Human progesterone receptor A and B isoforms in CHO cells. I. Stable transfection of receptor and receptor-responsive reporter genes: transcription modulation by (anti)progestagens

Dijkema, R.; Schoonen, W.G.E.J.; Teuwen, R; Struik, E van der; Ries, R.; Kar, Bibekananda; Olijve, Wiebe

doi:10.1016/s0960-0760(97)00160-x

Cited by 42 publications

(17 citation statements)

References 31 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…3A). The concentration of MPA necessary to fully inhibit Wnt/β-catenin signaling is ∼0.1 nmol/L, and this is close to the reported dissociation constant for both progesterone receptor isoforms (40). Using this ligand concentration (0.1 nmol/L), molecular excess of the antiprogestagen Org-31489 could fully reverse Wnt/β-catenin signaling inhibition.…”

Section: Resultssupporting

confidence: 72%

Progesterone Inhibition of Wnt/β-Catenin Signaling in Normal Endometrium and Endometrial Cancer

Wang

Hanifi-Moghaddam

Hanekamp

et al. 2009

Clinical Cancer Research

120

114

View full text Add to dashboard Cite

Purpose. Wnt signaling regulates the fine balance between stemness and differentiation. Here, the role of Wnt signaling to maintain the balance between estrogen-induced proliferation and progesterone-induced differentiation during the menstrual cycle, as well as during the induction of hyperplasia and carcinogenesis of the endometrium, was investigated. Experimental Design: Endometrial gene expression profiles from estradiol (E 2 ) and E 2 + medroxyprogesterone acetate-treated postmenopausal patients were combined with profiles obtained during the menstrual cycle (PubMed; GEO DataSets). Ishikawa cells were transfected with progesterone receptors and Wnt inhibitors dickkopf homologue 1 (DKK1) and forkhead box O1 (FOXO1), measuring Wnt activation. Expression of DKK1 and FOXO1 was inhibited by use of sequence-specific short hairpins. Furthermore, patient samples (hormone-treated endometria, hyperplasia, and endometrial cancer) were stained for Wnt activation using nuclear β-catenin and CD44. Results: In vivo, targets and components of the Wnt signaling pathway (among them DKK1 and FOXO1) are regulated by E 2 and progesterone. In Wnt-activated Ishikawa cells, progesterone inhibits Wnt signaling by induction of DKK1 and FOXO1. Furthermore, using siRNA-mediated knockdown of both DKK1 and FOXO1, progesterone inhibition of Wnt signaling was partly circumvented. Subsequently, immunohistochemical analysis of the Wnt target gene CD44 showed that progesterone acted as an inhibitor of Wnt signaling in hyperplasia and in well-differentiated endometrial cancer. The female sex hormones estradiol (E 2 ) and progesterone play rate-limiting roles in the cyclical renewal of the inner layer of the uterus (endometrium). In the first half of the regular menstrual cycle, the proliferation phase, E 2 is required to expand the endometrial layer by inducing cell proliferation. In the second half of the menstrual cycle, the secretory phase, progesterone levels increase, which antagonizes the proliferative activity of E 2 by inducing differentiation of epithelial and stromal cells of the endometrium (1). Thus, inhibition of E 2 -induced proliferation by progesterone is crucial for the maintenance of homeostasis in the endometrium.Increased estrogen signaling often underlies endometrial hyperplasia and is a well-established risk factor for endometrial cancer (2). Because progesterone inhibits estrogen-induced endometrial proliferation, progesterone has been used in its synthetic form [i.e., medroxyprogesterone acetate (MPA)] in palliative treatment of advanced and recurrent endometrial cancer with modest though significant response rates (15-25%; ref. 3). Progesterone has also been used as a primary treatment for endometrial carcinoma confined to the endometrial layer of the uterus, for example, in premenopausal women determined to preserve fertility. Response rates in these women can be up to 60% (4, 5), indicating that progesterone signaling in well-differentiated endometrial cancer is a potent inhibitor of endometrial carcinogene...

show abstract

Section: Resultssupporting

confidence: 72%

Progesterone Inhibition of Wnt/β-Catenin Signaling in Normal Endometrium and Endometrial Cancer

Wang

Hanifi-Moghaddam

Hanekamp

et al. 2009

Clinical Cancer Research

120

114

View full text Add to dashboard Cite

show abstract

“…This is also supported by our functional data that P4 decreased HERG K ϩ current density in CHO cells (supplemental Fig. 2), which do not express P4 receptors (23). Cycloheximide, a protein synthesis inhibitor significantly decreased the total amount of HERG protein but failed to prevent/abolish the effect of P4 on HERG K ϩ channel maturation (Fig.…”

Section: Effect Of P4 Is Neither P4 Receptor-mediated Nor Via De Novosupporting

confidence: 81%

“…We found that RU486, a P4 receptor blocker, failed to reverse the effect of P4 on HERG K ϩ channel trafficking. In addition, the inhibitory effect of P4 on HERG currents in CHO cells, a P4 receptornegative cell line (23), further supports the hypothesis that the effect of P4 is nuclear receptor-independent. In addition, P4 did not decrease the total amount of HERG protein.…”

Section: Discussionsupporting

confidence: 68%

Progesterone Impairs Human Ether-a-go-go-related Gene (HERG) Trafficking by Disruption of Intracellular Cholesterol Homeostasis

Soong

et al. 2011

Journal of Biological Chemistry

View full text Add to dashboard Cite

The prolongation of QT intervals in both mothers and fetuses during the later period of pregnancy implies that higher levels of progesterone may regulate the function of the human ether-ago-go-related gene (HERG) potassium channel, a key ion channel responsible for controlling the length of QT intervals. Here, we studied the effect of progesterone on the expression, trafficking, and function of HERG channels and the underlying mechanism. Treatment with progesterone for 24 h decreased the abundance of the fully glycosylated form of the HERG channel in rat neonatal cardiac myocytes and HERG-HEK293 cells, a cell line stably expressing HERG channels. Progesterone also concentration-dependently decreased HERG current density, but had no effect on voltage-gated L-type Ca Progesterone (P4) 2 is an important steroid hormone involved in the female menstrual cycle, pregnancy, and embryogenesis. In the normal menstrual cycle, P4 levels increase from 0.6 -4.5 nmol/liter during the preovulatory phase to 10.5-80 nmol/liter during the luteal phase (1). It was reported that P4 at Ͻ100 nmol/liter, which is comparable with that occurring during the luteal phase, had no significant effect on the QT interval (2-4). However, recent studies show that P4 may shorten the action potential duration or QT interval (5). This is probably because P4 at this level enhances the rapid component of the delayed rectifier K ϩ current and inhibits L-type Ca 2ϩ currents (6). If pregnancy occurs, P4 levels are initially maintained at the luteal level, but may increase to 1 mol/liter at term (7). Elevated levels of P4 are important for the implantation of the embryo and the maintenance of a conducive environment for the embryo. At such a high level, the corrected QT interval is significantly prolonged (8, 9). This explains why pregnant women are more susceptible to ventricular arrhythmias during pregnancy, labor, and delivery (10 -12). However, it is interesting to note that in the patients with inherited long QT syndrome (LQTS) the risk for cardiac events is not higher during pregnancy than during other periods in life (10, 13). These findings suggest that the mechanisms for pregnancy-induced cardiac events in normal healthy women are different from those in the patients with inherited LQTS.In the fetus, the P4 level was reported to be very high (ϳ4.5 mol/liter) in umbilical cord vein during late stages of pregnancy (7). At this stage, longer QT and corrected QT intervals were reported in fetal magnetocardiography and noninvasive fetal electrocardiography (14, 15). More importantly, the rate of sudden death is higher in the uterus than at most other times in the human life cycle. Although the major reasons remain unknown, several studies of newborns suggest that QT prolongation and the increased risk of ventricular arrhythmias may account for the significant mortality (16,17). This may be the same for stillbirth as several case reports indicate that LQTS is one cause for otherwise unexplained fetal demise (18).The human ether-a-go-go-related gene (HE...

show abstract

“…33 and 34 and Table 1). For example, removal of carbon 19 (substitution of a methyl group for hydrogen: norprogesterone, also see R5020 and Organon 2058) decreases binding affinity to mPR␣ (Table 1) but modestly increases it for the nPR (33,34). However, removal of the side chain and addition of a hydroxyl at C17 (i.e.…”

Section: Discussionmentioning

confidence: 99%

“…However, removal of the side chain and addition of a hydroxyl at C17 (i.e. testosterone, dihydrotestosterone, nandrolone), which practically eliminated binding to the human nPR, only reduced binding with mPR␣ to 22-7% that of P4 (33,34). Finally, the hu-mPR␣ also does not recognize C19 steroids (androgens) with a substitution of a hydrogen on C17 with an ethinyl group (i.e.…”

Section: Discussionmentioning

confidence: 99%

Steroid and G Protein Binding Characteristics of the Seatrout and Human Progestin Membrane Receptor α Subtypes and Their Evolutionary Origins

Thomas¹,

Pang²,

Dong³

et al. 2007

Endocrinology

267

260

View full text Add to dashboard Cite

A novel progestin receptor (mPR) with seven-transmembrane domains was recently discovered in spotted seatrout and homologous genes were identified in other vertebrates. We show that cDNAs for the mPR ␣ subtypes from spotted seatrout A LTHOUGH THE IMPORTANCE of rapid (i.e. nonclassical) steroid actions initiated at the cell surface through binding to steroid membrane receptors has become more widely accepted within the past few years, details of the initial steroid-mediated events, including the identities of the steroid membrane receptors and their mechanisms of action, remain unclear and are surrounded by controversy (1-3). There is clear evidence that a variety of receptor proteins are involved in initiating these nonclassical steroid actions in different cell models, including nuclear steroid receptors or nuclear steroid receptor-like forms (1, 2, 4), receptors for other ligands that also bind steroids (2, 5), and unidentified receptors with different characteristics from those of any known receptors (2, 6). Recently, a novel cDNA was discovered in spotted seatrout ovaries that has several characteristics of the progestin membrane receptor (mPR) mediating progestin induction of oocyte maturation in this species by a nongenomic mechanism (7). The seatrout cDNA (st-mPR␣) encodes a 40 kDa protein, which has seven transmembrane domains, and receptor activation alters pertussis toxin-sensitive adenylyl cyclase activity, both of which suggest stmPR␣ is a G protein-coupled receptor (GPCR) or GPCR-like protein (7). More than 20 closely related genes have been cloned from other vertebrate species, including three mPR subtypes in humans, named ␣, ␤, and ␥, which show high levels of expression in human reproductive, brain, and kidney tissues, respectively (8). The identification of a new class of putative steroid receptors, unrelated to nuclear steroid receptors, but instead related to GPCRs, provides a plausible explanation of how steroids can initiate rapid hormonal responses in target cells by activating receptors on the cell surface. There has been broad recognition of the potential significance of these findings (1, 9, 10) and also an extensive research effort to determine the distribution, hormonal regulation, and biological roles of the mPRs in various vertebrate models (11-16). However, critical information is still lacking on several key features of mPRs essential for clearly establishing this proposed alternative model of steroid action and for understanding its likely evolutionary origins.The st-mPR␣ protein has been localized to the plasma membrane of seatrout oocytes (7), but progestin binding and activation of signal transduction pathways in the plasma membranes of cells transfected with the st-mPR␣ and human mPRs remain to be demonstrated. To date, progestin binding has only First Published Online November 9, 2006Abbreviations: GPCR, G protein-coupled receptor; HLY3, hemolysin 3; hu-mPR␣, human membrane progestin receptor ␣; MMD, monocyte to macrophage differentiation protein; mPR, membrane progestin rece...

show abstract

Human progesterone receptor A and B isoforms in CHO cells. I. Stable transfection of receptor and receptor-responsive reporter genes: transcription modulation by (anti)progestagens

Cited by 42 publications

References 31 publications

Progesterone Inhibition of Wnt/β-Catenin Signaling in Normal Endometrium and Endometrial Cancer

Progesterone Inhibition of Wnt/β-Catenin Signaling in Normal Endometrium and Endometrial Cancer

Progesterone Impairs Human Ether-a-go-go-related Gene (HERG) Trafficking by Disruption of Intracellular Cholesterol Homeostasis

Steroid and G Protein Binding Characteristics of the Seatrout and Human Progestin Membrane Receptor α Subtypes and Their Evolutionary Origins

Contact Info

Product

Resources

About