Progesterone Receptor Expression Levels Research Articles

Endocrine Disrupting Effects of Triclosan on the Placenta in Pregnant Rats.

Triclosan (TCS) is a broad-spectrum antimicrobial agent that is frequently used in pharmaceuticals and personal care products. Reports have shown that TCS is a potential endocrine disruptor; however, the potential effects of TCS on placental endocrine function are unclear. The aim of this study was to investigate the endocrine disrupting effects of TCS on the placenta in pregnant rats. Pregnant rats from gestational day (GD) 6 to GD 20 were treated with 0, 30, 100, 300 and 600 mg/kg/d TCS followed by analysis of various biochemical parameters. Of the seven tissues examined, the greatest bioaccumulation of TCS was observed in the placenta. Reduction of gravid uterine weight and the occurrence of abortion were observed in the 600 mg/kg/d TCS-exposed group. Moreover, hormone detection demonstrated that the serum levels of progesterone (P), estradiol (E2), testosterone (T), human chorionic gonadotropin (hCG) and prolactin (PRL) were decreased in groups exposed to higher doses of TCS. Real-time quantitative reverse transcriptase-polymerase chain reaction (Q-RT-PCR) analysis revealed a significant increase in mRNA levels for placental steroid metabolism enzymes, including UDP-glucuronosyltransferase 1A1 (UGT1A1), estrogen sulfotransferase 1E1 (SULT1E1), steroid 5α-reductase 1 (SRD5A1) and steroid 5α-reductase 2 (SRD5A2). Furthermore, the transcriptional expression levels of progesterone receptor (PR), estrogen receptor (ERα) and androgen receptor (AR) were up-regulated. Taken together, these data demonstrated that the placenta was a target tissue of TCS and that TCS induced inhibition of circulating steroid hormone production might be related to the altered expression of hormone metabolism enzyme genes in the placenta. This hormone disruption might subsequently affect fetal development and growth.

Expression of progesterone receptors is significantly impaired in the endometrium of infertile women during the implantation window: a prospective observational study

Objective: To compare the expression of progesterone receptors (A + B) and type-B progesterone receptors in the epithelial and stromal cells of fertile and infertile women.Methods: Women were divided into two groups, the group of fertile controls (group 1) and the group of infertile women (group 2) and were set on regular ultrasound imaging in order to detect ovulation. An endometrial biopsy was obtained on the seventh or eighth post-ovulatory day. Immunohistochemistry was performed to measure percentage of positive nuclei, intensity of staining and h-score for progesterone receptors (PgR) (A + B) as well as type-B progesterone receptors in epithelial and stromal cells. Secondary outcomes included endometrial tissue dating, the rate of tissues being out-of-phase and endometrial thickness.Results: Endometrial issue was obtained from 15 fertile and 30 infertile women. Expression of PgR (A + B) and PgR type-B was significantly lower in the epithelial cells of infertile women. PgR (A + B) h-score was 220.0 ± 18.5 for fertile versus 147.3 ± 18.0 for infertile women (p = 0.02). PgR type-B h-score in epithelial cells was 166.8 ± 30.7 for fertile versus 90.8 ± 20.6 for infertile (p = 0.04). No significant difference was observed in stromal cells.Conclusions: Expression levels of PgR (A + B) as well as type-B receptors are significantly lower in the epithelial cells of infertile women during implantation window.

Correlation of the SUVmax of FDG-PET and ADC values of diffusion-weighted MR imaging with pathologic prognostic factors in breast carcinoma

PurposeTo correlate both primary lesion maximum standardized uptake values (SUVmax) of FDG-PET/CT, and apparent diffusion coefficient (ADC) values of diffusion-weighted imaging (DWI) with clinicopathologic prognostic factors in patients with breast carcinoma. Materials and methods214 patients with 216 mass-type invasive breast carcinomas underwent whole-body FDG-PET/CT and 3-Tesla breast MRI including DWI before initial therapy. The primary tumor’s SUVmax and ADC values were measured using FDG-PET/CT and DWI, respectively. Histologic analysis parameters included tumor size, expression of estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2), and Ki-67, nuclear grade, histology subtype, and axillary lymph node (LN) metastasis. The relationships among SUVmax, ADC values, and pathologic prognostic factors were evaluated. ResultsThe mean SUVmax and ADCmean were 5.63±3.79 (range, 1.2–24.17) and 894±204×10−6mm2/s (range, 452–1550×10−6), respectively. There was a significant but weak inverse correlation between the SUVmax and ADCmean values (correlation coefficient r=−0.30, p<0.0001). SUVmax was associated with numerous prognostic factors such as tumor size (p<0.0001), expression levels of ER (p=0.00041), PR (p=0.00028), HER2 (p=0.00021), and Ki-67 (p<0.0001), nuclear grade (p<0.0001), histology subtype (p=0.00061), axillary LN metastasis (p<0.0001), and TNM staging (p<0.0001). Meanwhile, ADCmean value was associated with tumor size (p=0.013), expression of Ki-67 (p=0.0010), histology subtype (p=0.00013), axillary LN metastasis (p=0.00059), and TNM staging (p=0.0011). ConclusionsPrimary tumor SUVmax on FDG-PET/CT has a stronger relationship with known prognostic parameters and may be a more useful for predicting the prognosis of breast carcinoma than ADC values.

Abstract P6-03-14: Expression of genes for aromatase inhibitor targets to discriminate invasive lobular from invasive ductal carcinomas of the breast using LCM-procured cells to complement endocrine biomarkers

Abstract Background: The BIG 1-98 trial and later the ABCSG 8 study reported that patients with invasive lobular carcinoma (ILC) exhibited better response to aromatase inhibitors (AIs) compared to those with invasive ductal carcinoma (IDC). Aromatase cytochrome P450 (CYP19) synthesizes estrogen from androgens and is the target of AI's. CYP19 substrates are generated by upstream enzymes including estrone sulfatase (SULT1E1) and 3b-hydroxysteroid dehydrogenase type 1 (HSD3B1). Enzymes of the aromatase pathway have been reported to be expressed in intact tissue biopsies of breast cancer. To learn more about the pathogenic mechanisms that may underlie the survival benefit of ILCs treated with AIs, we analyzed expression levels of key enzymes related to the aromatase pathway in ILC and IDC. Unlike previous studies, we determined gene expression levels directly on pure populations of carcinoma cells procured by laser capture microdissection, eliminating the contribution of non-cancerous cells. Methods: Using an IRB-approved biorepository and database, total RNA was extracted from carcinoma cells of 247 de-identified biopsies to perform microarray analyses of 22,000 genes. Of the 247 samples, 16 were ILC, 13 were low grade IDC, 55 were intermediate grade IDC and 85 were high grade IDC, and 107 of these were hormone receptor positive. CYP19, HSD3B1 and SULT1E1 expression was directly detected in LCM-procured breast carcinoma cells of ILC and of IDC. Expression of other genes generally associated with the aromatase pathway, e.g., NADPH-cytochrome P450 reductase (POR), ATP-binding cassette gene (ABCG2), catechol-o-methyltransferase (COMT) and uridine-5'-diphosphate glucuronosyltransferase (UGT1A3 &amp; UGT1A9) as well as HSD17B2 were assessed with LCM-procured cells. Estrogen receptor (ER) and progesterone receptor (PR) proteins were quantified by radio-ligand binding and EIA, and gene expression was validated by qPCR. Results: Univariate Cox regression analyses indicated that ABCG2, HSD17B2and UGTA3 independently predicted disease free and/or overall survival of breast carcinomas. We found that CYP19 expression in carcinoma cells, as well as SULT1E1, COMT, POR, HSD17B2 and UTG1A3 expression, decreased as either ER or PR protein increased. HSD3B1 appeared to be over-expressed in ILC compared to IDC, however this difference did not approach statistical significance, likely due to the small sample size. No differences were seen in expression levels of CYP19 and SULT1E1 between ILC and IDC. Conclusions: An inverse relationship between CYP19 and ER and PR expression levels was observed and suggests that synthesis of estrogens by breast cancer cells in situ plays a significant role in defining tumor biology. Our results also indicate overexpression of HSD3B1 in ILC, although not statistically significant. This finding suggests that HSD3B1 may be a key contributor to the increased benefit of AI therapy seen in ILC. Collectively our results suggest a comprehensive study is warranted to ascertain the molecular basis for differences in expression of genes directing estrogen synthesis in situ in relationship to AI therapeutic responses of histologic subtypes of breast carcinomas. Citation Format: Sanders MAG, Daniels MW, Wittliff JL. Expression of genes for aromatase inhibitor targets to discriminate invasive lobular from invasive ductal carcinomas of the breast using LCM-procured cells to complement endocrine biomarkers. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P6-03-14.

Abstract P5-12-12: Hormone receptor expression level and nuclear grade associated with late recurrence in estrogen receptor-positive breast cancer patients

Abstract Background: It is not known which population of estrogen receptor (ER)-positive breast cancer patients should continue endocrine treatment beyond 5 years to overcome late recurrences. The aim of this study was to examine a combination of nuclear grade (NG) and expression level of ER and progesterone receptor (PR) to predict late recurrences. Methods: We assessed retrospectively 1677 consecutive ER-positive/HER2-negative patients who underwent surgical resection between 2004 and 2009. Patients with T2 or larger tumor and/or node-positive received pre- or postoperative chemotherapy following the international consensus panel from the St Gallen Conference, 2003. All patients had received adjuvant endocrine treatment. NG, ER and PR statuses were determined by immunohistochemistory on surgical specimen. We classified the patients into 3 groups as follows; ER-high (+++ or Allred score 7, 8)/PR-high (++ to +++ or 5-8) (n=212), ER-high / PR-low (- to + or 3-6) (n=208), and ER-low (+ to ++ or 3-6) / PR-any (n=255). We compared distant disease-free survival (DDFS) in each cohort based on the NG (1:low, and 2 or 3: high). Results: A median follow-up period was 77.0 months. Four hundred sixty seven patients (27.8%) received neoadjuvant chemotherapy, 208 patients (12.4%) received adjuvant chemotherapy, and 1002 patients (59.8%) did not received chemotherapy. Of the 467 patients with neoadjuvant chemotherapy, 65 patients (13.9%) had developed distant metastasis during study period (before 5 years in 51 (11.0%); and after 5 years in 14 (3.0%)). NG-low had significantly higher risk of late recurrence after 5 years than that of NG-high (p=0.005). According to hormonal receptor expression levels, in patients with NG-low, ER-low/PR-any had significantly higher overall DDFS rate than ER-high/PR-low (p=0.016). A similar trend was found before 5 years (p=0.077). However, ER-high/PR-high turned to have significantly high risk of recurrence after 5 years compared to ER-low/PR-any (p=0.024). Of the 208 patients with adjuvant chemotherapy, 16 patients (7.6%) had developed distant metastasis during study period (before 5 years in 5 (2.4%), and after 5 years in 11 (5.2%)). there was no association between a risk of recurrence and hormone receptor statuses at any study period. In the patients did not received chemotherapy, ER-high/PR-high had a trend of higher DDFS rate than others before 5 years (p=0.067). Of the 1002 patients without chemotherapy, only 36 patients (3.6%) had developed distant metastasis during study period (before 5 years in 27 (2.7%), and after 5 years in 9 (0.9%)). There was no difference of late recurrence after 5 years among the patients regardless of ER and PR expression level and NG with only low recurrence rate (0.9%). Conclusions: Our results demonstrated that, in ER-positive/HER2-negative patients who underwent neoadjuvant chemotherapy, NG-low/ER-high/PR-high should receive extend hormonal treatment over 5 years because of the high risk of late recurrence but NG-high/ER-high might not need. Furthermore, patients with T1 and node-negative may not need extend hormonal treatment because of the extremely low risk of late recurrence regardless of NG and hormone receptor statuses. Citation Format: Iwase M, Hayashi N, Yoshida A, Kajiura Y, Takahashi Y, Takei J, Suzuki K, Nakamura S, Yamauchi H. Hormone receptor expression level and nuclear grade associated with late recurrence in estrogen receptor-positive breast cancer patients. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P5-12-12.

Immunohistochemical Performance of Estrogen and Progesterone Receptor Antibodies on the Dako Omnis Staining Platform: Evaluation in Multicenter Studies.

The analysis of estrogen receptor (ER) and progesterone receptor (PR) expression levels by immunohistochemistry is an important part of the initial evaluation of breast cancer and critically important in treatment planning. Anti-ERα (clone EP1) and anti-PR (clone PgR 1294) antibodies are in development for the Dako Omnis automated staining platform. These antibodies are not yet commercially available and are in performance evaluation, including the 4 international, multicenter studies reported here. For each antibody, a reproducibility study and a method comparison study was done in a randomized manner in order to test the antibodies under conditions closest to real-world user conditions. The reproducibility studies included 5 staining runs on the Dako Omnis with 20 formalin-fixed and paraffin-embedded human breast carcinoma specimens in 3 independent laboratories, and the method comparison studies included several hundred specimens stained on the Dako Omnis and on the Autostainer Link 48 platforms. Stained slides were evaluated for nuclear ER or PR expression according to American Society of Clinical Oncology/College of American Pathologists guidelines (≥1% cut-off for positive) by pathologists who were blinded from the staining method and specimen ID. For both anti-ERα (clone EP1) and anti-PR (clone PgR 1294) on the Dako Omnis, high reproducibility agreement rates were obtained on the interrun, interlaboratory, and interobserver endpoints. High concordance rates were observed between the specimens stained on the Dako Omnis platform and the Autostainer Link 48 platform. Staining quality was excellent for both anti-ERα (clone EP1) and anti-PR (clone PgR 1294) on the Dako Omnis. These results suggest that these antibodies are reliable and reproducible tools for immunohistochemistry analysis of ER and PR expression levels in formalin-fixed and paraffin-embedded breast carcinoma tissues on the Dako Omnis platform.

Analysis of factors affecting endocrine therapy resistance in breast cancer.

The present study aimed to identify the factors involved in the resistance to endocrine therapy in breast cancer (BC) patients with a positive estrogen receptor status via the collection of clinical, pathological and immunohistochemical indices. A retrospective survey was performed in patients who experienced the relapse and metastasis of BC between November 2007 and March 2013. A total of 45 patients were enrolled, and the observational duration was 7-84 months. The Kaplan-Meier method was used to create a survival curve, while the log-rank test was used to analyze the survival curve and the Cox regression analysis was used to investigate the associated factors contributing to the resistance to endocrine therapy. Univariate analysis showed that the age of onset, the use of radiotherapy, the endocrine treatment program, and the expression levels of progesterone receptor (PR) and CerbB2 affected the impact of endocrine treatment. The Cox regression analysis indicated that the age of onset, the use of radiotherapy, and the expression levels of PR and CerbB2 affected the disease-free survival time after endocrine therapy. A young age of onset, not receiving radiotherapy, a low expression level of PR and a high expression level of CerbB2 were the risk factors involved in the resistance to endocrine therapy in patients with BC.

A75: Modern concepts of cellular and molecular mechanisms of breast cancer and their prognostic significance

Breast cancer is one of the most common cancers in women. The incidence of breast cancer in 2014 in the Republic of Mordovia was 69.9 per 100,000 female populations. Breast cancer occurs when excessive expression of oncoproteins switches in the case of transformation of proto-oncogene in PRADI. In primary breast tumors, mutations and the expression of the three oncogenes Her2/neu, C-mys, Int-2, as well as in supressonyh genes – the p53 gene and the retinoblastoma gene RB are the most common. Several studies found that oncogene – C-mys was expressed in 16.8% of primary breast cancer cases and in 35% cases with subsequent development of metastases. Proteins that stimulate the phosphorylation of mitogen-activated protein kinasescan activateunder the influence of growth factors. Development of breast cancer is regulated by a complex interaction of many hormones and growth factors. Currently, one of the leading theories of developing breast cancer is the increased hormonal stimulation of proliferative processes in the development of neoplasia. One of the manifestations of hormonal imbalance in tumor during a regular decrease in blood competitive inhibitor of the biological effects of estrogen – progesterone, which is in correlation with the stage of the spread of neoplasia. The role of the overproduction of estrogen in the pathogenesis of breast cancer is confirmed by the fact that in women who underwent oophorectomy before the age of 38 years, the risk of breast cancer development is 1.5 times less than in those who did not have such operation. Excessive accumulation of lipid peroxidation products in the area of neoplasia activates mechanisms violation of intercellular interaction that caused the destruction of the lipid components of membranes. The study group included 112 patients treated at the State Institution of Health of the Republic of Mordovia “National Oncology Center”. To identify the nature of tumors, all patients underwent immunohistochemical analysis. Androgen receptors were found in 48% of breast cancer cases, the expression level of androgen receptor in the tumor was much lower than the expression level of estrogen and progesterone receptors. Low levels of progesterone receptor expression in breast cancer cells were combined with high levels of expression of Ki-67 antigen, HER-2 oncoprotein in tumor cells. Patients with HER-2 (3+) and (2+) had more frequent multiple metastases in lymph nodes compared to patients with HER-2 (0) and (1+) phenotypes. Maximum expression of HER-2 oncoprotein in tumor cells indicated high metastatic potential and poor prognosis. It may be concluded that the cellular and molecular mechanisms of breast cancer are complex. Therefore, carcinogenesis has a “multistep” nature and at least two or more mutations in the cells of the same clone – parent and child are required to generate malignant tumors. Thus, the development of oncogenic transformation does not necessarily mean the process of tumor formation.

Oncotype DX in Bilateral Synchronous Primary Invasive Breast Cancer.

Synchronous bilateral breast cancers frequently share the same estrogen receptor (ER) status, yet may differ in other histopathologic features. We sought to examine concordance rates of Oncotype DX recurrence score (RS) testing in women with synchronous bilateral ER-positive breast cancer. Institutional databases were reviewed to identify patients with synchronous (within 6 months) bilateral primary invasive breast cancer and multiple RSs. RSs were stratified by risk group (RS < 18, low; RS 18-30, intermediate; RS ≥ 31, high) and considered discordant if they reflected different risk groups. From 2005-2014, a total of 115 patients presented with synchronous bilateral invasive breast cancer; 43 (37 %) had two RSs available. Median patient age was 60 years (42-84), median tumor size was 1.2 cm (0.5-3.7), and all cases were HER2 negative and node negative. Of 86 RSs, 63 (73 %) were low risk, 20 (23 %) were intermediate risk, and 3 (3 %) were high risk. RSs were concordant in 29 (67 %) patients. Patients with concordant RSs were older (62 years vs. 56 years) and had median levels of progesterone receptor (PR) expression that were higher and more similar-80 and 85 % for bilateral cancers, respectively, compared with 55 and 75 % for bilateral cancers in discordant cases. Discordant RS led to a treatment change in 8/14 (57 %) cases. Among women with synchronous bilateral ER-positive HER2-negative breast cancer, RSs were concordant in 67 % of cases. Concordance rates may be higher in older women or among those with comparable levels of PR expression. These data suggest that testing of both tumors should be considered in patients who are candidates for adjuvant chemotherapy.

Impact of biomarker changes during neoadjuvant chemotherapy for clinical response in patients with residual breast cancers.

Residual cancer burden or Ki67 expression levels in residual tumors reportedly provided significant prognostic information for a non-pathological complete response subset after neoadjuvant chemotherapy (NAC). However, the significance of Ki67 reduction for clinical response during chemotherapy in each subtype or menopausal status is yet to be determined. A total of 183 breast cancers surgically removed after chemotherapy were recruited for this study. Expression levels of estrogen receptor (ER), progesterone receptor (PgR), and Ki67 were determined immunohistochemically for semiquantitative measurement and these biomarkers were compared in pre- and post-NAC samples from pathological non-responders (n = 125). Responses to chemotherapy were evaluated both clinically and pathologically. Ki67 expression levels after NAC (median 5 %, range 0-70 %) were significantly reduced compared with before NAC (25, 1-80 %, P < 0.0001), but only in patients who attained clinical response. This significant suppression of Ki67 in clinical responders was consistently observed in breast cancers from the ER-positive subset, but not the ER-negative subset in the total test set (n = 120). These observations were also made in the validation set (n = 63). Among premenopausal, but not postmenopausal patients, a significant decrease in PgR expression levels was detected in breast cancers of patients who attained clinical response (pre-NAC 50, 0-100 %, post-NAC 5, 0-20 %; P = 0.0003). The impact of Ki67 suppression on clinical response seems to be restricted to ER-positive breast cancers. Since PgR expression levels of premenopausal ER-positive cancers were significantly reduced in clinical responders, inhibition of estrogen signaling due to chemotherapy-induced amenorrhea may be involved in this association.

Expression of the estrogen receptor α, progesterone receptor and epidermal growth factor receptor in papillary thyroid carcinoma tissues.

The present study aimed to determine the protein expression, in addition to the clinical value of the expression, of estrogen receptor α (ERα), progesterone receptor (PR) and epidermal growth factor receptor (EGFR) in papillary thyroid carcinoma (PTC). The expression of ERα, PR and EGFR was examined immunohistochemically on paraffin-embedded thyroid tissues obtained from 64 patients with PTC and 14 patients with nodular thyroid goiter (NTG). The expression level of ERα, PR and EGFR was found to be significantly elevated in the PTC tissues compared with the NTG tissues. In addition, the expression of ERα was found to be correlated with the size of PTC tumors. However, there was no significant difference between the expression levels of ERα, PR and EGFR in males and females with PTC. Thus, immunohistochemical evaluation of ERα, PR and EGFR expression in patients with PTC may aid in the prediction of the prognosis of patients with PTC.

Divergent behavior of cyclin E and its low molecular weight isoforms to progesterone-induced growth inhibition in MCF-7 cells.

Background:Progesterone is a steroid hormone that modulates proliferation and differentiation in a cell phase and tissue-specific manner. Its function in breast cancer cells is of great significance since it can predict susceptibility of tumor cells to inhibitory effects of progesterone as adjuvant therapy.Materials and Methods:Stable clones overexpressing cyclin E (EL) and its low molecular weight isoforms (LMW-Es) were generated and treated with various concentrations of progesterone. Cell proliferation was assessed 24 and 48 h after the treatment. Changes in progesterone receptor (PR) expression were measured by real-time polymerase chain reaction.Results:Here we demonstrated that overexpression of EL and LMW-Es have divergent effects with regard to progesterone response. We found that progesterone could significantly decrease the growth rate of EL-expressing cells in the second cell cycle after treatment; however, progesterone was ineffective to arrest growth of LMW-Es expressing cells. PR expression level was at control level in EL-expressing cells but was downregulatedin LMW-Esexpressing clones.Conclusion:These results were in line with progesterone response of studied cells. The drop in PR expression together with altered distribution of p21 and p27 can explain different effects of cyclin E isoforms expression on progesterone responsivity. These data bring cyclin E status of cancer cells as a marker for predicting the efficacy of progesterone treatment.

Serum levels of sex hormones and expression of their receptors in thyroid tissue in female patients with various types of thyroid neoplasms

Previous studies have demonstrated the expression of estrogen receptor (ER) and progesterone receptor (PR) in thyroid cancer; however, little is known regarding the levels of estrogen, progesterone, follicle-stimulating hormone (FSH), and luteinizing hormone (LH) in serum and the expression of ER, PR, FSH receptor (FSHR), and LH receptor (LHR) in thyroid tissues of patients with different types of thyroid neoplasms. Serum levels of estrogen, progesterone, FSH, and LH were measured by chemiluminescence, and expression of ER, PR, FSHR, and LHR in thyroid tissue was detected by immunohistochemistry in female patients with thyroid adenoma (n=70), nodular goiter (n=73), thyroid papillary cancer (n=149), poorly differentiated thyroid carcinoma (n=12), or undifferentiated thyroid carcinoma (n=8) and in normal controls (n=60). The positive rates of serum estrogen level and ERα expression were significantly greater in patients with various types of thyroid neoplasms than in normal controls. The positive rates of ERβ expression were significantly less in various types of thyroid neoplasms than in normal thyroid tissues, especially in poorly differentiated carcinoma and undifferentiated carcinoma. The negative rates of serum progesterone level and positive rates of PR expression in thyroid tissue were significantly greater in patients with thyroid adenoma, nodular goiter, or thyroid papillary cancer than in normal controls. The positive rates of serum FSH and LH levels and FSHR and LHR expression were significantly greater in the thyroid adenoma group than in other groups. Our findings suggest that thyroid neoplasms might be sex hormone-dependent. The positive expression of ERα and PR often indicates thyroid papillary carcinoma, and the ERβ expression status is important for the diagnosis of poorly differentiated carcinoma and undifferentiated carcinoma. In addition, thyroid adenoma is often accompanied by an increase in serum FSH and LH levels, as well as FSHR and LHR expression. Thus, the combined detection of serum levels of sex hormones and expression of their receptors allows for a differential diagnosis and evaluation of the degree of differentiation among various types of thyroid neoplasms.

Selective Estrogen Receptor Modulation by Larrea nitida on MCF-7 Cell Proliferation and Immature Rat Uterus.

Larrea nitida is a plant that belongs to the Zygophyllaceae family and is widely used in South America to treat inflammatory diseases, tumors and menstrual pain. However, its pharmacological activity remains unclear. In this study we evaluated the property of selective estrogen receptor modulator (SERM) of Larrea nitida extracts (LNE) as a phytoestrogen that can mimic, modulate or disrupt the actions of endogenous estrogens, depending on the tissue and relative amount of other SERMs. To investigate the property of SERM of LNE, we performed MCF-7 cell proliferation assays, estrogen response element (ERE)-luciferase reporter gene assay, human estrogen receptor (hER) binding assays and in vivo uterotrophic assay. To gain insight into the active principles, we performed a bioassay-guided analysis of LNE employing solvents of various polarities and using classical column chromatography, which yielded 16 fractions (LNs). LNE showed high binding affinities for hERα and hERβ with IC50 values of 1.20 ×10−7 g/ml and 1.00×10−7 g/ml, respectively. LNE induced 17β-estradiol (E2)-induced MCF-7 cell proliferation, however, it reduced the proliferation in the presence of E2. Furthermore, LNE had an atrophic effect in the uterus of immature rats through reducing the expression level of progesterone receptor (PR) proteins. LN08 and LN10 had more potent affinities for binding on hER α and β than other fractions. Our results indicate that LNE had higher binding affinities for hERβ than hERα, and showed SERM properties in MCF-7 breast cancer cells and the rat uterus. LNE may be useful for the treatment of estrogen-related conditions, such as female cancers and menopause.

Disruption of the hormonal network and the enantioselectivity of bifenthrin in trophoblast: maternal-fetal health risk of chiral pesticides.

Endocrine-disrupting chemicals (EDCs) can interfere with normal hormone signaling to increase health risks to the maternal-fetal system, yet few studies have been conducted on the currently used chiral EDCs. This work tested the hypothesis that pyrethroids could enantioselectively interfere with trophoblast cells. Cell viability, hormone secretion, and steroidogenesis gene expression of a widely used pyrethroid, bifenthrin (BF), were evaluated in vitro, and the interactions of BF enantiomers with estrogen receptor (ER) were predicted. At low or noncytotoxic concentrations, both progesterone and human chorionic gonadotropin secretion were induced. The expression levels of progesterone receptor and human leukocyte antigen G genes were significantly stimulated. The key regulators of the hormonal cascade, GnRH type-I and its receptor, were both upregulated. The expression levels of selected steroidogenic genes were also significantly altered. Moreover, a consistent enantioselective interference of hormone signaling was observed, and S-BF had greater effects than R-BF. Using molecular docking, the enantioselective endocrine disruption of BF was predicted to be partially due to enantiospecific ER binding affinity. Thus, BF could act through ER to enantioselectively disturb the hormonal network in trophoblast cells. These converging results suggest that the currently used chiral pesticides are of significant concern with respect to maternal-fetal health.

Proposed new clinicopathological surrogate definitions of luminal A and luminal B (HER2-negative) intrinsic breast cancer subtypes.

IntroductionThe St Gallen International Expert Consensus on the Primary Therapy of Early Breast Cancer 2013 recognized substantial progress in the pathological characterization of breast cancer subtypes. A useful surrogate definition was developed to distinguish luminal A–like breast cancer from luminal B–like disease based on a combination of estrogen receptor (ER), progesterone receptor (PgR) and Ki-67 status, without a requirement for molecular diagnostics. Differences depend upon the choice of the threshold value for Ki-67 and the requirement for substantial PgR positivity. We aimed to verify the suitability of the new surrogate definitions of luminal subtypes in terms of distant disease control in a large series of patients.MethodsWe studied 9,415 women with a median follow-up of 8.1 years who (1) had ER-positive, human epidermal growth factor receptor 2 (HER2)–negative early breast cancer and (2) had undergone surgery at the European Institute of Oncology between 1994 and 2006. We evaluated distant disease-free survival of patients with “low” (<14%), “intermediate” (14% to 19%) or “high” (≥20%) Ki-67 positivity stratified by PgR expression (negative or low versus high). We calculated the cumulative incidence of distant events, considered competing events and performed multivariable analysis adjusted for pathologic tumor stage, pathologic node stage, tumor grade, peritumoral vascular invasion and menopausal status.ResultsLack of substantial PgR positivity was associated with poorer outcomes only for patients with an intermediate Ki-67 level (P < 0.001). The 4,890 patients (51.9%) with low Ki-67 level (any PgR expression level) or with intermediate Ki-67 level but substantial PgR positivity had comparably good outcomes and thus may represent a most advantageous grouping of those with luminal A–like disease.ConclusionsThe updated pathological definition of intrinsic molecular subtypes may maximize the number of patients classified as having the luminal A–like intrinsic subtype of breast cancer and for whom the use of cytotoxic drugs could mostly be avoided.

Prostate stromal cells express the progesterone receptor to control cancer cell mobility.

BackgroundReciprocal interactions between epithelium and stroma play vital roles for prostate cancer development and progression. Enhanced secretions of cytokines and growth factors by cancer associated fibroblasts in prostate tumors create a favorable microenvironment for cancer cells to grow and metastasize. Our previous work showed that the progesterone receptor (PR) was expressed specifically in prostate stromal fibroblasts and smooth muscle cells. However, the expression levels of PR and its impact to tumor microenvironment in prostate tumors are poorly understood.MethodsImmunohistochemistry assays are applied to human prostate tissue biopsies. Cell migration, invasion and proliferation assays are performed using human prostate cells. Real-time PCR and ELISA are applied to measure gene expression at molecular levels.ResultsImmunohistochemistry assays showed that PR protein levels were decreased in cancer associated stroma when compared with paired normal prostate stroma. Using in vitro prostate stromal cell models, we showed that conditioned media collected from PR positive stromal cells inhibited prostate cancer cell migration and invasion, but had minor suppressive impacts on cancer cell proliferation. PR suppressed the secretion of stromal derived factor-1 (SDF-1) and interlukin-6 (IL-6) by stromal cells independent to PR ligands. Blocking PR expression by siRNA or supplementation of exogenous SDF-1 or IL-6 to conditioned media from PR positive stromal cells counteracted the inhibitory effects of PR to cancer cell migration and invasion.ConclusionsDecreased expression of the PR in cancer associated stroma may contribute to the elevated SDF-1 and IL-6 levels in prostate tumors and enhance prostate tumor progression.

Differential expression of progesterone receptor, FOXA1, GATA3, and p53 between pre- and postmenopausal women with estrogen receptor-positive breast cancer

Estrogen receptor (ER) is essential for estrogen-dependent growth, and its level of expression is considered a crucial determinant of response to endocrine therapy and prognosis in ER-positive breast cancer. On the other hand, the clinical role of progesterone receptor (PgR) in ER-positive breast cancer remains controversial, although testing of PgR by immunohistochemistry (IHC) has become routine. Recent studies indicated that plasma estradiol levels were related to the expression levels of estrogen-responsive genes in ER-positive breast cancer tissues in both pre- and postmenopausal women. In this study, we analyzed the expression levels of estrogen-responsive genes (PgR and TFF1), a progesterone-responsive gene (RANKL), ER-related genes (FOXA1 and GATA3), HER2, Ki67 and p53 in ER-positive, HER2-negative breast cancer tissues by IHC. Correlations between the expression levels of these molecular markers and clinicopathological factors, including prognosis, were compared between pre- and postmenopausal women. Serum levels of estrone, estradiol, progesterone, and testosterone were also measured. Expression levels of PgR, TFF1, RANKL, and GATA3 were significantly higher in premenopausal women than in postmenopausal women. Serum estradiol levels were positively correlated with Ki67 labeling index (LI) in premenopausal women, but not in postmenopausal women. High expression of FOXA1 and GATA3 was significantly associated with improved disease-free survival in premenopausal women, but not in postmenopausal women, whereas high expression of PgR and low expression of p53 were significantly correlated with the improved disease-free survival in postmenopausal women, but not in premenopausal women. Moreover, the best cutoff points of Ki67 LI for disease-free survival were 30 % for premenopausal women and 14 % for postmenopausal women. Expression levels of ER, TFF1, and RANKL were not associated with the disease-free survival in either pre- or postmenopausal women. Our results suggest that the mechanisms of development and estrogen-dependent growth of ER-positive breast cancer might differ according to menopausal status.

Abstract P5-05-04: Relationship between plasma estradiol levels and estrogen-regulated gene expressions in premenopausal estrogen receptor-positive breast cancers

Abstract Background: Estrogen receptor (ER) and progesterone receptor (PgR) expression status in breast cancer are important factors in determining the use of adjuvant therapy. Plasma estradiol (E2) and progesterone vary markedly through the menstrual cycle. Recently, the levels of PgR expression were reported to be correlated with E2 at the time of surgery in Caucasian premenopausal breast cancer patients. Materials and methods: In this study, plasma E2 levels at surgery were examined for correlations with expressions of estrogen-regulated genes (ERGs), including PgR, and proliferation genes in 147 Japanese premenopausal ER-positive breast cancers. Plasma E2 and progesterone, FSH, and LH were measured on the day of surgery by ECLIA and mRNA expressions were analyzed for 4 ERGs (PgR, GREB1, TFF1, PDZK1), 2 progesterone-regulated (RANKL, cyclin D1) and 2 proliferation genes (MKI67, BIRC5) by RT-PCR using the TaqMan system. ER and PgR protein expression levels were estimated by immunohistochemistry. Results: Our results revealed that expressions of PgR mRNA and protein in breast cancer tissues were significantly correlated with plasma E2 levels at surgery (p = 0.005, p = 0.0165, respectively). Other ERGs were also significantly correlated with plasma E2 levels. However, there were no correlations between mRNA expressions of proliferation genes, MKI67 and BIRC5, and plasma E2 levels. Furthermore, mRNA expressions of the progesterone-regulated genes, RANKL and cyclin D1, were significantly correlated with PgR mRNA expression (p&amp;lt;0.0001, p&amp;lt;0.0001, respectively). Discussion: Significant correlations were seen in the expression of ERGs in Japanese ER-positive breast cancers according to the plasma E2 levels, which vary markedly through the menstrual cycle. Our results supported the previous report in Caucasian patients by other group. This variability may affect the interpretation of gene expression profiles incorporating ERGs such as the 21-gene recurrence score. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P5-05-04.

Abstract P4-11-01: Effect of letrozole is superior to tamoxifen in limiting preneoplasia in both ERa and aromatase over-expressing mice

Abstract Background: Breast cancer develops as genetic changes give rise to preneoplastic lesions, which may or may not progress to invasive breast cancer. Studies suggest that estrogens may promote this process. Targeted over-expression of both estrogen receptor (Esr1) and aromatase (CYP19A1) to mammary epithelial cells (MECs) of genetically engineered mice results in the development of mammary hyperplasia and invasive cancer. Both of these genetic interventions lead to altered estrogen signaling pathways with increased expression levels of progesterone receptor (PGR) and cyclin D1. Tamoxifen is currently the only FDA-approved drug for chemoprevention of invasive breast cancer in premenopausal high-risk women but while effective it does not absolutely prevent cancer development. Purpose: Test the impact of letrozole and tamoxifen in both tet-op-CYP19A1/MMTV-rtTA mice and tet-op-Esr1/MMTV-rtTA. Methods: Female tet-op-CYP19A1/MMTV-rtTA and tet-op-Esr1/MMTV-rtTA mice were implanted subcutaneously with a pellet containing either letrozole 2.5mg (60-day release), tamoxifen 25mg (60-day release), or placebo at age 10m. Mice were necropsied at age 12m and mammary tissue collected for morphological, histological, and protein expression studies. Whole mounts and hematoxylin and eosin stained slides were used to evaluate growth abnormalities, hyperplastic alveolar nodules (HANs) or cancer. Immunohistochemistry and western blot analyses were used to compare and quantitate changes in protein expression. Results: Letrozole treatment resulted in no HAN detection in both Esr1 and CYP19A1 over-expressing mice. Tamoxifen reduced HAN prevalence to 14 (compared to 60%) in CYP19A1 over-expressing mice; HAN prevalence persisted in 56% (as compared to 67%) in Esr1 over-expressing mice. The reduction in preneoplasia prevalence in letrozole treated mice was accompanied by a statistically significant reduction in the proliferative index in both CYP19A1 and Esr1 over-expressing mice. However, reductions found following tamoxifen exposure were not statistically significant. Letrozole significantly reduced the percentage of ERα positive nuclei and the modified Allred score for cyclin D1 in CYP19A1 over-expressing mice, but not in Esr1 over-expressing mice as compared to control mice. In Esr1 over-expressing mice tamoxifen exposure was associated with a significant increase in the percentage of ERα positive nuclei that was not seen in the CYP19A1 over-expressing mice. PGR expression differences were not statistically significant for either treated group. Conclusions: The impact of letrozole was greater than tamoxifen in reducing prevalence of preneoplasia and MEC proliferation rates in both ERα and aromatase over-expressing mice. These results suggest that letrozole should be considered as a chemopreventive agent based on individual estrogen signaling prognostic factors. Support: P30CA051008, RO1CA112176, T32CA009686-15, KG080359. Support: P30CA051008, RO1CA112176, T32CA009686-15, KG080359. Content solely responsibility of authors and does not necessarily represent official NCI/NIH views. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-11-01.