Expression Patterns Of Estrogen Receptor Research Articles

Changes in expression of breast cancer tumor biomarkers between primary tumors and corresponding metastatic sites: common patterns and relationships with survival.

In metastatic breast cancer, differences in expression patterns of estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor-2 (HER2) between the primary tumor (PT) and metastatic site (MET) have been reported. However, there is limited understanding of the relationship of tumor subtype discordance and overall survival (OS). We evaluated patterns of ER/PR/HER2 in PTs and corresponding METs and assessed the relationship between these patterns and OS. Patients diagnosed at our center with metastatic breast cancer (2011-2020) were included. ER/PR were stratified as < 1%/1-10%/ > 10% by immunohistochemistry and HER2 as positive/negative by immunohistochemistry/FISH. Tumor subtypes were classified as ER or PR + /HER2-, HER2+ , or triple-negative. Biomarker discordance data from PTs to METs were analyzed for expression patterns. OS was assessed. Of 254 patients, 41 (16.1%) had synchronous and 213 (83.9%) had metachronous METs. Category change of ER/PR/HER2 expression was observed in 56 (22.0%), 117 (40.5%), and 30 (11.8%) patients, respectively. Tumor subtype changed in 56 (22.0%) patients. We identified a difference between PT and MET from ER > 10% to ER < 1% (n = 28,16.2% p < 0.01); PR > 10% to PR < 1% (n = 54,48.2%, p < 0.001); PR > 10% to PR 1-10% (n = 18,16.1%, p < 0.001), and ER or PR+/HER2-to triple-negative (n = 19,13.0%, p = 0.03). In log-rank analysis, change from an ER or PR+/HER2-(5-year OS 88.6%) PT to a HER2+(67.5%) or triple-negative (54.6%) MET was associated with decreased survival (p < 0.01); however, in multivariate analysis, discordant biomarker expression was not associated with decreased survival (p > 0.05). Tumor expression of ER/PR/HER2 can differ between the PT and MET. Loss of ER/PR expression is common and may be related to worse survival. Routine assessment of MET tumor markers could inform prognosis and therapeutic decision-making.

Abstract 2164: Patterns of estrogen receptor expression across 18,500 tumor samples obtained from 149 cancer types

Abstract Estrogen receptor alpha (ER) is a nuclear transcription factor with a pivotal role in the function of the reproductive organs and of other organ systems. It constitutes a well-established therapeutic target for various small molecules. ER immunohistochemistry is used in surgical pathology for the distinction of tumors of the breast and gynecological organs. To comprehensively determine ER expression in normal and neoplastic tissues, a tissue microarray containing 18,560 samples from 149 different tumor types and subtypes as well as 608 samples of 76 different normal tissue types was analyzed by immunohistochemistry. ER positivity was found in 55 of 149 tumor categories, and 27 of these tumor categories contained at least one case with strong ER staining. ER positivity was most seen in various subtypes of breast cancer (78.9-95.7%) and in several different tumor categories of the ovary and the endometrium (8.5-75.9%). Other commonly ER positive tumor entities included leiomyoma (65.9%), neuroendocrine tumor of the appendix (43.8%), leiomyosarcoma (22.9%), Sertoli Leydig cell tumor of the ovary (1 of 3 positive), rhabdomyosarcoma (16.7%), angiomyolipoma (10%), colorectal neuroendocrine carcinoma (9.1%), myoepithelial carcinoma of the salivary gland (8.3%), granulosa cell tumor of the ovary (8.3%), epithelial-myoepithelial carcinoma of the salivary gland (7.7%), squamous cell carcinomas of various sites of origin (0.8-6.7%), testicular teratoma (5.3%), Brenner tumor of the ovary (5.1%), Ewing sarcoma (5%), adenocarcinoma NOS of the salivary gland (4.8%), papillary thyroid carcinoma (3.3%), sarcoma, not otherwise specified (NOS) (2.8%), paraganglioma (2.6%), non-invasive papillary urothelial carcinoma, pTa G3 (2.4%), pulmonary adenocarcinoma (2.1%), neuroendocrine tumor of the ileum (2.1%), and adenocarcinoma of the prostate, Gleason 5+5 (1.2%). Among 1,411 evaluable invasive breast carcinomas of no special type (NST), absent or low ER immunostaining was significantly linked to high BRE grade (p&amp;lt;0.0001), advanced pT stage (p&amp;lt;0.0001), distant metastasis (p=0.0012), HER2 overexpression and reduced progesterone receptor expression (p&amp;lt;0.0001 each) but not to nodal metastasis and patient survival. Among 378 high grade serous ovarian cancer, low ER immunostaining was linked to nodal metastasis (p=0.0112). Our data provide a comprehensive overview on the pattern of ER expression in cancer. The data show that ER expression predominated in gynecological and breast tumors. However, occasional (strong) ER expression can also occur in a broad variety of non-breast and non-gynecological neoplasms. Citation Format: Florian Viehweger, David Dum, Anne Menz, Ria Uhlig, Andrea Hinsch, Doris Hoeflmayer, Christoph Fraune, Christian Bernreuther, Patrick Lebok, Soeren Weidemann, Guido Sauter, Maximilian Lennartz, Frank Jacobsen, Till Sebastian Clauditz, Till Krech, Andreas H Marx, Ronald Simon, Stefan Steurer, Sarah Minner, Eike Burandt, Natalia Gorbokon. Patterns of estrogen receptor expression across 18,500 tumor samples obtained from 149 cancer types [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2164.

Evaluation of estrogen receptor, progesterone receptor, and Ki 67 immunoexpression in epithelial ovarian tumors with histopathological correlation

Abstract Background: Ovarian cancer ranks sixth among women’s cancers globally and seventh among cancer-related deaths. Around 90% of ovarian cancers have an epithelial genesis. Steroid hormones like progesterone and estrogen largely influence the development of ovarian cancers. In ovarian neoplasms, the proliferation marker Ki 67 helps forecast the course of the disease. Objective: To study the immunohistochemical expression pattern of estrogen receptor (ER), progesterone receptor (PR), and Ki 67 in the surface epithelial ovarian tumors (SEOTs) and correlate the findings with the standard histopathological. Materials and Methods: This prospective study was conducted in the Department of Pathology, MKCG Medical College and Hospital, Brahmapur, from November 2020 to October 2022 on 62 surface epithelial ovarian tumor cases. All the patients were evaluated for standard histopathological parameters and immuno-histochemistry (IHC) status using ER, PR, and Ki 67 as primary antibodies. Results: A total of 62 cases of SEOTs constituted the study group, of which 42 (67.74%) were benign, 5 (8.06%) were borderline, and 15 (24.2%) were malignant on histopathological evaluation: serous SEOTs (41, 66.12%), mucinous SEOTs (18, 29.03%), endometrioid SEOTs (2, 3.22%), and clear cell carcinoma (1, 1.61%). ER expression was more in malignant tumors (10/15; 66.66%) than in borderline (3/5; 60%) and benign (11/42; 26.19%), but the expression of PR was more in benign (27/42; 64.28%), followed by borderline (3/5; 60%) and malignant (8/15; 53.33%) SEOTs. Expression of Ki 67 was found to be more malignant (14/15; 93.33%) than borderline (4/5; 80%) and benign (2/42; 4.76%) SEOTs. Conclusion: ER and Ki 67 were expressed more in malignant, followed by borderline and benign, whereas PR expression was more innocent than borderline and malignant.

Poor Endometrial Proliferation After Clomiphene is Associated With Altered Estrogen Action.

Suboptimal endometrial thickening is associated with lower pregnancy rates and occurs in some infertile women treated with clomiphene. To examine cellular and molecular differences in the endometrium of women with suboptimal vs optimal endometrial thickening following clomiphene. Translational prospective cohort study from 2018 to 2020 at a university-affiliated clinic. Reproductive age women with unexplained infertility treated with 100 mg of clomiphene on cycle days 3 to 7 who developed optimal (≥8mm; n = 6, controls) or suboptimal (<6mm; n = 7, subjects) endometrial thickness underwent preovulatory blood and endometrial sampling. The main outcome measures were endometrial tissue architecture, abundance and location of specific proteins, RNA expression, and estrogen receptor (ER) α binding. The endometrium of suboptimal subjects compared with optimal controls was characterized by a reduced volume of glandular epithelium (16% vs 24%, P = .01), decreased immunostaining of markers of proliferation (PCNA, ki67) and angiogenesis (PECAM-1), increased immunostaining of pan-leukocyte marker CD45 and ERβ, but decreased ERα immunostaining (all P < .05). RNA-seq identified 398 differentially expressed genes between groups. Pathway analysis of differentially expressed genes indicated reduced proliferation (Z-score = -2.2, P < .01), decreased angiogenesis (Z-score = -2.87, P < .001), increased inflammation (Z-score = +2.2, P < .01), and ERβ activation (Z-score = +1.6, P < .001) in suboptimal subjects. ChIP-seq identified 6 genes bound by ERα that were differentially expressed between groups (P < .01), some of which may play a role in implantation. Women with suboptimal endometrial thickness after clomiphene exhibit aberrant ER expression patterns, architectural changes, and altered gene and protein expression suggesting reduced proliferation and angiogenesis in the setting of increased inflammation.

Donor variation and sex hormone receptors in periodontal ligament cells

ObjectiveThis study evaluated the gene expression and protein synthesis of sex hormone receptors in human periodontal ligament cells (PDLCs) in relation to donor- and tooth-specific factors with the aim to clarify the debate about sex hormone receptors in PDLCs. DesignThe expression patterns of estrogen receptors (genes: ESR1 and ESR2; proteins: ERα and ERβ), androgen receptor (AR) and progesterone receptor (PR) were investigated in the context of immortalization status, previous orthodontic tooth movement (OTM), donor age, sex and hormonal stimulation in PDLCs from 14 healthy donors (male: n = 8, female: n = 6; adolescents: n = 8, adults: n = 6) using quantitative real-time polymerase chain reaction, Western blot and immunocytochemistry. ResultsFor ERβ, the full-length isoform ERβ1 and truncated variants were detected. For ERα, the expected isoform ERα66 was not observed, but a novel isoform ERα36 was detected. Immortalization status, previous OTM and donor age had no impact on ESR1 and ESR2 expression. Estradiol stimulation for 24 h doubled the ratio of ESR2/ESR1 in PDLCs from female but not male donors, indicating sex-specific patterns of receptor expression. AR and PR demonstrated insufficient protein synthesis in PDLCs. ConclusionsThe data revealed a pivotal role for and complex interplay between ERα and ERβ in human PDLCs regardless of variable donor characteristics. Therefore, PDLC biology might be altered in patients of each age group and both sexes due to hormonal changes. This should be kept in mind during periodontic and orthodontic treatment of patients with special hormonal status.

Assessment of breast cancer immunohistochemistry and tumor characteristics in Nigeria.

BACKGROUNDFemale breast cancer is the leading type of cancer worldwide with an incidence of approximately 2.1 million in 2018. Hormone receptor status plays a vital role in its management.AIMTo determine the molecular expression pattern of biomarkers in breast cancer and their correlation with tumor variables.METHODSThis prospective study was designed to analyze expression patterns of estrogen receptor(ER), progesterone receptor (PR) and human epidermal growth factor receptor(HER2/neu) in breast cancer patients. The dataset has been taken from the Department of Radiotherapy and Oncology of Usmanu Danfodiyo University Teaching Hospital Sokoto, Nigeria from 1 January 2015 to 2 December 2019. The dataset had 259 records and 7 attributes. SPSS version 23.0 for statistical analysis was used. The data analyzed demographic and other clinicopathological characteristics as categorical variables. The mean and standard deviation were determined for the quantitative variable.RESULTSA total of 259 breast cancer cases were included in the study. The mean age was 48.3 ± 11.0, with an age range of 26-80 years and a median age of 46 years. The morphological categories were invasive ductal carcinoma 258 (99.6%) and invasive lobular carcinoma 1 (0.4%). ER, positivity increased in 73 patients (50%) under the age of 50 years, as well as PR positivity increased in 34 patients (23.6%) under the age of 50 years. HER/2neupositivity decreased in 8 patients (5.6%) under the age of 50 years. Hormonal receptors were statistically significant with clinicopathological characteristics (P < 0.05).CONCLUSIONOur study showed that ER, PR and HER2/neuexpression had a strong correlation with age, tumor grade, tumor size and lymph node status. Hence, hormone receptor assessment is highly recommended because of its significance in clinical management and prognostication.

Optimization of LC‐ESI‐MS Quantification of Ventromedial Hypothalamic Nucleus Estradiol Content by Central Composite Design

Brain structures exhibit characteristic patterns of estrogen receptor expression and regulatory functions and local capacity to generate neuroestradiol, necessitating quantification of estradiol content of individual loci. Estradiol acts on ventromedial hypothalamic nucleus (VMN) substrates to control behavior and glucostasis. Current research utilized Analytical Quality of Design to optimize uHPLC‐electrospray ionization‐mass spectrometry (LC‐ESI‐MS) sensitivity for analysis of estradiol in VMN samples obtained by high‐resolution microdissection techniques. A Design of Experiments matrix was developed to assess effects of critical independent variables on estradiol chromatogram area. A second‐order polynomial equation was generated by statistical response surface methodology using central composite design to identify synergistic versus antagonistic factors and to predict singular/combinatory effects on response area. Estradiol, estrone, and estriol were used as optimizing compounds; ethynyl estradiol served as internal standard. Lysates of VMN tissue harvested by micropunch dissection were transferred to methanol‐water conditioned solid phase extraction syringe cartridges and treated with 2% formic acid. Estradiol content was extracted with methanol, then vacuum concentrated. Samples were treated with 2‐fluoro‐1‐methylpyridinium‐p‐toluenesulfonate (FMP‐TS) prior to application to a C18 column [0.2ml/min mobile phase (25:75, v/v) flow rate; 0.1% formic acid, acetonitrile], followed by spectrometric analysis of estradiol and internal standard responses at m/z 364.2 and 388.2, respectively. Optimized mass spectrometric vaporizer (320°C) and ion transfer tube (266 °C) temperatures and sheath (22 psig), auxiliary (5 psig), and sweep (0.7 psig) gas pressures resulted in maximum tissue estradiol response. Outcomes validate a novel combinatory methodology for estradiol quantification in precisely‐defined brain components.Support or Funding InformationNational Institutes of Health DK‐109382

Cyanidin-3-o-Glucoside Pharmacologically Inhibits Tumorigenesis via Estrogen Receptor β in Melanoma Mice.

Expression patterns of estrogen receptors [ERα, ERβ, and G-protein associated ER (GPER)] in melanoma and skin may suggest their differential roles in carcinogenesis. Phytoestrogenic compound cyanidin-3-o-glucoside (C3G) has been shown to inhibit the growth and metastatic potential of melanoma, although the underlying molecular mechanism remains unclear. The aim of this study was to clarify the mechanism of action of C3G in melanoma in vitro and in vivo, as well as to characterize the functional expressions of ERs in melanoma. In normal skin or melanoma (n = 20/each), no ERα protein was detectable, whereas expression of ERβ was high in skin but weak focal or negative in melanoma; and finally high expression of GPER in all skin vs. 50% melanoma tissues (10/20) was found. These results correspond with our analysis of the melanoma survival rates (SRs) from Human Protein Atlas and The Cancer Genome Atlas GDC (362 patients), where low ERβ expression in melanoma correlate with a poor relapse-free survival, and no correlations were observed between SRs and ERα or GPER expression in melanoma. Furthermore, we demonstrated that C3G treatment arrested the cell cycle at the G2/M phase by targeting cyclin B1 (CCNB1) and promoted apoptosis via ERβ in both mouse and human melanoma cell lines, and inhibited melanoma cell growth in vivo. Our study suggested that C3G elicits an agonistic effect toward ERβ signaling enhancement, which may serve as a potential novel therapeutic and preventive approach for melanoma.

Sex- and Age-Related Estrogen Signaling Alteration in Inflammatory Bowel Diseases: Modulatory Role of Estrogen Receptors.

The pathogenesis of inflammatory bowel diseases (IBD) seems to be associated with alterations of immunoregulation. Several lines of evidence suggest that estrogens play a role in the modulation of immune responses and may be related to the etiology of IBD. The purpose of this work was to examine the involvement of G protein-coupled estrogen receptor (GPER), estrogen receptor α (ERα), estrogen receptor β (ERβ) and ERα spliced variants ERα36 and ERα46 in Crohn’s disease (CD) and ulcerative colitis (UC). The studied group included 73 patients with IBD and 31 sex and age-related controls. No differences in serum levels of 17β-estradiol nor of CYP1A1 and SULT1E1 enzymes involved in estrogen catabolism were stated. The expression pattern of estrogen receptors in tissue samples was quantified using real-time PCR and Western blotting. Statistically significant up-regulation of GPER and ERα in both CD and UC as well as down-regulation of ERβ in CD patients was found. However, differences in the expression of estrogen receptors in CD and UC have been identified, depending on the sex and age of patients. In men, up-regulation of GPER, ERα and ERα46 expression was shown in CD and UC patients. In women under 50 years of age, GPER protein level increased in UC whereas ERβ expression tended to decrease in CD and UC patients. In turn, in women over 50 the protein level of ERα increased in UC while ERβ expression decreased in CD patients. Dysregulation of estrogen receptors in the intestinal mucosa of patients with CD and UC indicates that estrogen signaling may play a role in the local immune response and maintain epithelial homeostasis in a gender- and age-dependent manner.

ERβ modulates genistein's cisplatin-enhancing activities in breast cancer MDA-MB-231 cells via P53-independent pathway.

As one of the typical food-derived phytoestrogens, genistein (GEN) could bind to estrogen receptor (ER) and was reported to be closely related to breast cancer. Our former research showed that GEN interfered with the anti-tumor effects of cisplatin (CIS) in breast cancer MCF-7 (ERα+/ERβ-) cells. However, it is not clear whether ER expression pattern affects GEN's modulation on CIS's activity. In the present study, breast cancer ERβ knockdown (ERβKD) MDA-MB-231 (ERα-/ERβ+) cell model was established via ERβ RNAi lentivirus infection. The role of ERβ expression in GEN's bioeffects on cells' response to CIS was investigated and was further double-checked by pathway-specific inhibitor PHTPP. Consistent results were harvested through cell viability analysis, cell cycle distribution flow cytometry, TUNEL staining, and expression detection of key biomarkers, Bax, Bcl-2, P21, P53, and cleaved caspase-3. Compared with the control group, PHTPP-treated or ERβKD cells exhibited higher sensitivity to both GEN and CIS treatment. GEN and CIS showed synergistic effects only in ERβ-deficient cells. This effect mainly resulted in G2 phase arresting and apoptosis induction with the upregulation of P21 and Bax/Bcl-2 protein level. Besides, P53 expression was strikingly suppressed in ERβ-deficient cells. This indicated ERβ pathway deficiency might enhance GEN-CIS bioactivity via the downregulation of P53. In summary, our data imply that daily intake of GEN-rich diet could collaborate with CIS anti-tumor treatment in ERα-/ERβ- breast cancer cases. ERβ pathway might be one of the potential targets which elicit GEN's positive effects in ERα- breast cancer patients.

Expression pattern of estrogen receptor β and its correlation with multidrug resistance in non-small cell lung cancer.

The aim of this work was to determine the expression of the ERβ (estrogen receptor β) and multidrug resistance, namely MDR1 (P-glycoprotein, P-gp), in 152 samples of non-small cell lung cancer. The expression pattern of ERβ and MDR1 were assessed by the quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) and immunohistochemistry. We also analyzed the correlation between ERβ and MDR1 with clinical and pathological data. The co-expression pattern of ERβ and individual MDR1 proteins was assessed by correspondence analysis and chi-squared tests. In the present study, we found that patients with tumor stage I-II showed higher ERβ mRNA expression levels and decreased expression of ERβ protein with increasing tumor grade, which is opposite to MDR1 expression. In addition, an opposite co-expression pattern of ERβ and individual MDR1 proteins was also observed. In conclusion, the results can be used to better understand the expression control of MDR1 and may allow for the establishment of new cancer chemistry strategies that will control P-gp expression in NSCLC.

Comparative study of estrogen receptor α, β mRNA expressions of endometriosis and normal endometrium in women and analysis of potential synthetic anti-estrogens in silico

Endometriosis is a multifactorial disease in which genetic and environmental factors interact causing its pathogenesis. The aim of this study was to investigate the expression pattern of estrogen receptor α (ERα) and β (ERβ) in endometriosis patients compared to normal endometrioum (n=18) as a control by using Quantitative Real Time PCR method. Moreover, we also measured serum estradiol levels of endometriosis patients in the proliferation phase of the menstrual cycle using the enzyme-linked immunosorbent assay method. The mRNA expression of ERβ was significantly higher in the endometriosis group compared to control, and the result of t-test showed that were significantly different (P&lt;0.05). Overexpression of ERβ in endometriosis was likely to have other significant important impacts in the pathology of endometriosis that allowed ERβ to stimulate prostaglandin production in endometriosis tissue and cells. Estradiol content did not correlate with the ERα expression, and it is weakly correlated with ERβ mRNA expression. Molecular docking analysis showed that ERα and ERβ have different binding interactions with synthetic antiestrogens, whereas the best inhibitor was Ral2 to ERα and Aco1 to ERβ. Thus, both inhibitors could be used as leads in further investigation of ERα, ERβ inhibitory activities in vitro and in vivo.

Loss of Angelman Syndrome Protein E6AP Disrupts a Novel Antagonistic Estrogen-Retinoic Acid Transcriptional Crosstalk in Neurons.

Angelman syndrome (AS) is a complex genetic disorder that affects the nervous system. AS affects an estimated 1 in 12,000 to 20,000 individuals. Characteristic features of AS includes developmental delay or intellectual disability, severe speech impairment, seizures, small head size (microcephaly), and problems with movement and balance (ataxia). AS individuals usually have microdeletion of the maternal copy of 15q11.2-15q13 region of chromosome 15. The E6-associated protein (E6AP, an E3 ubiquitin protein ligase enzyme) is encoded by the gene UBE3A, which is located in this region, and it has been shown that deregulation of E6AP gives rise to AS and neuropathology of autism spectrum disorders (ASDs) (e.g., autism and Rett syndromes). We have shown that E6AP also acts as a coactivator of the estrogen receptor (ER). ER is a ligand-induced transcription factor that exerts potent and wide-ranging effects on the developing brain. Furthermore, the expression pattern of ER in the brain overlaps with that of E6AP. Up till now, all the published studies have examined the role of the ubiquitin-protein ligase activity of E6AP in the development of AS, and it is not known what role the newly discovered coactivation functions of E6AP and ER plays in the pathology of AS. Here, we demonstrate that E6AP and ER co-immunoprecipitate and are in the same protein complex in neuronal cells (Neuro2a). In addition, both colocalize in nuclear and cytoplasmic compartments of the mouse hippocampal neurons and Neuro2a cells. Moreover, we identified a novel E6AP and ER direct transcriptional regulation of a gene Cyp26b1 known to be involved in learning and memory processes. This transcriptional regulation involves recruitment of E6AP and ER to a newly discovered functional estrogen response element (ERE) located at the Cyp26b1 gene promoter and is associated with transcription permissive epigenetic events leading to increase of active transcription of the gene in neurons upon estrogen treatment. This novel transcriptional regulation was also validated in the AS mouse model where E6AP expression is abrogated in the mouse brain. In fact, Cyp26b1 expression is decreased by 31% in AS mice versus age-matched control (Ctrl) mice hippocampi. Also, retinoic acid transcriptional signaling was shown to be amplified as evidenced by specific increased Rarβ and decreased Erbb4 mRNA expression in AS mice versus Ctrl mice hippocampi. These transcript level changes were also supported by the same trend of changes at the protein level. Collectively, our data present a proof of principle that the transcriptional coactivation function of E6AP may have a crucial role in the pathobiology of AS. This function, yet to be thoroughly investigated, reveals the possibility of harnessing the antagonistic estrogen-retinoic acid transcriptional signaling crosstalk and potentially other unknown effectors for the investigation of important possible targets as putative novel treatment modalities and venues for reversing neurological manifestations in AS and related syndromes like ASDs.

Sex hormone levels and expression of their receptors in lactating and lactating pregnant rats

Parturient rats show a postpartum estrus, a period of sexual receptivity that occurs from 6 to 15 h after the birth of a litter, which allows the mother to gestate a second litter while simultaneously nursing the first one (lactating and pregnant). The present study investigated hormone levels and the expression pattern of estrogen receptor α, and β, progesterone receptor isoforms and SRC1 in the hypothalamus and the preoptic area of lactating as well as in lactating-pregnant rats. In the latter, estradiol levels were 3-fold higher than those observed in lactating rats on day 14, meanwhile progesterone levels did not change in any condition. There were higher levels of prolactin in both lactating and lactating-pregnant rats on day 7 and decreased on the following days. In the hypothalamus of the lactating rat, the content of ERα increased during lactation meanwhile that of ERβ decreased 50% on day 10. The content of both estrogen receptor subtypes in the hypothalamus increased 3-fold on day 21 in lactating-pregnant rats. In the preoptic area, the content of ERα was higher in lactating-pregnant rats on days 14 and 21 while the content of progesterone receptor isoforms was lower as compared with those found in lactating animals on days 7 and 10. The content of SRC1 increased 2-fold in the preoptic area only in lactating rats at day 14 and 21. These findings suggest that lactating- pregnant animals should exhibit differential neuroendocrine and molecular characteristics as compared to lactating animals.

Temporal expression and gonadotropic regulation of aromatase and estrogen receptors in the ovary of wall lizard, Hemidactylus flaviviridis: Correlation with plasma estradiol and ovarian follicular development

The current study in Indian wall lizard Hemidactylus flaviviridis for the first time demonstrates the reproductive phase-dependent expression pattern of aromatase (cyp19) and estrogen receptor subtypes (er-α and er-β) as well as their gonadotropic regulation in the ovary of a squamate. The expression of cyp19 remained low during regressed phase, increased markedly in recrudescent and declined sharply in breeding phase. Further, temporal profile of plasma estradiol 17-β (E2) was found to be relatively parallel to the expression pattern of ovarian cyp19. The expression pattern of estrogen receptors in the ovary showed subtype-specific variation along the reproductive cycle. Expression of ovarian er-α remained high from regressed to late recrudescence, while er-β expression that was low during regression dramatically increased with the initiation of follicular growth in early recrudescence and remained high until late recrudescence. Nonetheless, expression of both the receptors declined during breeding phase when ovary contained vitellogenic follicle. Regarding gonadotropic regulation, short-term treatment with Follicle stimulating hormone (3 injections of FSH) increased the ovarian expression of cyp19, er-α and er-β while prolongation of treatment (7 or 11 injections) resulted in a marked decrease in expression of these genes concomitant to formation of vitellogenic follicle. However, a marked increase in plasma E2 was recorded after 7 injections of FSH. The direct role of gonadotropin in regulation of cyp19 and estrogen receptors was established by an in vitro study where FSH upregulated the expression of these genes in all stages of ovarian follicles (early growing, previtellogenic and early vitellogenic) of wall lizards.

The Effect of Histone Hyperacetylation on Viability of Basal-Like Breast Cancer Cells MDA-MB-231

Abstract Background: The Basal-Like breast cancer, is always known for lack of expression of estrogen receptor (ER), progesterone receptor (PR) and as well, absence of epidermal growth factor receptor 2 (HER2) gene amplification. Improper expression pattern of ER, PR, and Her2, makes Basal-Like breast tumors resistant to the current hormonal and anti HER2 treatments. In recent decades, several studies have been conducted to investigate the regulatory role of chemical modifications of core histones in gene expression. Their results have shown that histone acetylation is involved in regulation of cell survival. Acetylation of core histones is regulated by the epigenetic-modifying enzymes named Histone Deacetylases (HDACs). As a new approach to control the viability of breast tumor cells resistant to the hormonal and anti-HER2 treatments, we have targeted the HDACs. Using Trichostatin A (TSA) as a known HDACs inhibitor, we have tried to hyperacetylate the core histones of MDA-MB-231 cells as an in vitro model of Basal-Like breast tumors. Then we have investigated the effect of histone hyperacetylation on viability of MDA-MB-231 cells. Methods: MDA-MB-231 cells were cultured in RPMI 1640 medium containing 10% fetal bovine serum (FBS) and were incubated at 37°C, in a humidified incubator with 5% CO2 atmosphere. Then cells were treated with different concentrations of TSA including: 50, 100, 200, 400, 800 and 1000 nM or control (1% DMSO). After 24 and 48 hours, viability of cells was evaluated by MTT assay. Results: After 24 and 48h exposure to different concentrations of TSA, MDA-MB-231 cells showed a maximum tolerable dose. At higher concentrations, TSA decreased the percentage of cell viability through a time-dose dependent manner. IC50 value for 48h treatment was 600 nM. Conclusions: Our results indicate that HDACs inhibition and subsequently hyperacetylation of histones, leads to cytotoxic effects on breast tumor cells resistant to the current treatments. Following this pilot research we are trying to suggest molecular mechanisms underlying the anti-proliferative effects triggered by HDACs inhibition.

Study on Estrogen Receptor, Progesterone Receptor and HER-2/ neu Expression Pattern by Immunohistochemistry in 87 cases of Invasive Breast Cancer

Background: Now a day's determination of estrogen receptor (ER), progesterone receptor (PR) and HER-2/neu expression pattern by immunohistochemistry in invasive breast cancer have become the standard procedure for breast cancer management.Objective: To see the expression pattern of estrogen receptor, progesterone receptor and HER-2/neu in Bangladeshi women with invasive breast carcinoma.Method: This cross sectional study was performed in 87 cases of invasive breast cancer. Estrogen receptor (ER), Progesterone receptor (PR) and HER-2/neu expression pattern were assessed by immunohistochemistry using monoclonal antibodies for detecting estrogen and progesterone receptors, and polyclonal antibody for detecting HER-2/neu.Results: All the cases were graded according to Bloom-Richardson grading system. Of those, Grade I tumour was 18 (20.69%), Grade II tumour was 58 (66.67%) and Grade III tumour was 11(12.64%). Both ER and PR positive reactivity were same and it was found 65 (74.71%) and HER-2/neu reactivity pattern were found negative in 59 (67.82%) cases and positive in 28 (32.18%) cases. A statistically significant correlation was found between the expression of ER and low grade tumour (p=0.011) and combined estrogen and progesterone receptor positive reactivity with low grade tumour (p=0.002).Conclusion: ER, PR and HER-2/neu expression do not correlated with each other, so it is recommended that each test should be independently determined by immunohistochemistry in all cases of invasive breast cancer. All equivocal cases of HER-2/neu (score 2+) should be analyzed by FISH technique to find out the percentage of real score.KYAMC Journal Vol. 5, No.-1, Jul 2014, Page 436-443

Molecular expression of caprine estrogen receptor gene 1 in reproductive and non-reproductive tissues.

During the last decades, physiological effects of oestrogens have been increasingly explored by scientists and biotechnologists. Estrogens exert a wide range of effects on a large variety of cell types. Oestrogen and its receptors are essential for sexual development and reproduction. Estrogen receptor alpha is a nuclear receptor activated by the hormone oestrogen. In male, ERα is encoded by the gene estrogen receptor gene 1 (ESR1), responsible for better fertility. The ESR1 is involved in the reabsorption of luminal fluid during the transit of spermatozoa from the testis to the head of the epididymis which is important for their survival and maturation during epididymal storage. The absence of ESR1 leads to reduced epididymal sperm content, reduced sperm motility and fertilizing ability. Therefore, this is a good startby to study the expression pattern of estrogen receptor 1 gene in high-fertile (G1) and low-fertile (G2) bucks of Jamunapari and Barbari breeds identified on the basis of seminal quality traits and fertility trials. RNA was extracted from the tissues by TRIzol method. The identification and expression pattern of caprine ESR1 gene was analysed by real-time PCR (Roche LC-480). Our work shows that the relative quantification by RT-PCR indicates more fold in head of epididymis as compared to spleen of caprine ESR1 gene. Furthermore, the RT-PCR indicated that fertile bucks of Jamunapari breed have more fold value as compared to Barbari breed in respect of reproductive organ.

Estrogen receptor gene 1 expression in caprine and its effect on fertility.

The present study was undertaken to analyze the expression pattern of estrogen receptor 1 gene (ESR1) in Barbari bucks (fertile and non-fertile) identified on the basis of seminal quality traits and fertility trials. RNA was extracted from the spleen by Trizol method. The expression pattern of ESR1 gene was analyzed using real time polymerase chain reaction (RT-PCR). The expression pattern of ESR1 gene was analyzed by RT-PCR (Roche LC-480). Relative quantification by RT-PCR indicated that the ESR1 gene expression showed more fold in fertile bucks as compared to non-fertile.

Hormonal Receptor, Human Epidermal Growth Factor and Its Association with Breast Cancer Tumor Characteristics in Albania.

This retrospective study was designed to analyze expression patterns of estrogen receptor (ER), progesterone receptor (PR) and HER2/neu in Albanian patients with breast carcinoma to identify their relationships with tumor size, histological grade (HG), lymph node metastasis and relapse. Patients with either biopsy or metastatic relapse were identified. Demographics, tumor characteristics, ER, PR, and HER2/neu status were retrospectively obtained from the medical records of patients treated with breast cancer during 2006-2011. Hormonal receptors and HER2/neu were assessed by immunohistochemistry. Association of ER, PR and HER2/neu with clinicopathological and molecular characteristics were studied using Fisher's test. P value ≤0.05 was considered significant. There were 110 patients included in the study. Mean patient age was 51.08±10.75 years. The overall immunoexpression of ER, PR and HER2/neu were found positive in 76 (69%), 73 (67%), and 16 (41%) patients, respectively. ER- was associated with higher histological grade (24% vs. 9.2%) and PR+ with tumor size (T2, 78.3 vs. 64.3) (p=0.02 and 0.05, respectively). ER and PR expression were significantly decreased in HER2/neu positive cases while HER2/neu levels correlated with tumor size (p=0.03) and nodal metastasis (p=0.03). No association was detected between ER, PR, HER2/neu and relapse. A combination of ER, PR and HER2/neu and prognostic factors could be of clinical value by defining subgroups in Albanian breast cancer patients that might benefit from more aggressive treatment.