in Human Ovarian Surface Epithelial Cells Is Associated with Cyclooxygenase-2 Up-Regulation via the β-Catenin/T-Cell Factor Signaling Pathway

2006 ◽  
Vol 20 (12) ◽  
pp. 3336-3350 ◽  
Author(s):  
Yuen Lam Pon ◽  
Alice S. T. Wong
Keyword(s):  
T Cell ◽  
Small Interfering Rna ◽  
Glycogen Synthase ◽  
Glycogen Synthase Kinase 3Β ◽  
Phosphatidylinositol 3 Kinase ◽  
T Cell Factor ◽  
Ovarian Surface Epithelial ◽  
Cox 2 ◽  
Induced Apoptosis ◽  
Interfering Rna

Abstract Gonadotropins play a prominent role in ovarian function and pathology. We have shown that treatment with gonadotropins (FSH and LH/human chorionic gonadotropin) reduces the amount of N-cadherin with a concomitant induction of apoptosis in human ovarian surface epithelial (OSE) cells, but precise molecular mechanisms remain to be elucidated. Here, we demonstrated activation of β-catenin/T-cell factor (TCF) signaling by gonadotropins. We further showed that ectopic expression of N-cadherin was sufficient to recruit β-catenin to the plasma membrane, thereby blocking β-catenin/TCF-mediated transactivation in gonadotropin-treated cells. Transfection with β-catenin small interfering RNA or expression of dominant negative TCF inhibited apoptosis, whereas expression of dominant stable β-catenin (S37A) caused significant apoptosis, thus supporting a proapoptotic role for β-catenin/TCF in human OSE. In addition, we showed that gonadotropins enhanced β-catenin/TCF transcriptional activity through inactivation of glycogen synthase kinase-3β in a phosphatidylinositol 3-kinase/Akt-dependent manner, indicating cross talk between the phosphatidylinositol 3-kinase/Akt and β-catenin signaling pathways through glycogen synthase kinase-3β. Furthermore, gonadotropins increased cyclooxygenase-2 (COX-2) expression via the β-catenin/TCF pathway. COX-2 also played a role in gonadotropin-induced apoptosis, as treatment with the COX-2-specific inhibitor NS-398 or COX-2 small interfering RNA blocked gonadotropin-dependent apoptotic activity. These findings suggest that the participation of β-catenin in adhesion and signaling may represent a novel mechanism through which gonadotropins may regulate the cellular fate of human OSE.

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