High ERβ Expression Research Articles

Cisplatin-activated ERβ/DCAF8 positive feedback loop induces chemoresistance in non-small cell lung cancer via PTEN/Akt axis

High levels of the estrogen receptor β (ERβ) predict poor prognosis following platinum-containing adjuvant chemotherapies in patients with non-small cell lung cancer (NSCLC). However, the precise role of ERβ remains elusive. In this study, we demonstrated that targeting ERβ could significantly increase the cytotoxicity of cisplatin both in vitro and in vivo. Mechanically, cisplatin directly binds to ERβ, which facilitates its homodimerization and nuclear translocation. ERβ activation transcriptionally represses the expression of DCAF8, an adaptor of CRL4 E3 ubiquitin ligase, which in turn attenuates the proteasomal degradation of ERβ, leading to ERβ accumulation; this positive feedback loop results in Akt activation and eventually cisplatin resistance in NSCLC through PTEN inhibition. Moreover, low expression of DCAF8 and high expression of ERβ are associated with treatment resistance in patients receiving cisplatin-containing adjuvant chemotherapy. The present results provide insights into the underlying mechanism of ERβ-induced cisplatin resistance and offer an alternative therapeutic strategy to improve the efficacy of platinum-based chemotherapy in patients with NSCLC.

Association of the Estrogen Receptors Beta Expression with the Ki-67 Proliferative Index in Breast Cancer

Background. Estrogen receptors beta (ERβ) are an important biological regulator and target of antiestrogens, however, unlike estrogen receptors alpha (ERɑ), their significance in the prognosis and treatment of breast cancer remains unclear. Purpose. Evaluation of the ERβ prognostic value in the comparative assessment of frequency and level of the marker expression in groups with good and poor prognosis by Ki-67 proliferative index score in breast cancer. Methods. ERβ expression level (% of cells expressing the marker) in 68 breast tissue samples was quantified by immunofluorescence and flow cytometry. Primary antibodies to ERβ (clone 14C8, ab288) and secondary antibodies conjugated with DyLight650 (ab98729) were used. In the same samples, the Ki-67 expression level was assessed by the immunohistochemical method. Results. The ERβ and Ki-67 were detected in 100% breast tissue samples with high heterogeneity of the markers’ expression in different patients. Statistical analysis of good and poor prognosis in accordance with the Ki-67 proliferative index score (Ki-67≤20% and Ki-67>20%) showed the prognostic value of the ERβ expression level of 50%. There was no association between the Ki-67 and ERβ expression levels in the same tumor sample (Spearman's rank correlation coefficient R=–0,16; P>0,05). At the same time, high expression of ERβ≥50% was 2,3 times more frequently detected in the good vs poor prognostic group by Ki-67 — 41% vs 18%, P=0,02. Conclusion. The ERβ expression level ≥50% in the tumor can be considered as a factor of good prognosis of breast cancer.

Estrogen Receptor-β Gene Cytosine-Adenine (ESR2-CA) Repeat Polymorphism in Postmenopausal Colon Cancer

The pathobiological role of estrogen is controversial in colorectal cancer. Cytosine-adenine (CA) repeat in the estrogen receptor (ER)-β gene (ESR2-CA) is a microsatellite, as well as representative of ESR2 polymorphism. Though its function is unknown, we previously showed that a shorter allele (germline) increased the risk of colon cancer in older women, whereas it decreased it in younger postmenopausal women. ESR2-CA and ER-β expressions were examined in cancerous (Ca) and non-cancerous (NonCa) tissue pairs from 114 postmenopausal women, and comparisons were made considering tissue types, age/locus, and the mismatch repair protein (MMR) status. ESR2-CA repeats <22/≥22 were designated as 'S'/'L', respectively, resulting in genotypes SS/nSS (=SL&LL). In NonCa, the rate of the SS genotype and ER-β expression level were significantly higher in right-sided cases of women ≥70 (≥70Rt) than in those in the others. A decreased ER-β expression in Ca compared with NonCa was observed in proficient-MMR, but not in deficient-MMR. In NonCa, but not in Ca, ER-β expression was significantly higher in SS than in nSS. ≥70Rt cases were characterized by NonCa with a high rate of SS genotype or high ER-β expression. The germline ESR2-CA genotype and resulting ER-β expression were considered to affect the clinical characteristics (age/locus/MMR status) of colon cancer, supporting our previous findings.

Targeting circDGKD Intercepts TKI's Effects on Up-Regulation of Estrogen Receptor β and Vasculogenic Mimicry in Renal Cell Carcinoma.

Simple SummaryOur aim was to elucidate the molecular mechanisms of how tyrosine kinase inhibitors, including sunitinib, contribute to vasculogenic mimicry (VM) formation and progression in renal cell carcinoma. We demonstrated that sunitinib and axitinib could induce ERβ expression in RCC cell lines and ERβ transcriptionally up-regulated the circular RNA of DGKD (circDGKD, hsa_circ_0058763) expression. The circDGKD could sponge tumor suppressor miR-125-5p family members and consequently led to increased VE-cadherin, the key adhesion molecule in the VM formation process, which is targeted by miR-125-5p at the 3′ UTR, providing novel targets for combination therapy in clinical metastatic RCC patients.Vasculogenic mimicry (VM) has been reported as an alternative channel to increase tumor nutrient supplies and accelerate tumor progression, and is associated with poor survival prognosis in multiple cancers, including renal cell carcinoma (RCC). The currently used anti-angiogenic treatment for metastatic RCC, sunitinib, a tyrosine kinase inhibitor (TKI), has been reported to induce VM formation. Previously we identified that the estrogen receptor β (ERβ) functions as an oncogenic factor to promote RCC progression, supported by the analytic results from The Cancer Genome Atlas (TCGA) database. We have also found evidence that sunitinib induces RCC VM formation by up-regulating ERβ expression. In this study, we further demonstrated that treatment with sunitinib, as well as axitinib, another TKI, could induce ERβ expression in RCC cell lines. Clinical clear cell RCC (ccRCC) patients with higher ERβ expression are more likely to be found VE-cadherin positive and VM positive. Mechanism dissection showed that TKI- induced ERβ transcriptionally up-regulates the circular RNA of DGKD (circDGKD, hsa_circ_0058763), which enhances VE-cadherin expression by sponging the microRNA miR-125-5p family. Targeting circDGKD intercepts sunitinib-pretreatment-induced RCC VM formation, reduces metastases and improves survival in an experimental orthotopic animal model. Targeting ERβ/circDGKD signals may improve the TKI efficacy and provide novel combination therapies for metastatic RCC.

Upregulation of estrogen receptor beta protein but not mRNA predicts poor prognosis and may be associated with enhanced translation in non-small cell lung cancer: a systematic review and meta-analysis.

BackgroundAn increasing number of original studies suggest that estrogen receptor beta (ERβ) expression may be related to non-small cell lung cancer (NSCLC) prognosis; however, the evidence remains inconclusive and conflicting. We aimed to systematically evaluate the expression and prognostic value of ERβ in NSCLC, and to explain the inconsistency between ERβ protein and mRNA level.MethodsPubMed, Embase, and Web of Science databases were searched for studies (published before October 6, 2020) reporting the prognostic value of ERβ protein expression in NSCLC. The pooled hazard ratios (HRs) with 95% confidence intervals (CIs) for overall survival (OS) were calculated. Transcriptome and survival data of lung adenocarcinoma patients were obtained from public databases for differential expression and survival analyses. Immunohistochemistry (IHC) was performed to examine the ERβ protein expression in 39 NSCLC patients. Western blotting and RT-qPCR were performed to analyze ERβ expression in two paired NSCLC and normal adjacent tissue samples. The effect of methyltransferase-like 13 (METTL3) on ERβ expression was investigated in a lung cancer cell line.ResultsMeta-analysis of 23 studies with a total of 3744 patients demonstrated that high protein expression of overall ERβ and cytoplasmic ERβ indicated poor OS (HR: 1.05, 95% CI: 1.00 to 1.10; HR: 1.48, 95% CI: 1.13 to 1.95) in NSCLC. For lung adenocarcinoma especially, high protein expression of both overall/cytoplasmic ERβ and nuclear ERβ suggested poor OS (HR: 1.54, 95% CI: 1.05 to 2.25; HR: 1.36, 95% CI: 1.03 to 1.80). Bioinformatics analysis indicated the expression of ERβ mRNA was not associated with the prognosis of lung adenocarcinoma. Analysis of public databases showed that ERβ mRNA is not highly expressed in tumor tissues, however, IHC results revealed that ERβ protein is highly expressed in NSCLC tissues. We validated this inconsistency in ERβ expression in paired tumors and normal adjacent tissues from patients. Moreover, METTL3 knockdown in the A549 cell line downregulated ERβ protein expression but not ERβ mRNA expression.ConclusionsOur study elucidated the inconsistency between ERβ protein and mRNA expression levels and their prognostic values. The results indicated that METTL3-driven enhanced translation in NSCLC may cause this inconsistency.

Antitumor effects of aconitine in A2780cells via estrogen receptor β‑mediated apoptosis, DNA damage and migration.

Ovarian cancer (OVCA) is the deadliest type of malignant gynecological disease, and previous studies have demonstrated that estrogen receptor β (ERβ) serves important roles in this disease. Aconitine, a toxin produced by the Aconitum plant, displays potent effects against cancers. The aim of the study was to investigate the pharmacological activities and mechanisms of aconitum on OVCA. In the present study, the activity of aconitine in the human OVCA A2780 cell line was investigated. The results revealed that aconitine suppressed cell viability, colony formation and motility. Terminal deoxynucleotidyl-transferase-mediated dUTP nick end labeling, mitochondria membrane potential and comet assays showed that aconitine induced mitochondria apoptosis and DNA damage in A2780 cells. Investigation of the mechanism revealed that a high expression of ERβ and prolyl hydroxylase 2 was detected after aconitine treatment, and aconitine significantly suppressed the expression of vascular endothelial growth factor and hypoxia-inducible factor 1α to activate ERβ signaling. Moreover, the expression levels of p53, Bax, apoptotic peptidase activating factor 1, cytochrome C, cleaved caspase-3/9 and cleaved poly (ADP-ribose) polymerase were upregulated, and the expression levels of Bcl-2, Bcl-xl and phosphorylated ATM serine/threonine kinase were downregulated by aconitine. Interestingly, aconitine also markedly downregulated the expression of matrix metalloproteinase 2 (MMP2) and MMP9, which are associated with tumor invasion. In addition, a molecular docking assay revealed that aconitine exerted strong affinity towards ERβ mainly through hydrogen bonding and hydrophobic effects. Collectively, these results suggested that aconitine suppressed OVCA cell growth by adjusting ERβ-mediated apoptosis, DNA damage and migration, which should be considered a potential option for the future treatment of OVCA.

Tumour-suppressive effect of oestrogen receptor β in colorectal cancer patients, colon cancer cells, and a zebrafish model.

Oestrogen receptor β (ERβ) has been suggested to have anti-proliferative and anti-tumour effects in breast and prostate cancer cells, but other studies have indicated its tumour-promoting effects. Understanding the complex effects of this receptor in different contexts requires further study. We reported that high ERβ expression is independently associated with improved prognosis in female colorectal cancer (CRC) patients. Herein, we investigated the possible anti-tumour effect of ERβ and its selective agonist. CRC patients with high ERβ expression had significantly higher levels of membrane-associated β-catenin, cysteinyl leukotriene receptor 2 (CysLT2 R), and 15-hydroxyprostaglandin dehydrogenase (15-PGDH), which have anti-tumour effects, but lower levels of nuclear β-catenin, cysteinyl leukotriene receptor 1 (CysLT1 R), and cyclooxygenase-2 (COX-2), which have tumour-promoting effects. These interesting findings were further supported by two different publicly available CRC mRNA datasets that showed a significant positive correlation between ERβ expression and 15-PGDH and CysLT2 R expression and a negative correlation between ERβ expression and β-catenin, CysLT1 R, and COX-2 expression. We next evaluated ERβ expression in three different colon cancer mouse models; ERβ expression was negatively correlated with tumourigenesis. Furthermore, treatment with the ERβ-agonist ERB-041 reduced CysLT1 R, active β-catenin, and COX-2 levels but increased phospho-β-catenin, CysLT2 R, and 15-PGDH levels in HCT-116, Caco-2, and SW-480 colon cancer cells compared to vehicle-treated cells. Interestingly, ERB-041-treated cells showed significantly decreased migration, survival, and colonosphere formation and increased apoptotic activity, as indicated by increased CASPASE-3 and apoptotic blebs. Finally, patients with low ERβ expression had significantly more distant metastasis at the time of diagnosis than patients with high ERβ expression. Therefore, we studied the effects of ERB-041-treated colon cancer cells in a zebrafish xenograft model. We found significantly less distant metastasis of ERB-041-treated cells compared to vehicle-treated cells. These results further support ERβ's anti-tumour role in CRC and the possible use of its agonist in CRC patients. © 2020 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland.

Abstract A96: Correlation of estrogen receptor beta with immune cells in colon cancer patients

Abstract Background: The estrogen receptor β (ERβ) is the predominant ER in the colon mucosa. We have previously reported that high expression of ERβ is independently associated with a better prognosis in patients with colorectal cancer (CRC), and the induction of this receptor in colon cancer cell lines resulted in upregulation of antitumorigenic proteins and downregulation of protumorigenic proteins. In addition, we have also shown that high mast cell density (MCD) was associated with longer survival of CRC patients. Aim: To investigate the correlations between ERβ, mast cells (MCs), and prostaglandin D2 (PGD2) synthase. Materials and Methods: A tissue microarray (TMA) of primary CRCs from 314 patients was stained for the expression of ERβ, MCs, and PGD2 synthase. Immunohistochemistry was used to evaluate the staining intensity by using the immunoreactive score (IRS). Results: Patients with high ERβ expression had higher number of MCs, and there was a significant positive correlation between ERβ and MCs (r = 0.12, P = 0.03). In addition, patients with high ERβ expression had significantly higher activity of PGD synthase, which is produced mainly from MCs and is known to have antitumor effects. Furthermore, a strong, positive, and significant correlation was observed between ERβ expression and PGD synthase (r = 0.42, P &amp;lt; 0.0001). Future Plans: We aim to correlate the expression of ERβ with other immune cells and factors important for the tumor microenvironment, such are CD3+, CD8+, T-bet, and PDL-1. Conclusion: High ERβ expression correlates with an antitumor immune prolife in CRC patients. This strengthens our hypothesis that the induction of ERβ expression in CRC patients will potentially improve the disease outcome. Citation Format: Geriolda Topi, Marie Louise Lydrup, Anita Sjölander. Correlation of estrogen receptor beta with immune cells in colon cancer patients [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2019 Nov 17-20; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2020;8(3 Suppl):Abstract nr A96.

The prognostic values of estrogen receptor alpha and beta in patients with gastroesophageal cancer: A meta-analysis.

Published studies have investigated the prognostic roles of estrogen receptor alpha (ERα) and estrogen receptor beta (ERβ) in gastroesophageal cancer patients with the controversial results. The aim of the study was to systematically evaluate the impacts of ERα and ERβ on the overall survival (OS) in patients. Relevant eligible studies were extracted from PubMed, Embase, Web of Science, CNKI and Wanfang databases (from the start date to November 2018) following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. HR (hazard ratio) with 95% confidence intervals (CIs) were used to assess the prognostic values of ERα and ERβ for OS in patients. High ERα expression was associated with poor OS (HR = 1.58, 95% CI = 1.29-1.94, P < .001) and ERβ with better OS (HR = 0.56, 95% CI = 0.37-0.83, P = .004) in gastroesophageal cancer. Furthermore, unfavorable OS was found in Chinese gastroesophageal patients with higher ERα expression (HR = 1.57, 95% CI = 1.25-1.96, P < .001) and better OS with higher ERβ expression (HR = 0.51, 95% CI = 0.31-0.83, P < .01) in our subgroup analysis. Meanwhile, worse OS was found in esophageal squamous cell carcinoma (ESCC) patients with high ERα expression (HR = 1.74, 95% CI = 1.33-2.26, P < .001), and favorable OS in ESCC with ERβ overexpression (HR = 0.40, 95% CI = 0.31-0.52, P < .001). Besides, high ERα expression was associated with lower tumor differentiation in ESCC (OR = 1.64; 95% CI = 1.02-2.64, P = .04) and ERβ was linked with better tumor differentiation in gastric adenocarcinoma (GCA) (OR = 0.49; 95% CI = 0.26-0.94, P = .03). ERα and ERβ might serve as potential prognostic biomarkers for gastroesophageal cancer patients. ERα overexpression predicted poor OS and lower tumor differentiation, and ERβ suggested favorable OS and better tumor differentiation. Further related studies should be performed to test these results.

Overexpression Of ERβ Participates In The Progression Of Liver Cancer Via Inhibiting The Notch Signaling Pathway.

PurposeThis study aimed to explore the role of Estrogen Receptor-β (ERβ)-mediated Notch signaling pathway in the regulation of proliferation and apoptosis in liver cancer cells.MethodsHepG2 cells (Pbi-EGFP-ER) were transfected with ERβ that mediated by liposome, and normal HepG2 cells (Blank) and empty plasmid-transfected HepG2 cells (Pbi-EGFP-C) were used as controls. Then, Huh7 cells were transfected with shERβ lentivirus to knock down ERβ expression. The Huh7 cells were divided into three groups including Blank, experimental group (shERβ) and negative group (shLuc). Then, qRT-PCR, Western blot, CCK-8 assay, cell scratch assay, Transwell assay, Annexin V-FITC and PI double staining were performed based on these groups. Finally, a mouse xenograft model was constructed to verify the regulation of ERβ on Notch signaling pathway in liver cancer.ResultsIn HepG2 cells, the ERβ expression in Pbi-EGFP-E group was higher than that in Blank and Bi-EGFP-C group. Overexpression of ERβ inhibited HepG2 cell proliferation, migration, invasion and Ki67 protein expression, as well as promoted apoptosis, Bcl-2 and Bax expression. Overexpression of ERβ decreased Notch1, Notch2 and Hes1 expression. In Huh7 cells, the effect of low ERβ expression was contrary to that of high ERβ expression. The shERβ + DAPT group reversed the effect of shERβ on the volume and weight of transplanted tumors.ConclusionERβ may inhibit the development of liver cancer and promote apoptosis via inhibiting the Notch pathway.

Abstract 5032: High ERbeta expression correlates with the tumor suppressor 15-PGDH and PGD2 synthase in a female cohort of colorectal cancer patients

Abstract Background: The estrogen receptor β (ERβ) is the predominant ER in the colon mucosa. We have previously reported that high expression of ERβ is independently associated with a better prognosis in female patient with colorectal cancer (CRC). β-catenin and cyclooxygenase-2 (COX-2) are well known to be involved in CRC tumorigenesis. We have shown that a high level of CysLT2R, a low level of CysLT1R and high mast cell density are correlated with good prognosis in CRC patients. β-catenin and some inflammatory mediators, such are cyclooxygenase-2 (COX-2), cysteinyl leukotriene receptor 1 and 2 (CysLT1R, CysLT2R), prostaglandin D2 (PGD2), 15-PGDH activity and mast cells have been associated with CRC. Aim: To investigate the association between ERβ and other tumor modulators in CRC patients, as well as the underlying mechanism in colon cancer cell lines. Material and methods: A tissue microarray of primary CRCs from 320 female patients was stained for the expression of ERβ, COX-2, 15-PGDH, CysLT1R, CysLT2R, ß-catenin, PGD synthase and mast cells. The Immunohistochemistry technique was used to evaluate the staining intensity, which was assigned an Immuno-Reactive Score (IRS). The zebrafish xenograft model was used to study colon cancer cells metastasis. ERβ -041 (an ERβ agonist) was injected into the zebrafish perivitelline space to allow for xamination of its effect on colon cancer cells. After 48 h, the tail, considered the distant metastatic site, was investigated for the presence of the metastasized cells. The effect of ERβ on colon cancer cells (SW-480 and HCT-116) after treatment with ERβ-041 was studied by wound healing and trans-well migration assay, as well as by survival assay. Results: Patients with high ERβ expression had significantly higher levels of membrane ß-catenin, CysLT2R, 15-PGDH and PGD synthase, which all are known to have an anti-tumor effect. These patients had significantly lower levels of CysLT1R, COX-2 and nuclear ß-catenin expression, which are known to have pro-tumorigenic effects. In addition, patients with high ERβ expression had higher numbers of mast cells, which are a major source of PGD2 synthase and PGD2. In the zebrafish xenograft model, the ERB-041 treated cells showed no distant metastasis, unlike the untreated cells. ERβ stimulation of colon cancer cells produced a significant decrease in migration and survival compared to that of the untreated control cells after 48 h post-stimulation. Conclusion: High ERβ expression was significantly correlated with anti-tumorigenic inflammatory mediators and lower nuclear ß-catenin in patients with CRC. These results strengthen our hypothesis regarding the beneficial role of ERβ in CRC patients. Our results raise the possibility of cross-talk between ERβ, CysLT1R and CysLT2R signalling. Citation Format: Geriolda Topi, Pujarini Dash, Shakti R. Satapathy, Syrina Mehrabi, Roy Ehrnström, Marie-Louise Lydrup, Anita Sjölander. High ERbeta expression correlates with the tumor suppressor 15-PGDH and PGD2 synthase in a female cohort of colorectal cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 5032.

Estrogen receptor β upregulates CCL2 via NF-κB signaling in endometriotic stromal cells and recruits macrophages to promote the pathogenesis of endometriosis.

How is the activation of estrogen receptor β (ERβ) in endometriotic stromal cells (ESCs) involved in macrophage recruitment to promote the pathogenesis of endometriosis? ERβ modulates the production of C-C motif chemokine ligand 2 (CCL2) via nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling in ESCs and thus recruits macrophages to ectopic lesions to promote pathogenesis. Macrophages are mainly recruited to the peritoneal cavity to promote the pathogenesis of endometriosis. Recent studies have demonstrated that ERβ plays an important role in the progression of endometriosis through modulating apoptosis and inflammation. An observational study consisting of 22 cases (women with endometriosis, diagnosed by laparoscopy and histological analysis) and 14 controls (without endometriosis) was carried out. Tissues and stromal cells that were isolated from 22 patients with ovarian endometrioma and deeply infiltrating endometriosis were compared with tissues and stromal cells from 14 patients with normal cycling endometrium using immunohistochemistry, quantitative PCR, Western blot, cell migration assay and cloning formation assay. P values < 0.05 were considered significant, and experiments were repeated in at least three different cell preparations. We observed that accumulated macrophages were recruited to the ectopic milieu and mainly adopted an alternatively activated macrophage (M2) phenotype. To reveal the underlying mechanism for this, we conducted a series of experiments and found that high expression of ERβ led to the production of CCL2 via NF-κB signaling and macrophages were recruited to the ectopic milieu. An in vitro co-culture assay also suggested that the recruited macrophages in turn could promote the proliferation and clonogenic ability of ESCs. Overall, the activation of ERβ in ESCs is involved in macrophage recruitment via NF-κB/CCL2 signaling and subsequently appears to promote the pathogenesis of endometriosis. N/A. Due to the limitations of obtaining surgical specimens, endometrioma tissues were collected mainly from women diagnosed with middle to late stage endometriosis. We identified the predominant presence of M2 macrophages in the samples used in our study, but the underlying mechanism of how recruited macrophages acquire the M2 phenotype is undefined. This work provides novel insight into the mechanism by which ERβ may modulate macrophage infiltration and promote pathogenesis, which may provide a new therapeutic target for endometriosis. This study was supported by the National Natural Science Foundation of China (81671430). The authors have no conflicts of interest.

G-Protein-Coupled Estrogen Receptor Antagonist G15 Decreases Estrogen-Induced Development of Non-Small Cell Lung Cancer.

G-protein-coupled estrogen receptor (GPER) was found to promote non-small cell lung cancer (NSCLC) by estrogen, indicating the potential necessity of inhibiting GPER by a selective antagonist. This study was performed to elucidate the function of GPER-selective inhibitor G15 in NSCLC development. Cytoplasmic GPER (cGPER) and nuclear GPER (nGPER) were detected by immunohistochemical analysis in NSCLC samples. The relation of GPER and estrogen receptor β (ERβ) expression and correlation between GPER, ERβ, and clinical factors were analyzed. The effects of activating GPER and function of G15 were analyzed in the proliferation of A549 and H1793 cell lines and development of urethane-induced adenocarcinoma. Overexpression of cGPER and nGPER was detected in 80.49% (120/150) and 52.00% (78/150) of the NSCLC samples. High expression of GPER was related with higher stages, poorer differentiation, and high expression of ERβ. The protein level of GPER in the A549 and H1793 cell lines was increased by treatment with E2, G1 (GPER agonist), or fulvestrant (Ful; ERβ antagonist) and decreased by G15. Administration with G15 reversed the E2- or G1-induced cell growth by inhibiting GPER. In urethane-induced adenocarcinoma mice, the number of tumor nodules and tumor index increased in the E2 or G1 group and decreased by treatment with G15. These findings demonstrate that using G15 to block GPER signaling may be considered as a new therapeutic target in NSCLC.

Estrogen receptor beta expression correlates with proliferation in desmoid tumors.

Estrogen receptor signaling and cyclin D1 have a major role in tumor cell proliferation in breast cancer. Desmoid tumors are rare neoplasms that may respond to endocrine treatment. The present study aimed to investigate the expression levels and the clinical relevance of estrogen receptor beta (ERβ) and cyclin D1 in desmoid tumors. This study consists of 83 patients with a surgically treated desmoid tumor. ERβ and cyclin D1 expression was examined by immunohistochemistry in tissue microarrays. Cyclin A and Ki67 were studied in our previous work. Median ERβ expression was 10.8%. ERβ expression correlated with expression of the proliferation antigens Ki67 (rp = 0.35, P = 0.003), cyclin D1 (rp = 0.34, P = 0.004), and cyclin A (rp = 0.34, P = 0.004). ERβ immunoexpression showed a trend towards predictive impact for recurrence as a continuous variable. Further explorative analysis indicated that very high ERβ expression was related to high risk of relapse (hazard ratio [HR] 2.6; P = 0.02). Median cyclin D1 expression was 15.6%. High cyclin D1 expression was associated with high Ki67 and cyclin A expression. Cyclin D1 was not associated with time to recurrence. ERβ and cyclin D1 immunopositivity correlated with high proliferation in desmoid tumors. High ERβ expression might be predictive for postoperative recurrence.

Level of estrogen receptor α and β expression and coexpression in non-small cell lung cancer tissue

Objectives. Quantitative assessment of the expression and co-expression levels of estrogen receptors of different types (ERα and ERβ) in malignant tissues of large cohort of patients with non-small cell lung cancer (NSCLC). Materials and methods . Measurement of ERα and ERβ expression levels was carried out on 167 malignant tissue samples by immunofluorescence flow cytometric method. Level of the ERα and ERβ expression was calculated as a ratio of specifically fluorescent cells (%) compared to the control (incubated with the secondary antibody only). Results. The expression of ERα and ERβ was detected in all the NSCLC samples investigated. High variability in the level of the marker expression was revealed and it was shown 1,5–2,0-fold higher ERβ expression in the most tumors than that for ERα. The Spearman rank correlation coefficient of 0,364 (p <0,001) and the Pearson correlation coefficient of 0,401 (p <0,001) indicate statistically significant weak positive correlation between the levels of expression of ERβ and ERα. Conclusion . The level of expression of ERβ as a main subtype of ER in NSCLC tissue could prove useful for selecting patients for antiestrogen therapy

Cyclooxygenase-2 and estrogen receptor-β as possible therapeutic targets in desmoid tumors

Background and aim Desmoid tumors are mainly treated by surgical excision and radiotherapy, but the failure to achieve complete response has given rise to the need for investigating the role of possible target therapy. The aim of this study was to evaluate the immunohistochemical detection of cyclooxygenase-2 (COX-2) and estrogen receptor-β (ERβ) in desmoid tumors and to assess their correlation with available clinicopathologic variables. Materials and methods A total of 17 desmoid tumor cases (11 abdominal, five extra-abdominal, and one intra-abdominal) were examined for immunohistochemical detection of COX-2 and ERβ using monoclonal antibodies. Toluidine blue staining was performed to confirm or exclude that COX-2 immunostained cells coincide with mast cells in co-localized sections. Correlation of results with available clinicopathologic variables was done and a P value less than 0.05 was considered significant. Results COX-2 was expressed in tumor cells in 92% of examined desmoid cases (16/17). Toluidine blue staining has shown that COX-2 immunostained cells do not coincide with the few metachromatically stained mast cells in co-localized sections. ERβ was expressed in 67.1% of tumor cells in desmoid cases (11/17); eight cases displayed high ERβ expression and three cases displayed low ERβ expression. No significant correlation was detected between ERβ or COX-2 immunohistochemical expression and patient’s age, sex, tumor size, site, margins status, and recurrence history (P>0.05). Conclusion This study confirmed the immunohistochemical expression of COX-2 and ERβ in tumor cells of the majority of studied desmoid cases. These results introduce COX-2 and ERβ as potential therapeutic targets in desmoid tumors. Further studies with a large sample size and follow-up are recommended to validate the current results.

Dual expression of immunoreactive estrogen receptor β and p53 is a potential predictor of regional lymph node metastasis and postoperative recurrence in endometrial endometrioid carcinoma.

Although histological grade and muscular invasion are related to the malignant behaviors of endometrial endometrioid carcinoma, lymphatic and/or distant metastases are unexpectedly encountered, even in patients in the low-risk group. To re-evaluate additional reliable parameters to predict the risk of progression, we examined the immunohistochemical expression profiles of p53 and estrogen receptor (ER) β proteins. Patients with endometrial endometrioid carcinoma who underwent surgical treatment at our hospital (n = 154) were recruited to this study, and the significance of the relationships between the incidence of regional lymph node metastasis and/or postoperative recurrence and clinical or experimental parameters was evaluated. By multivariate analysis, we found that histological grades, detection of immunoreactive p53 (positive rates more than 10%, p53-stained), and high expression of ERβ (high-ERβ) were independently associated with metastasis and/or recurrence. Among these parameters, the sensitivity and negative predictive values of high-ERβ were very high (up to 100%). In the population with high-ERβ, the positive rates of metastasis and/or recurrence were 61.1% in the p53-stained group and 21.9% in the p53-non-stained (negative) group. Furthermore, the positive rate in the group showing myometrial invasion of more than 1/2 and showing both p53-stained and high-ERβ was 80%. The disease-free survival of patients who were double-positive for p53-stained and high-ERβ was significantly shorter than that in other patients. In summary, our findings showed that increases in ERβ and p53 immunoreactivity were significantly correlated with the incidence of metastasis and/or recurrence in endometrial endometrioid carcinoma, suggesting that double-positivity for p53-stained and high-ERβ may provide a promising clinical indicator to predict the risk of progression.

Estrogen receptor beta expression and prognosis of diffuse large B cell lymphoma

ABSTRACTObjectives: Estrogen receptor beta (ERβ)-selective agonists inhibited B cell lymphoma growth in animal models. However, a recent study found that higher ERβ expression in tissue from diffuse large B cell lymphoma (DLBCL) patients indicated a poorer survival. This study aimed to determine the ERβ expression in DLBCL tissue using immunohistochemistry and correlate with clinical outcomes.Methods: Diagnostic tissues from newly diagnosed adult DLBCL patients treated with Rituximab-Cyclophosphamide/Doxorubicin/Vincristine/Prednisolone were counted for ERβ1-expressing cells. Nodal lymphoma (N = 41) was analyzed separately from extra-nodal DLBCL (N = 31).Results: On immunohistochemistry, ERβ1 was expressed in 73.6% of cases with the median expressing cells of 20%. For nodal lymphoma, high ERβ expression (≥25%) was associated with poorer event free survival (EFS) independent of the international prognostic index with the adjusted hazard ratio (HR) of 2.49 (95% Confidence interval (CI) 1.03–6.00, P = 0.042). On the contrary, high ERβ expression (≥25%) was associated with superior outcomes in extra-nodal DLBCL with the adjusted HR of 0.25 (95% CI 0.09–0.75, P = 0.013) for EFS and adjusted HR of 0.29 (95% CI 0.10–0.85, P = 0.024) for overall survival in multivariate analyses.Conclusion: ERβ1 protein expression represented opposite prognostic factors in nodal vs. extra-nodal DLBCL.

Association of the oestrogen receptor beta with hormone status and prognosis in a cohort of female patients with colorectal cancer

BackgroundThe oestrogen receptor beta (ERβ) is the predominant oestrogen receptor in the normal colon mucosa and has been reported to exert anti-proliferative and pro-apoptotic effects. However, the role of ERβ in colorectal cancer (CRC) progression remains unclear. AimTo investigate the role of ERβ and its association with hormone status and lifestyle indicators in a female cohort of patients with CRC. MethodsTissue microarrays of primary CRC tumour samples from 320 female patients were conducted with a monoclonal anti-ERβ antibody. The staining intensity was evaluated using immunohistochemistry. The association of ERβ expression with overall survival, disease-free survival, hormone status and lifestyle was evaluated, and effect estimators with 95% confidence intervals (CIs) were reported. ResultsAmong the 314 samples with successfully detected ERβ, 182 (58%) had low expression and 132 (42%) had high expression. The Cox multivariate analysis indicated that patients with high ERβ expression had a decreased risk of overall mortality by 50% (hazard ratio [HR], 0.50; CI, 0.30–0.83) and of cancer recurrence by 76% (HR, 0.24; CI, 0.11–0.52) after adjusting for age, tumour-node-metastasis stage and tumour intravascular invasion. Furthermore, high ERβ expression was significantly correlated with shorter breastfeeding time and longer use of hormone replacement therapy. No association was found between ERβ expression and lifestyle indicators. ConclusionElevated ERβ expression is independently associated with a better prognosis and hormone status but not lifestyle indicators in female CRC patients.

Abstract 5640: The prognostic role of estrogen receptor beta in a female cohort of colorectal cancer

Abstract Background: The estrogen receptor beta (ERβ) is the predominant estrogen receptor in the colon mucosa, and has been reported to have anti-proliferative and pro-apoptotic effects. However, the role of estrogen signaling in colorectal cancer (CRC) progression remains unclear. Aim: To investigate the prognostic role of ERβ in a female cohort of CRC. Methods: A tumor tissue microarray of primary CRCs from 333 female patients was stained with a mouse monoclonal anti-ERβ antibody. The intensity of staining was evaluated using immunohistochemistry technique. The expression of ERβ was scored using the semi-quantitative method [IRS (immune-reactive score) = SI (staining intensity) x PP (percentage of positive nuclei)]. Overall survival (OS) and disease-free survival (DFS) were the primary outcomes. Preliminary results are presented. Results: From 321 patients eligible for survival analysis only 315 had successful measurement of ERβ (141 with low expression and 174 with high expression). The univariate analysis showed no effect of ERβ on OS, but high ERβ was associated with lower risk of cancer recurrence as indicated by DFS (HR, 0.61; CI, 0.38-1.02). Cox multivariate analysis indicated that, compared to low expression, high expression of ERβ was significantly associated with a decreased risk of cancer recurrence (HR, 0.39; CI, 0.20-0.77), after adjustment for age, tumor stage, tumor intravascular invasion and hormonal contraception use. Likewise, high ERβ expression was associated with better OS in patients with cancer stage I-III (HR, 0.52; CI, 0.29-0.94) that did not receive adjuvant treatment after operation. Premenopausal women had a higher mean of IRS for ERβ compared to postmenopausal women (p = 0.03). Conclusion: High levels of ERβ significantly correlate with a decreased risk of cancer recurrence in CRC patients. We also noted longer overall survival in the subgroup of patients with cancer stage I-III that had not received adjuvant treatment after operation. The same results of OS were observed for colon cancer patients, but not for rectal cancer patients. A possible explanation could be that most of the rectal cancer patients have undergone radiotherapy before surgery, which might affect the status of ERβ. Citation Format: Geriolda Topi, Roy Ehrnström, Ingrid Palmquist, Marie-Louise Lydrup, Anita Sjölander. The prognostic role of estrogen receptor beta in a female cohort of colorectal cancer [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 5640. doi:10.1158/1538-7445.AM2017-5640