Down-regulation Of Progesterone Receptor Research Articles

Blood Plasma Small Non-Coding RNAs as Diagnostic Molecules for the Progesterone-Receptor-Negative Phenotype of Serous Ovarian Tumors.

The expression level of the progesterone receptor (PGR) plays a crucial role in determining the biological characteristics of serous ovarian carcinoma. Low PGR expression is associated with chemoresistance and a poorer outcome. In this study, our objective was to explore the relationship between tumor progesterone receptor levels and RNA profiles (miRNAs, piwiRNAs, and mRNAs) to understand their biological characteristics and behavior. To achieve this, we employed next-generation sequencing of small non-coding RNAs, quantitative RT-PCR, and immunohistochemistry to analyze both FFPE and frozen tumor samples, as well as blood plasma from patients with benign cystadenoma (BSC), serous borderline tumor (SBT), low-grade serous ovarian carcinoma (LGSOC), and high-grade serous ovarian carcinoma (HGSOC). Our findings revealed significant upregulation of MMP7 and MUC16, along with downregulation of PGR, in LGSOC and HGSOC compared to BSC. We observed significant correlations of PGR expression levels in tumor tissue with the contents of miR-199a-5p, miR-214-3p, miR-424-3p, miR-424-5p, and miR-125b-5p, which potentially target MUC16, MMP7, and MMP9, as well as with the tissue content of miR-16-5p, miR-17-5p, miR-20a-5p, and miR-93-5p, which are associated with the epithelial-mesenchymal transition (EMT) of cells. The levels of EMT-associated miRNAs were significantly correlated with the content of hsa_piR_022437, hsa_piR_009295, hsa_piR_020813, hsa_piR_004307, and hsa_piR_019914 in tumor tissues. We developed two optimal logistic regression models using the quantitation of hsa_piR_020813, miR-16-5p, and hsa_piR_022437 or hsa_piR_004307, hsa_piR_019914, and miR-93-5p in the tumor tissue, which exhibited a significant ability to diagnose the PGR-negative tumor phenotype with 93% sensitivity. Of particular interest, the blood plasma levels of miR-16-5p and hsa_piR_022437 could be used to diagnose the PGR-negative tumor phenotype with 86% sensitivity even before surgery and chemotherapy. This knowledge can help in choosing the most effective treatment strategy for this aggressive type of ovarian cancer, such as neoadjuvant chemotherapy followed by cytoreduction in combination with hyperthermic intraperitoneal chemotherapy and targeted therapy, thus enhancing the treatment's effectiveness and the patient's longevity.

Bisphenol A Analogues Suppress Spheroid Attachment on Human Endometrial Epithelial Cells through Modulation of Steroid Hormone Receptors Signaling Pathway

Bisphenol A (BPA) is a well-known endocrine disruptor, widely used in various consumer products and ubiquitously found in air, water, food, dust, and sewage leachates. Recently, several countries have restricted the use of BPA and replaced them with bisphenol S (BPS) and bisphenol F (BPF), which have a similar chemical structure to BPA. Compared to BPA, both BPS and BPF have weaker estrogenic effects, but their effects on human reproductive function including endometrial receptivity and embryo implantation still remain largely unknown. We used an in vitro spheroid (blastocyst surrogate) co-culture assay to investigate the effects of BPA, BPS, and BPF on spheroid attachment on human endometrial epithelial cells, and further delineated their role on steroid hormone receptor expression. We also used transcriptomics to investigate the effects of BPA, BPS, and BPF on the transcriptome of human endometrial cells. We found that bisphenol treatment in human endometrial Ishikawa cells altered estrogen receptor alpha (ERα) signaling and upregulated progesterone receptors (PR). Bisphenols suppressed spheroid attachment onto Ishikawa cells, which was reversed by the downregulation of PR through PR siRNA. Overall, we found that bisphenol compounds can affect human endometrial epithelial cell receptivity through the modulation of steroid hormone receptor function leading to impaired embryo implantation.

Lost but Not Least-Novel Insights into Progesterone Receptor Loss in Estrogen Receptor-Positive Breast Cancer.

Simple SummaryMost breast cancers co-express estrogen receptor α (ERα) and progesterone receptor (PgR). These cancers are sensitive to endocrine therapy and, in general, have superior outcomes. However, a subset of tumors expresses ERα but loses expression of PgR in various mechanisms. The processes driving the loss of PgR may cause resistance to hormonal treatment and a more aggressive clinical course. The current review summarizes current knowledge on the biology of ERα-positive PgR(−)negative breast cancer and discusses the associations between molecular mechanisms and clinical characteristics.Estrogen receptor α (ERα) and progesterone receptor (PgR) are crucial prognostic and predictive biomarkers that are usually co-expressed in breast cancer (BC). However, 12–24% of BCs present ERα(+)/PgR(−) phenotype at immunohistochemical evaluation. In fact, BC may either show primary PgR(−) status (in chemonaïve tumor sample), lose PgR expression during neoadjuvant treatment, or acquire PgR(−) phenotype in local relapse or metastasis. The loss of PgR expression in ERα(+) breast cancer may signify resistance to endocrine therapy and poorer outcomes. On the other hand, ERα(+)/PgR(−) BCs may have a better response to neoadjuvant chemotherapy than double-positive tumors. Loss of PgR expression may be a result of pre-transcriptional alterations (copy number loss, mutation, epigenetic modifications), decreased transcription of the PGR gene (e.g., by microRNAs), and post-translational modifications (e.g., phosphorylation, sumoylation). Various processes involved in the down-regulation of PgR have distinct consequences on the biology of cancer cells. Occasionally, negative PgR status detected by immunohistochemical analysis is paradoxically associated with enhanced transcriptional activity of PgR that might be inhibited by antiprogestin treatment. Identification of the mechanism of PgR loss in each patient seems challenging, yet it may provide important information on the biology of the tumor and predict its responsiveness to the therapy.

P–289 Progesterone receptor is not downregulated in endometrial epithelial compartment during embryo receptivity phase in assisted reproductive cycles

Abstract Study question Is progesterone receptor (PGR) downregulation disrupted within endometrial epithelial compartment, during embryo receptivity phase in assisted reproductive technology (ART) cycles? Summary answer PGR is not downregulated in endometrial epithelial cells from ART cycles during embryo receptivity phase. What is known already Progesterone (P4) promotes the downregulation of its own progesterone receptor (PGR). During the mid-luteal phase, PGR is downregulated in endometrial epithelial cells (EEC), a critical process for embryo implantation. Embryos are unable to attach to the maternal surface when PGR expression is sustained in EEC. Non-physiologic ovarian steroid produced or employed in ART cycles may alter endometrial development compromising its receptivity. Scarce information is available whether PGR is downregulated in EEC from ARTs including ovarian stimulation for in vitro fertilization (IVF) cycles or hormonal endometrial preparation for frozen thawed embryo transfer (HEP-FET). Study design, size, duration Cross sectional study including endometrial samples from fertile women during natural cycle (FNC, n = 23), from infertile women submitted to IVF (n = 19) and from infertile women who underwent mock HEP-FET (n = 35). Samples were obtained between 2018–2019. Sample size was calculated considering a power of 90%, alpha error=0.05, an expected PGR expression of 2 and 0.5 in ART and FNC groups, respectively, having a standard deviation=0.9. At least 9 patients would be necessary in each group. Participants/materials, setting, methods Endometrial samples were obtained during mid-luteal phase scheduled 7 days after ovulation in FNC, 5 days after oocyte retrieval in IVF without embryo transfer or 5 days after P4 supplementation in HEP-FET. Immunohistochemistry was employed to quantify PGR using histologic score (Hscore). PGR mRNA levels were determined by qRT-PCR from EEC dissected by laser capture microdissection. Anova test was used for comparing means of Hscore and mRNA among groups. Statistical significance was established as P < 0.05. Main results and the role of chance No statistical differences were found in demographic characteristics including age, body mass index or endometrial thickness. The PGR expression was reduced in FNC compared to IVF and HP-FET endometria (0.6 ± 0.1, 1.9 ± 0.9 and 2.2 ± 0.9 respectively; P < 0.0001). The PGR mRNA levels from ECC dissected by laser capture microdissection were higher in IVF and HP-ET cycles compared to FNC (10.6 ± 3.1, 13.6 ± 2.3 and 0.8 ± 0.1 respectively; P < 0.0001) corroborating the elevated PGR Hscore in EEC from ART cycles. Limitations, reasons for caution This is a descriptive study reporting failure of PGR downregulation in endometria from ART cycles with vaginal P4 supplementation during the luteal-phase. Whether interference or resistance to P4 signal is the mechanism involved in the failure of PGR down regulation in ART cycles needs to be determined Wider implications of the findings: PGR downregulation within EEC was shown in FNC. The retained PGR expression detected in most ART cycles may interfere with embryo implantation and might explain the restricted pregnancy success. Future studies might reveal whether PGR evaluation in EEC can predict embryo implantation. Trial registration number Not Aplicable

Expression of nuclear progesterone receptor, progesterone receptor membrane components 1 and 2 and prostaglandin-endoperoxide synthase 2 in the endometrium and oviduct of spontaneously ovulating cats

Although ovulations not followed by pregnancy occur regularly in cats, differences in endometrial function between cats in the luteal and non-luteal phase have not been studied so far. Progesterone exerts its effects through a nuclear progesterone receptor (PGR) and via cell-membrane bound receptors referred to as progesterone receptor membrane component (PGRMC) 1 and 2. Progesterone receptor expression is regulated by gonadal steroid hormones and therefore may change throughout the oestrous cycle. Protein expression of PGR, PGRMC-1 and 2 and prostaglandin-endoperoxide synthase 2 (PTGS2) was analysed in the endometrium and oviduct of non-pregnant female cats in the follicular (n = 8) and luteal phase (n = 9). We hypothesized that the presence of corpora lutea (CL) is associated with downregulation of progesterone receptors and PTGS2. Cells of the luminal endometrial epithelium, endometrial stroma and oviductal epithelium were assessed by immunohistochemistry. The PGR protein expression was more pronounced in the endometrial epithelium than stroma (p < 0.001) and less pronounced in cats with a CL than without CL (p < 0.001) but did not differ between groups in the oviduct. The PTGS2 was localized only in the endometrial and oviductal epithelium and its expression was reduced in cats with CL (p = 0.001). In the endometrial epithelium, PGRMC-1 expression was reduced in cats with CL (p < 0.05). Expression of PGRMC-2 was highest in the endometrial epithelium and lowest in the endometrial stroma (p = 0.01) but did not differ between cats with and without CL. In conclusion, progesterone receptor and PTGS2 downregulation in the female cat closely resembles findings in other spontaneously ovulating domestic animal species.

Regulation of conceptus development and attachment in pigs

Implantation/placentation in domestic pigs is preceded by synthesis of oestrogen by the conceptus to maintain functional corpora lutea throughout pregnancy and a rapid morphological transformation of conceptuses from spherical to long filamentous thread-like structures. Initial conceptus expansion, reaching a metre in length, not only delineates the surface area for placental attachment, but also provides the mechanism for delivery of oestrogen to signal events necessary for placentation throughout the uterine horn. Timing for conceptus gene expression to induce trophoblast expansion and attachment in pigs is temporally associated with downregulation of progesterone receptors and increase in oestrogen receptors within the uterine epithelium. Within the confines of the uterine lumen, pig conceptuses normally do not erode or invade through the uterine epithelial surface. However, the pig conceptus possesses extensive proteolytic activity as it is highly invasive outside the uterine lumen of the pig. Initial release of oestrogen by the elongating pig conceptus induces endometrial release of cytokines and a variety of protease inhibitors. Recently, endometrial expression for the inter-trypsin inhibitor (I alpha I) family of protease inhibitors has been detected in the pig endometrium during conceptus elongation and attachment. It is possible that I alpha Is may function to inhibit trophoblast invasion and also serve as targets for adhesion molecules, such as integrins and heparin, to aid in placental attachment to the uterine epithelium.

51 Effects of modulating early luteal phase progestin concentration on endometrial function in early pregnant mares

Progesterone prepares the endometrium for pregnancy. This requires down-regulation of progesterone receptors in the endometrial epithelium as a prerequisite for the expression of pregnancy-associated proteins. We investigated effects of modulated peripheral progestin concentration in early luteal phase mares on endometrial function on Day 14 of pregnancy. Genitally healthy oestrous mares (n=8; age 4 to 14 years) were inseminated until ovulation and treated with either altrenogest (0.044mg kg−1 once daily orally) on Days 5 to 10 after ovulation (ALT), cloprostenol (125mg once daily intramuscularly) on Days 0 to 3 after ovulation (CLO) or left untreated (CON). The ALT and CLO treatments were chosen to increase and decrease total peripheral progestin concentration, respectively. Each treatment was given to every mare in consecutive cycles at random order. On Day 14 after ovulation, endometrial fluid was collected with a cotton roll (Salivette, Sarstedt, Germany) inserted into the uterus and an endometrial biopsy for immunohistological examination was collected. In endometrial fluid, free amino acid concentrations were analysed by ion exchange liquid chromatography with an amino acid analyser (Institut Kuhlmann, Analytik-Zentrum Ludwigshafen, Germany). Cell nuclei staining positive for the progesterone receptor were determined in the luminar and glandular epithelium as well as in the stroma. Statistical analysis was performed by non-parametric Friedman test with subsequent Wilcoxon test. Values are given as mean±standard error of mean. Pregnancy rate was 0.6±0.1 (13 cycles/8 pregnancies), 1.0±0 (8 cycles/8 pregnancies), and 0.7±0.1 (11 cycles/8 pregnancies) in CON, ALT, and CLO cycles, respectively (P=0.062). Conceptus size between Days 10 and 14 did not differ among treatments. The percentage of luminal epithelial cells staining positive for progesterone receptor differed among treatments (CON 72.8±4.1, ALT 70.7±4.7, and CLO 84.1±1.9%; P&amp;lt;0.05) and was higher in CLO than in ALT and CON cycles (P&amp;lt;0.05). Free amino acids glutamic acid and glycine were most abundant in endometrial fluid, but their concentrations did not differ among treatments. The concentrations of the amino acids isoleucine (CON 0.17±0.03, CLO 0.14±0.02, and ALT 0.23±0.04 µmol) and lysine (CON 0.27±0.08, CLO 0.18±0.05, and ALT 0.44±0.13 µmol) were influenced by treatment (P&amp;lt;0.05) and lower in CLO than in ALT and CON cycles. In conclusion, impaired luteal function due to CLO treatment during the early luteal phase of pregnant mares delayed down-regulation of progesterone receptors in the endometrial epithelium on Day 14. This influenced endometrial function as reflected in lower concentrations of the amino acids lysine and isoleucine in endometrial secretions.

Roles of SIRT1/FoxO1/SREBP-1 in the development of progestin resistance in endometrial cancer.

The prevalence of endometrial cancer (EC) is increasing worldwide. Progestin therapy is effective for both early stage EC patients who require preserving fertility and advanced or recurrent patients. Progestin resistance resulting from downregulation of progesterone receptor (PR) remains a major problem, and its mechanism is currently unclear. It was demonstrated that Sirtuin 1 (SIRT1), forkhead transcription factor 1 (FoxO1) and sterol regulatory element binding protein-1 (SREBP-1) may act as a pathway and play crucial roles in the development of EC in our previous studies. In the present study, we investigated the effect on the development of progestin resistance and the relationship with PR of SIRT1/FoxO1/SREBP-1. A progestin-resistant Ishikawa cell line was established in the stimulation and selection of medroxyprogesterone acetate (MPA), and the resistance was analyzed by MTT assay, flow cytometry, and Transwell invasion assay. qRT-PCR and western blotting were conducted to detect the expression of SIRT1, FoxO1, SREBP-1 and PR. SIRT1 knockdown progestin-resistant cells were established by lentiviral transduction. The new progestin-resistant cell line presented sufficient resistance to MPA in aspects of proliferation, distribution of cell cycle and apoptosis compared with original Ishikawa cells. Besides, the invasion capability of progestin-resistant cells was observably increased. In both protein and mRNA levels, SIRT1 and SREBP-1 were upregulated in progestin-resistant cells, while PR and FoxO1 were downregulated. SIRT1 was knocked down by lentivirus transfection in progestin-resistant cells, resulting in upregulation of PR, FoxO1 and downregulation of SREBP-1, thereby SIRT1 knockdown cells were more sensitive to MPA compared with progestin-resistant cells. SIRT1/FoxO1/SREBP-1 act as a pathway targeting PR and involve in the development of progestin resistance in Ishikawa cells.

Abstract 5418: GATA3 zinc-finger mutation induces transcriptional reprogramming in breast cancer through gain and loss of function

Abstract Recent large-scale breast cancer genomic profiling identified frequent mutations in a pioneer transcription factor, GATA3. The expression of GATA3 is a prominent marker of luminal breast tumors, and loss of GATA3 expression is associated with aggressive tumor phenotypes. In addition, GATA3 mutations are frequently found in these luminal tumors. However, the clinical and molecular outcomes of GATA3 mutations are poorly understood. Our novel classification scheme of GATA3 mutations defines distinct clinical features of breast tumors with mutations in the second GATA3 zinc-finger (ZnFn2): frequent observation in luminal B subtype and worse prognosis than other GATA3 mutant cases. To dissect the molecular impacts of these ZnFn2 mutations, we generated a GATA3 mutant breast cancer cell clone by CRISPR-Cas9 gene editing. An engineered ZnFn2 mutant cell line manifested increased tumor growth as xenografts and more aggressive phenotypes in vitro. The ZnFn2 mutation led to loss of GATA3 binding and decreased expression at a subset of genes, including progesterone receptor (PR). The mutant cells also exhibited gain of GATA3 binding at other loci, correlated with increased expression of mesenchymal marker genes. Decreased expression of PR and impaired response to progesterone were crucial for cancer-promoting functions in GATA3 ZnFn2 mutant cells. Downregulation of PR and its downstream genes was also observed in the clinical gene expression data. These results illuminate tumor-promoting functions of GATA3 ZnFn2 mutations in breast cancer. Citation Format: Motoki Takaku, Sara A. Grimm, John D. Roberts, Kaliopi Chrysovergis, Page Myers, Charles M. Perou, Paul A. Wade. GATA3 zinc-finger mutation induces transcriptional reprogramming in breast cancer through gain and loss of function [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5418.

MiR-129-2 mediates down-regulation of progesterone receptor in response to progesterone in breast cancer cells

ABSTRACTObjective: Hormonal therapy is an important component of first line of treatment for breast cancer. Response to hormonal therapy is influenced by the progesterone receptor (PR)-status of breast cancer patients. However as an early effect, exposure to progesterone decreases expression of PR in breast cancer cells. An understanding of the mechanism underlying down-regulation of PR could help improve response to hormonal therapy. Methods: We performed small RNA sequencing of breast cancer cells for identification of microRNAs targeting PR in response to progesterone treatment. Biochemical approaches were used to validate the findings in breast cancer cells. Results: Analysis of small RNA sequencing of four breast cancer cell lines treated with progesterone revealed an up-regulation of miR-129-2 independent of the PR status of the cells. We show that miR-129-2 targets 3′UTR of PR to down-regulate its expression. Furthermore, inhibition of miR-129-2 expression rescues the down-regulation of PR in breast cancer cells. Also, the expression levels of miR-129-2 was observed to be elevated in patients with low expression of PR in the TCGA cohort (n = 359). Conclusion: miR-129-2 mediates down-regulation of PR in breast cancer cells in response to progesterone, while anti-miR-129-2 could potentiate PR expression levels among patients with inadequate PR levels. Thus, modulation of activity of miR-129-2 could stabilize PR expression and potentially improve response to hormonal therapy under adjuvant or neo-adjuvant settings.

Abstract 2556: Loss of progesterone receptor through epigenetic regulation is associated with poor prognosis in multiple solid tumors

Abstract Introduction: As a tumor suppressor in the endometrium, progesterone and its synthesized analogue progestin have a long history as a treatment for endometrial cancer. Loss of progesterone receptor (PR) in endometrial cancer leads to therapeutic failure, and our group has identified several mechanisms underlying PR loss, most notably epigenetic silencing of PR transcription. Recently, a Finnish group demonstrated that progestin therapy in premenopausal women is associated with a lower incidence of not only endometrial cancer, but also ovarian, pancreatic and lung cancers. This unexpected protective function of progestin in organs outside of the reproductive system led us to hypothesize that tumor progression effects in endometrial, ovarian, pancreatic and lung cancer is occurs due to the loss of progesterone’s protective effects. Methods and results: Supporting a potential protective role of progesterone, we discovered that PR is downregulated in multiple tumors, including breast, endometrial, ovarian, cervical, pancreatic and lung cancers, and loss of PR was consistently associated with a poor prognosis based on analysis of the Oncomine database. Similarly, PR expression was low in multiple ovarian, pancreatic and lung cancer cells. Mechanistic studies revealed that PR downregulation was mediated by the polycomb-repressor complex and DNA methylation, though the precise mechanism differed among cell lines and cancer types. Treatment with epigenetic modulators, which are FDA-approved for multiple myeloma, restored functional PR expression at both the mRNA and protein level and promoted marked cell death through induction of apoptosis. . Moreover, overexpression of PR or treatment with epigenetic modulators in combination with progesterone enhanced this effect, indicating a key protective role for progesterone signaling through PR. Conclusion: These data suggest loss of PR may be a general theme for tumorigenesis or disease progression in multiple tumor types beyond the classically studied endometrial cancer. In addition, the epigenetic mechanisms contributing to PR downregulation in endometrial tumors were recapitulated in other tumor types, including ovarian, pancreatic and lung tumors. These studies collectively set the stage for use of progestin therapy in combination with epigenetic modulators, a concept we term “molecularly enhanced progestin therapy,” as a novel therapeutic approach for many cancers. This combinatorial strategy has the power to revitalize the use of epigenetic modulators, which have proved disappointing in solid tumors, as an approach to resensitize cancer cells to a tumor suppressor. Citation Format: Yiyang Li, Tamar Kavlashvili, Cheng Huang, Yuping Zhang, Xiangbing Meng, Kristina Thiel, Kimberly Leslie, Shujie Yang. Loss of progesterone receptor through epigenetic regulation is associated with poor prognosis in multiple solid tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2556. doi:10.1158/1538-7445.AM2017-2556

Fibroblast growth factor signalling induces loss of progesterone receptor in breast cancer cells.

We have recently demonstrated that, fibroblast growth factor 2 (FGFR2), signalling via ribosomal S6 kinase 2 (RSK2), promotes progression of breast cancer (BCa). Loss of progesterone receptor (PR), whose activity in BCa cells can be stimulated by growth factor receptors (GFRs), is associated with poor patient outcome. Here we showed that FGF7/FGFR2 triggered phosphorylation of PR at Ser294, PR ubiquitination and subsequent receptor`s degradation via the 26S proteasome pathway in BCa cells. We further demonstrated that RSK2 mediated FGF7/FGFR2-induced PR downregulation. In addition, a strong synergistic effect of FGF7 and progesterone (Pg), reflected in the enhanced anchorage-independent growth and cell migration, was observed. Analysis of clinical material demonstrated that expression of PR inversely correlated with activated RSK (RSK-P) (p = 0.016). Patients with RSK-P(+)/PR(–) tumours had 3.629-fold higher risk of recurrence (p = 0.002), when compared with the rest of the cohort. Moreover, RSK-P(+)/PR(–) phenotype was shown as an independent prognostic factor (p = 0.006). These results indicate that the FGF7/FGFR2-RSK2 axis promotes PR turnover and activity, which may sensitize BCa cells to stromal stimuli and contribute to the progression toward steroid hormone negative BCa.

Preterm delivery and associated negative pregnancy outcome – A tale of faulty progesterone receptor signalling pathway and linked derailed immunomodulation: A study from Northeast India

Preterm delivery (PTD) is one of the potent contributor of neonatal mortality and morbidity, and the underlying cause in some situation is elusive. This study attempts to delineate the association of deregulation in progesterone receptor (PR) pathway and deleterious immune responses in predisposing patients to PTD in Northeast India, a region with high rate of PTD cases. A total of 109 cases of PTD and 100 term delivery cases were enrolled with all clinical details. The PTD cases were stratified based on gestation age at delivery. The differential expression of PR and key downstream effectors and cytokines were evaluated for correlation with PTD susceptibility, gestational period, and pregnancy outcome. The results indicated a sharp downregulation in PR expression is associated with PTD susceptibility, lower gestational period and negative pregnancy outcome. The PR downstream effector PIBF was also found to be downregulated in PTD, and is associated with gestational period and negative pregnancy outcome. The downregulation of PR and PIBF expression was found to correlate with a predominant Th1 state with higher CD56+NK cell counts and pro-inflammatory burst lead by hyper TNF-α, NF-kB and IFNγ expression, and complicated by lower IL10 expression, contributing to PTD as well as negative pregnancy outcome in the PTD cases. TNF-α expression in placenta inversely correlated with placental PR expression. To conclude, deregulation in PR pathway is a hallmark of preterm delivery and negative pregnancy outcome. Differential expression of several markers such as PR, PIBF and TNF-α has prognostic significance, and hence is of clinical significance.

Clinical correlates of leiomyoma estrogen and progesterone receptors among Nigerian women

ObjectiveTo compare the expression of estrogen receptor α (ERα) and progesterone receptor (PR) in myometrium and leiomyomata tissue, and to correlate their expression with symptoms of uterine leiomyomata. MethodsIn a cross-sectional study, intraoperative biopsy samples of leiomyomata and adjacent myometrial specimens were obtained from premenopausal women with uterine leiomyomata treated at a center in Nigeria between September 2013 and August 2014. Immunohistochemistry for ERα and PR expression was performed on the samples. The immunoscores of both receptors were correlated with the size and symptoms of the leiomyomata. ResultsAmong 60 pairs of samples, leiomyomata had a higher mean expression of ERα (H-score 193.42±64.55 vs 153.29±69.13; P=0.01) and PR (214.86±66.56 vs 171.53±63.53; P<0.001) than did myometrial tissues. The tumor diameter correlated negatively with the immunoscores of both receptors irrespective of age, parity, and body mass index, but this was only significant for PR (ρ=–0.44; P<0.001). Downregulation of PR on leiomyomata was predicted to occur at a diameter of 11cm. Menorrhagia, dysmenorrhea, and infertility occurred independently of steroid-receptor expression. ConclusionLeiomyomata seem to depend on steroid hormones, but only during early tumor development. This could have implications for the selection of patients for medical management, especially with steroid-receptor modulators.

Chronic Exposure to Bisphenol A Affects Uterine Function During Early Pregnancy in Mice.

Environmental and occupational exposure to bisphenol A (BPA), a chemical widely used in polycarbonate plastics and epoxy resins, has received much attention in female reproductive health due to its widespread toxic effects. Although BPA has been linked to infertility and recurrent miscarriage in women, the impact of its exposure on uterine function during early pregnancy remains unclear. In this study, we addressed the effect of prolonged exposure to an environmental relevant dose of BPA on embryo implantation and establishment of pregnancy. Our studies revealed that treatment of mice with BPA led to improper endometrial epithelial and stromal functions thus affecting embryo implantation and establishment of pregnancy. Upon further analyses, we found that the expression of progesterone receptor (PGR) and its downstream target gene, HAND2 (heart and neural crest derivatives expressed 2), was markedly suppressed in BPA-exposed uterine tissues. Previous studies have shown that HAND2 controls embryo implantation by repressing fibroblast growth factor and the MAPK signaling pathways and inhibiting epithelial proliferation. Interestingly, we observed that down-regulation of PGR and HAND2 expression in uterine stroma upon BPA exposure was associated with enhanced activation of fibroblast growth factor and MAPK signaling in the epithelium, thus contributing to aberrant proliferation and lack of uterine receptivity. Further, the differentiation of endometrial stromal cells to decidual cells, an event critical for the establishment and maintenance of pregnancy, was severely compromised in response to BPA. In summary, our studies revealed that chronic exposure to BPA impairs PGR-HAND2 pathway and adversely affects implantation and the establishment of pregnancy.

Expression of estrogen receptor 1 and progesterone receptor in primary goat mammary epithelial cells.

The exact role and sensitivity of cells to estrogen and progesterone mediated through the steroid receptors during lactation is not known. Expression of estrogen receptor 1 (ESR1) and progesterone receptor (PGR) was quantified in mammary tissue-derived primary goat mammary epithelial cells (pgMECs) to determine the influence of donor tissue physiology (lactating and juvenile) and cell culture growth conditions (basal and lactogenic) on ESR1 and PGR expression in the derived cells. Relative messenger RNA (mRNA) levels for both receptors were the highest in cell lines derived from mammary tissue of juvenile goats. Maintaining pgMECs in lactogenic conditions resulted in up-regulation of ESR1 (1.36- to 12.35-fold) and in down-regulation of PGR (-2.53- to -3.62-fold), compared to basal conditions. Based on Western blotting analysis we suggest that the differences in mRNA expression are translated to the protein level. We suggest that differential expression in lactating conditions is correlated with terminal differentiation of the pgMECs. Double immunostainings showed that estrogen receptor alpha (ER-α) positive cells do not exclusively belong to the luminal lineage and that ER-α and PGR can be expressed individually or co-expressed in the pgMECs. The derived primary cultures/lines in early passages are hormone-responsive and represent a useful surrogate for mammary tissue in research experiments.

Chronic exposure to bisphenol a impairs progesterone receptor-mediated signaling in the uterus during early pregnancy.

Environmental and occupational exposure to endocrine disrupting chemicals (EDCs) is a major threat to female reproductive health. Bisphenol A (BPA), an environmental toxicant that is commonly found in polycarbonate plastics and epoxy resins, has received much attention due to its estrogenic activity and high risk of chronic exposure in human. Whereas BPA has been linked to infertility and recurrent miscarriage in women, the impact of its exposure on uterine function during early pregnancy remains unclear. In a recent publication in Endocrinology, we demonstrated that prolonged exposure to an environmental relevant dose of BPA disrupts progesterone receptor-regulated uterine functions, thus affecting uterine receptivity for embryo implantation and decidua morphogenesis, two critical events for establishment and maintenance of early pregnancy. In particular we reported a marked impairment of progesterone receptor (PGR) expression and its downstream effector HAND2 in the uterine stromal cells in response to chronic BPA exposure. In an earlier study we have shown that HAND2 controls embryo implantation by repressing fibroblast growth factor (FGF) expression and the MAP kinase signaling pathway, thus inhibiting epithelial proliferation. Interestingly we observed that downregulation of PGR and HAND2 expression in uterine stroma upon BPA exposure was associated with an enhanced activation of FGFR and MAPK signaling, aberrant proliferation, and lack of uterine receptivity in the epithelium. In addition, the proliferation and differentiation of endometrial stromal cells to decidual cells, an event critical for the maintenance of early pregnancy, was severely compromised in response to BPA. This research highlight will provide an overview of our findings and discuss the potential mechanisms by which chronic BPA impairs PGR-HAND2 pathway and adversely affects implantation and the establishment of pregnancy.

Understanding the regulation of pituitary progesterone receptor expression and phosphorylation.

Administration of human FSH (hFSH) during the diestrus phase in cyclic rats is followed by a reduction in the preovulatory LH surge. This inhibitory action of FSH involves a decrease in the stimulatory effect of gonadotrope progesterone receptor (PR) activation, in a ligand-dependent (progesterone) and -independent (GNRH) manner. PR activation and action are mandatory for LH surge, and are dependent on the phosphorylation of serine (Ser) residues. Together with this post-translational modification, PR is marked for downregulation by proteasome machinery. These experiments used the western blotting technique to measure pituitary expression of PR-A and PR-B isoforms and phosphorylation levels of Ser294 and Ser400 PR-B in rats bearing i) hFSH treatment or ii) PR downregulation. Treatment with hFSH reduced LH secretion and increased that of estradiol in proestrus afternoon. hFSH injections, without altering PR-A and PR-B content or ratio, caused a reduction in phosphorylation of Ser294 and Ser400 but only when pituitaries were previously challenged with progesterone or GNRH for 2 h. However, while pSer294 levels increased after 2 h of pituitary incubation with progesterone or GNRH, those of pSer400 were not modified by these in vitro treatments. Finally, progesterone had a biphasic effect: in 2-h incubations increased pituitary PR-A and PR-B content, but after 8 h caused downregulation and altered PR-A:PR-B ratio. The results provide a potential mechanism through which LH levels are decreased by hFSH administration and better understanding of the control of PR expression and phosphorylation in rat pituitaries.

MiR-888: A Novel Cancer-Testis Antigen that Targets the Progesterone Receptor in Endometrial Cancer

Cancer-testis (CT) antigens are a large family of genes that are selectively expressed in human testis germ cells, overexpressed in a variety of tumors and predominantly located on the X chromosome. To date, all known CT antigens are protein-coding genes. Here, we identify miR-888 as the first miRNA with features characteristic of a CT antigen. In a panel of 21 normal human tissues, miR-888 expression was high in testes and minimal or absent in all other examined tissues. In situ hybridization localized miR-888 expression specifically to the early stages of sperm development within the testes. Using The Cancer Genome Atlas database, we discovered that miR-888 was predominately expressed in endometrial tumors, with a significant association to high-grade tumors and increased percent invasion. In a separate panel of endometrial tumor specimens, we validated overexpression of miR-888 by real-time polymerase chain reaction. In addition, miR-888 expression was highest in endometrial carcinosarcoma, a rare and aggressive type of endometrial tumor. Moreover, we identified the progesterone receptor (PR), a potent endometrial tumor suppressor, as a direct target of miR-888. These data define miR-888 as the first miRNA CT antigen and a potential mediator of an aggressive endometrial tumor phenotype through down-regulation of PR.

Solitary and Multiple Meningiomas: an Immunohistochemical Comparison

Summary Introduction. Meningiomas are common primary tumors of brain meninges. These neoplasms develop from arachnoid cap cells and are multiple in 1 - 10% cases. Occasionally, significantly higher multiplicity rates have been reported, at least partially due to the increased application of computed tomography and magnetic resonance imaging in the diagnostics of intracranial pathologies. Meningiomas generally express progesterone receptors, but only few studies have focused on sex hormone receptor differences between solitary and multiple meningiomas. Similarly, there is limited information on cell proliferation and adhesion factors in solitary and multiple meningiomas. Aim of the study. Was to evaluate the immunohistochemical differences in sex hormone receptor expression, cell proliferation and adhesion within solitary and multiple meningiomas. Material and methods. In a retrospective study, 11 consecutive multiple meningiomas and 20 grade-matched solitary meningiomas were assessed by immunohistochemistry (IHC) to detect estrogen receptors (ER), progesterone receptors (PR), Ki-67 and neural cell adhesion molecule (NCAM). IHC was followed by quantitative microscopic evaluation. Descriptive and inferential statistics was applied including confidence interval (CI) analysis, Mann-Whitney U test and Spearmen correlation by IBM SPSS Statistics 22.0 software; p values less than 0.05 were regarded as statistically significant. Results. Although PR were found in all samples, the mean expression was significantly lower in multiple meningiomas (p = 0.03): 30.0% (95% CI: 10.4 - 49.8) versus 70.6% (95% CI: 56.6 - 84.7) in solitary meningiomas. ER were invariably absent in both groups. The proliferation index did not differ in solitary and multiple tumors. There was a trend (p = 0.07) to higher mean expression of NCAM in multiple meningiomas than in control group: 48.3% (95% CI: 25.8 - 70.8) versus 24.6% (95% CI: 13.2 - 36.0), respectively. The multiple meningiomas showed diverse histological types and immunophenotypes in up to 33.3% patients. Conclusions. Multiple meningiomas are characterized by significant down-regulation of PR and up-regulation of NCAM. The last finding can indicate neural differentiation and/ or peculiarities of cell adhesion and signaling that facilitate multifocal proliferation. Diverse histological types as well as PR and NCAM expression in separate meningiomas within same patient indicate independent multicentric origin.