ERα mRNA Expression Research Articles

Oxytocin effects on the inhibition of the NF-κB/miR195 pathway in mice breast cancer

Oxytocin (OT) has the suppressive effects on breast tumor formation and development. We hypothesized that OT through the NF-κB inhibition can induce the miR-195 up-regulation which it can promote the cell apoptosis and inhibit the cell proliferation. Thirty-two BALB/c female mice were equally divided into four groups to study the effects of OT and atosiban (ATO) (an oxytocin receptor antagonist) on the mammary tumor growth. The animal weight, OT plasma concentration, and the tumor weight and volume were measured. Moreover, the tumor-related signaling pathways including NF-κB, miR-195, and Cyclin D1 were evaluated by qPCR assays, and Akt and ERK proteins were assessed by western blot at the end of the study. The volume and weight of tumors were significantly decreased after OT administration. The phosphorylated Akt and ERK expressions were significantly decreased in the OT group compared to the tumor group. In contrast, the dephosphorylated Akt and ERK expressions were significantly increased in the OT group in comparison with the tumor group. The mRNA expressions of miR-195, OTR, and Bax genes were significantly increased, and the mRNA expression of ERα, PI3K, NF-κB, cyclin D1 and Bcl-2 genes were decreased in the OT group in comparison with the tumor group. Interestingly, ATO administration reversed these effects. These results can exhibit a new therapeutic potential for OT on the down-regulation of the NF-κB and up-regulation of miR-195 and consequently, decrease of the tumor volume and weight in a mouse model of breast cancer.

Effects of calcium and estrogen on the development of the ceratohyal cartilage in zebrafish (Danio rerio) larvae upon embryo and maternal cadmium exposure

The present study is to investigate the reason why the ceratohyal cartilage (CH) angle of zebrafish larvae were larger compared to the control group after their female parents were treated with cadmium (F-Cd). However, the CH angle was smaller compared to the control group when embryos were directly exposed to Cd2+ for 72 h (D-Cd). Results showed that calcium contents of larvae were lower than the control, but the transporter isoforms trpv4 and trpv6 mRNA expressions were significantly increased upon D-Cd treatment. Furthermore, external Ca2+ added during D-Cd treatment reveals that the CH angles of larvae did not appear significantly different compared to the control. On the other hand, E2 (17β-estradiol) contents were higher around 1.9 folds in the ovaries of females; CH angle were over 25°, and Cd2+ contents were higher around 6 folds than the control group on larvae treated through F-Cd treatment; CH angles and E2 levels on larvae were higher than the control after the larvae were treated with 1.84 μM E2 (D-E2); Estradiol receptor (ER) isoforms ERβ1 and ERα mRNA expressions significantly increased when 0 hpf embryos were either treated with D-E2 or D-Cd. According to the results, we suggested that the CH angle of larvae become larger upon F-Cd treatment due to maternal Cd2+ inducing E2 levels. However, the CH angle of larvae appeared to be smaller compared to the control upon D-Cd treatment. We suggested that the CH angle decreased due to the decrease of Ca2+ contents upon Cd2+ exposure.

PO-345 The role of the deubiquitinase USP11 in endocrine-driven breast cancer

IntroductionApproximately 80% of breast cancers overexpress the oestrogen receptor α (ERα) and depend on this key transcriptional regulator for growth. The discovery of novel mechanisms controlling ERα function represent major advances in our understanding of breast cancer progression and potentially offer new therapeutic opportunities. Here, we investigated the role of deubiquitinating enzymes (DUBs), which remove ubiquitin moieties from proteins, in regulating ERα activity in breast cancer.Material and methodsTo identify DUBs involved in ERα transcriptional activity, we performed an RNAi loss-of-function screen using a library of shRNA vectors targeting all 108 known or putative human DUB genes. We found that suppression of the BRCA-associated DUB, USP11, downregulated ERα transcriptional activity in ZR-75–1 cells. Dual luciferase reporter assays and qRT-PCR were used to determine ERα activity in stable ZR-75–1 USP11 knockdown cell lines. PTMScan technology, which allows for enrichment of ubiquitinated proteins and quantitative profiling by mass spectrometry, was used to reveal the ubiquitinome in these cells and results are pending. RNA sequencing (RNA-seq) technology was carried out in estrogen-independent LCC1 USP11 knockdown cells, were USP11 was hypothesised to play a vital role. Finally, immunohistochemistry (IHC) was used to investigate the prognostic relevance of USP11 in breast cancer patients.Results and discussionsKnockdown of USP11 in ZR-75–1 cells decreased ERα transcriptional activity and mRNA expression of ERα target genes. These results were further validated in a HEK293T USP11 CRISPR knockout model with ectopic ERα expression. Interestingly, USP11 expression was upregulated in LCC1 cells, an isogenic, estrogen-independent model derived from MCF-7 cells. Knockdown of USP11 in LCC1 cells decreased the expression of multiple ERα target genes and cell cycle-associated genes, as determined by RNA-seq. IHC staining of a breast cancer tissue microarray (103 ER +patients) and subsequent Kaplan-Meier analysis of this cohort revealed a significant association between high USP11 expression and poor overall (p=0.030) and breast cancer-specific survival (p=0.041). In silico analysis of publically available breast cancer gene expression datasets further supported an association between high USP11 expression and poor prognosis.ConclusionThese results suggest a role for USP11 in ERα transcriptional activity and reveal a novel mechanism as to how this receptor is regulated in breast cancer.

The effect of TGF-β signaling on regulating proliferation of uterine leiomyoma cell via ERα signaling activated by bisphenol A, octylphenol and nonylphenol in vitro.

To study the transforming growth factor beta (TGF-β) signaling pathway in interactions with estrogen receptor alpha (ERα) signaling pathway mediating the growth of human uterine leiomyoma (UL) activated by phenolic environmental estrogens (EEs). The subcultured UL cells were used to determine the validation of TGF-β3 for the viability of human UL cells using CCK-8 assay, mRNA expressions of ERα, and c-fos by quantitative reverse transcription polymerase chain reaction method, and expressions of p-Smad3, SnoN, and c-fos proteins by Western blot assay in each treatment group. Compared with each of EEs or TGF-β3 treatment, slightly decrease in the proliferation rate of UL was detected in the coexistence of each EE with TGF-β3. Interestingly, mRNA expressions of ERα and c-fos reduced in the setting of coexistence of TGF-β3 and EEs. Somehow, the expression of p-Smad3 and c-fos proteins significantly decreased in each of E2, bisphenol A (BPA), nonylphenol (NP), and octylphenol (OP) group, as well as the expression of SnoN protein significantly reduced only in BPA and NP groups, followed by TGF-β3 treatment. With the overlaid action of ICI 182,780, the expression of p-Smad3 protein significantly increased in OP group, but slightly increased in E2, BPA, NP, and OP groups. However, compared with the control group, the expression of SnoN and c-fos proteins significantly decreased in the same setting. Both ERα signaling pathway and TGF-β signaling pathway have different roles in governing UL cell proliferation. The phenolic EEs can be a promoter to the proliferation of UL cells, which is mediated by ERα signaling pathway and cross-talked with TGF-β signaling pathway.

Anti-hyperplasia Effects of Total Saponins From Phytolaccae Radix in Rats With Mammary Gland Hyperplasia via Inhibition of Proliferation and Induction of Apoptosis.

Mammary gland hyperplasia (MGH) is a pathological condition that affects the majority of women at the child-bearing stage. The hormone and endocrinal therapy are typically used to treat MGH. Nevertheless, there are still some certain side effects accompanied with the benefits, which negatively affect the life quality of patients. Therefore, plant-derived agents that are effective against MGH development and with fewer side effects should be developed. The aim of this study was to investigate the protective effects and underlying mechanism of total saponins of Phytolaccae (TSP) against MGH in vivo. The results showed that treatment with TSP could significantly correct the disorder of serum sex hormones levels in rats with MGH, and eliminate the formation of MGH. Moreover, TSP significantly protected estrogen and progesterone-induced MGH histological changes, inhibited the swelling of the nipple, and improved the organ coefficient of uterus in rats with MGH. Mechanistically, TSP treatment not only effectively suppressed the mRNA and protein expression of ERα and PR in mammary gland, but also simultaneously up-regulated ERβ expression, and thus blocked sex hormones from interacting with their receptors. TSP treatment markedly suppressed mammary phosphorylation levels of ERK1/2, as well as reduced the mRNA and protein overexpression of VEGF and bFGF in mammary of rats. In addition, TSP treatment substantially down-regulated the expression of Bcl-2 and Ki-67, as well as elevated the expression of Bax. These findings indicated that TSP could potentially be used for effective treatment of MGH.

Effects of puerarin on estrogen-regulated gene expression in gonadotropin-releasing hormone pulse generator of ovariectomized rats

Effects of puerarin on the hypothalamic gonadotropin-releasing hormone (GnRH) pulse generator function is investigated, for the first time, in ovariectomized rats at the level of mRNA expression of estrogen-responsive genes, e.g., estrogen receptor (ER), GnRH and its receptor (GnRHR). Rats were treated orally for 90 days either with a soy-free diet containing two different doses of puerarin (low dose of 600 mg/kg and high dose of 3000 mg/kg) or estradiol benzoate (E2B) at either low dose (4.3 mg/kg) or high dose (17.3 mg/kg). Levels of mRNA expression in the medial preoptic area/anterior hypothalamus (MPOA/AH), mediobasal hypothalamus/median eminence (MBH/ME) and adenohypophysis were measured by quantitative TaqMan® real-time RT-PCR. Plasma levels of luteinizing hormone (LH) and prolactin (PRL) were measured by radioimmunoassay. In the MPOA/AH, both puerarin and E2B decreased ERα mRNA levels without any significant changes in ERβ and GnRH mRNA levels. Both puerarin and E2B did not significantly alter the expression levels of ERα, ERβ and GnRHR in the MBH/ME. E2B exerted significant effects on the down-regulation of adenohypophyseal GnRHR mRNA transcripts and serum LH levels. Puerarin did not cause significant changes in pituitary GnRHR mRNA transcripts and serum LH and PRL levels. This is the first study to demonstrate that in ovariectomized rat models of ovarian hormone deprivation, puerarin acted as a weak estrogen-active compound in the hypothalamic GnRH pulse generator through the downregulation of MPOA/AH ERα mRNA expression.

Diminished androgen and estrogen receptors and aromatase levels in hypogonadal diabetic men: reversal with testosterone.

One-third of males with type 2 diabetes (T2DM) have hypogonadism, characterized by low total and free testosterone concentrations. We hypothesized that this condition is associated with a compensatory increase in the expression of androgen receptors (AR) and that testosterone replacement reverses these changes. We also measured estrogen receptor and aromatase expression. This is a randomized double-blind placebo-controlled trial. Thirty-two hypogonadal and 32 eugonadal men with T2DM were recruited. Hypogonadal men were randomized to receive intramuscular testosterone or saline every 2 weeks for 22 weeks. We measured AR, ERα and aromatase expression in peripheral blood mononuclear cells (MNC), adipose tissue and skeletal muscle in hypogonadal and eugonadal males with T2DM at baseline and after 22 weeks of treatment in those with hypogonadism. The mRNA expression of AR, ERα (ESR1) and aromatase in adipose tissue from hypogonadal men was significantly lower as compared to eugonadal men, and it increased significantly to levels comparable to those in eugonadal patients with T2DM following testosterone treatment. AR mRNA expression was also significantly lower in MNC from hypogonadal patients compared to eugonadal T2DM patients. Testosterone administration in hypogonadal patients also restored AR mRNA and nuclear extract protein levels from MNC to that in eugonadal patients. In the skeletal muscle, AR mRNA and protein expression are lower in men with hypogonadism. Testosterone treatment restored AR expression levels to that comparable to levels in eugonadal men. We conclude that, contrary to our hypothesis, the expression of AR, ERα and aromatase is significantly diminished in hypogonadal men as compared to eugonadal men with type 2 diabetes. Following testosterone replacement, there is a reversal of these deficits.

Luteolin inhibits ER-α expression through ILK inhibition is regulated by a pathway involving Twist and YB-1

Increasing evidence supports the anti-estrogenic and/or anti-proliferative properties of luteolin, which is a natural flavonoid existing in herbs, vegetables and fruits. However, the molecular mechanisms responsible for these effects are only partially explored. In the present study, estrogen receptor-alpha (ER-α) was identified as a potential molecular target for luteolin. Our data indicated that treatment with luteolin engendered suppression of ER-α protein and mRNA expression in ER-α-positive breast cancer cells. Y-box binding protein-1 (YB-1) is a known transcriptional regulator of ER-α expression. In this studies it was demonstrated that luteolin decreased YB-1 protein and mRNA expression. Integrin-linked kinase (ILK) is able to phosphorylate many downstream effectors that have the potential to regulate YB-1 transcription. We provided evidence for a mechanism by which luteolin suppressed YB-1 expression through the downregulation of ILK/Akt/STAT3-regulated Twist expression in ER-α-positive breast cancer cells. Our data also showed that luteolin inhibited cancer migration and invasion through the inhibition of epithelial-mesenchymal transition (EMT). Taken together, these data indicate that the antitumorigenic effects of luteolin in ER-α-positive breast cancer cells may arise from its ability to reduce ER-α expression.

Effect of side of the corpus luteum and pregnancy on estrogen and progesterone receptor expression and localization in the endometrium of mares

The effect of side of corpus luteum on uterine gene expression and protein localization of estrogen receptor α (ERα) and progesterone receptor (PR) in healthy cyclic and pregnant mares 13 days after ovulation (day 0) was investigated. Transcervical biopsies were performed to collect endometrium ipsilateral and contralateral regarding the side of corpus luteum on day 13 post-ovulation in cyclic (n = 6) and pregnant (n = 6) mares. Blood samples were collected daily from day 0 until the day of biopsy for 17β-estradiol (E2) and progesterone (P4) determinations. Receptor expression was determined by immunohistochemistry and transcript expression by real time RT-PCR. Serum E2 and P4 concentrations were not affected by reproductive status. The contralateral horn presented higher percentage of positive cells for ERα than the ipsilateral horn (P < .05), but side did not affect PR. ERα showed low staining and no main effect of pregnancy was found, but pregnant mares had lower protein expression of PR (19.8 vs. 40.4 ± 5.3%, P < .01). The contralateral horn tended to present higher expression of ERα mRNA (1.33 vs. 0.97 ± 0.17, P < .10) and PR mRNA (1.96 vs. 1.57 ± 0.52, P < .09). ERα mRNA relative expression was lower in the pregnant group (0.88 vs. 1.44 ± 0.19, P < .05). The interaction of reproductive status and side of corpus luteum tended to affect PR mRNA expression as pregnant mares had a lower PR mRNA content in the ipsilateral horn than cyclic mares. To our knowledge, this is the first study that describes the behavior of steroid receptors in the endometrium of mares regarding side of corpus luteum.

Expression of estrogen receptors in female rainbow trout (Oncorhynchus mykiss) during first ovarian development and under dense rearing condition

To study the expression of four estrogen receptor genes (erα1, erα2, erβ1, erβ2) of female rainbow trout (Oncorhynchus mykiss) during first ovarian development, trouts were sampled from different ovarian stages. Serum E2 (estradiol) was measured by ELISA and estrogen receptors mRNA expression were examined by qRT-PCR. Our results showed a close association between increased erα1 and vitellogenin mRNA expression during ovarian maturation and increased erα2 mRNA expression in mature ovarian stages. Correlation analysis revealed that a negative relationship between serum E2 and ovarian erβ1 (or hepatic erβ2), but ovarian erβ2 mRNA expression was relatively unchanged during first ovarian development. Trout were also reared in different densities as stocking density 1, 2 and 3 (SD1, 4.6–31.1 kg/m3; SD2, 6.6–40.6 kg/m3; SD3, 8.6–49.3 kg/m3) to elucidate effects of high density on estrogen receptor expression. Histology observation showed ovarian development of trout in higher densities were retard with a relatively early stage and fewer vitellogenin accumulation. Trout in high densities showed significantly decreased serum E2, erα mRNA expression and increasing trends of erβ mRNA expression. A noticeable increase of ovarian erβ2 mRNA expression was seen in trout when density is approaching to 50 kg/m3. In conclusion, we may hypothesize that increased erβ mRNA expression triggered by high density result in decreased erα mRNA expression and vitellogenesis. As a result, ovarian development in higher densities was retard.

Concomitant high expression of ER\u03b136, EGFR and HER2 is associated with aggressive behaviors of papillary thyroid carcinomas

ERα, ERβ, PR, ERα36, EGFR and HER2 mRNA and protein expression in papillary thyroid carcinoma (PTC) were examined by real time RT-PCR and immunohistochemical staining. The mRNA and protein expression of ERα and PR were gradually increased and those of ERβ were gradually decreased from normal thyroid tissues to nodular hyperplasias (P < 0.05) and to PTCs (P < 0.05). However, the mRNA and protein expression of ERα36, EGFR and HER2 were only significantly increased in PTCs when compared with those in normal thyroid tissues (P < 0.001) and nodular hyperplasias (P < 0.001). There was some correlation between ERα, ERβ and PR, and between ERα36, EGFR and HER2 protein expression in PTCs. As for ERα, ERβ and PR, there was a significant positive correlation between ERα and PR, and a significant negative correlation between ERα and ERβ and between PR and ERβ protein expression. As for ERα36, EGFR and HER2, there was a significant positive correlation between ERα36, EGFR and HER2 protein expression in PTCs. Concomitant high expression of ERα36, EGFR and HER2 was strongly associated with aggressive behaviors including extrathyroidal extension (ETE), lymph node metastasis (LNM) and high TNM stage in PTCs (P < 0.001).

Effects of electroacupuncture on uterine morphology and expression of oestrogen receptors in ovariectomised rats.

To observe the effects of electroacupuncture (EA) on uterine morphology and expression of oestrogen receptor (ER) α and β in ovariectomised (OVX) rats. Thirty female Sprague-Dawley rats with regular 4-day oestrus cycles were divided into a sham operation group (Control, n=10) and two OVX groups that remained untreated (OVX group, n=10) or received EA treatment (OVX+EA group, n=10). In the latter group, EA was applied at CV4, CV3, SP6 and bilateral Zigong (30 min per day) for 3 days. The effects of EA on uterine morphology were observed by H&E staining. Quantitative real-time reverse transcription-PCR (qRT-PCR) and Western blotting were used to measure ERα and ERβ mRNA and protein expression, respectively. Relative to the (untreated) OVX group, EA treatment significantly increased the uterine wet weight to body weight (UWW/BW) ratio (0.47±0.04 vs 0.31±0.03 g/kg, p=0.04), and myometrial thickness (109.39±10.71 vs 60.81±8.1 μm, p=0.016) of OVX rats. Similarly, the total number of endometrial glands per cross section and endometrial thickness in the OVX +EA group was significantly increased compared to the (untreated) OVX group. EA treatment also increased protein (but not mRNA) expression of both ERα and ERβ in the uteri of OVX rats. This study has demonstrated that EA treatment decreases uterine atrophy in OVX rats. This unique effect of EA on the uterus may be due to upregulation of serum levels of E2 and differential regulation of sex steroid receptors ERα and ERβ.

Seasonal changes of androgen receptor, estrogen receptors and aromatase expression in the hippocampus of the wild male ground squirrels (Citellus dauricus Brandt)

The wild ground squirrel is a typical seasonal breeder whose annual life cycle can be roughly divided into the breeding season, the post-breeding season and hibernation. Our previous study has reported the seasonal changes in the expressions of androgen receptor (AR), estrogen receptors α and β (ERα and ERβ), and aromatase cytochrome P450 (P450arom) in the hypothalamus of male wild ground squirrels. To further seek evidence of seasonal expression of steroid hormone receptors and steroid hormone synthases in other brain regions, we investigated the protein and mRNA expressions of AR, ERα, ERβ and P450arom in the hippocampus of the male wild ground squirrels during these different reproductive periods. Histological observation showed that the number of pyramidal cells in Cornu Ammonis 1 (CA1) increased in the breeding season. Both protein and mRNA of AR, ERα, ERβ and P450arom were present in CA1 and CA3 of all seasons. There was significant increment in the immune-signal intensity and mRNA level of AR and ERα during the pre-hibernation, whereas those of ERβ and P450arom were higher during the post-breeding season. In addition, the profile of plasma testosterone concentration showed the nadir in the post-breeding season, a marked elevation in the pre-hibernation, and the summit in the breeding season. These findings suggested that the hippocampus may be a direct target of androgen and estrogen; androgen may play important regulatory roles through its receptor and/or the aromatized estrogen in the hippocampus of the wild male ground squirrels.

Effect of growth hormone on steroid concentrations and mRNA expression of their receptor, and selected egg-specific protein genes in the chicken oviduct during pause in laying induced by fasting

This study was undertaken to examine the effect of growth hormone (GH) treatment during pause in laying on (1) the concentration of steroids in blood plasma and oviduct tissues, (2) the expression of mRNA of steroid receptors, and (3) the mRNA expression of selected egg-specific proteins in the chicken oviduct. A pause in egg laying was induced by food deprivation for 5 d, followed by feeding every other day, and then feeding daily from Day 10 onward. Birds were divided into three groups: control (n = 18) fed ad libitum, subjected to pause in laying (n = 18), and subjected to pause in laying and injected every day with 200 μg/kg BW of chicken GH (chGH; n = 18). The oviduct was isolated from hens of each group on Days 6 (when the oviduct was regressed), 13 (during oviduct recrudescence), and 17 or 20 (rejuvenated oviduct) of the experiment. Fasting caused a decrease in plasma concentrations of progesterone (P4), testosterone, and estradiol on Day 6 and a reduction in tissue concentrations of these steroids on Days 6 and 13. Fasting also caused an increased relative expression of estrogen receptor α and β (ERα, ERβ) and progesterone receptor (PR) in the magnum and shell gland on Day 6, increased ERα and PR in the magnum on Days 13 and 17 or 20, and increased androgen receptor (AR) mRNA in the magnum on Days 6 and 13 and in the shell gland on Day 13. A fasting-induced elevation in ovocalyxin-36 mRNA expression on Day 6 and a decrease in avidin mRNA on Days 6 and 13 and in ovocleidin-116 on Day 13 were also observed (P < 0.05 to P < 0.001). Administration of chGH abolished the fasting-induced decrease in the concentration of steroids in plasma and tissues. Furthermore, chGH enhanced the effect of fasting on mRNA expression of PR, ERα, and avidin in the magnum on Day 6, and ERα in the shell gland on Day 13. The gene expression of ovalbumin on Days 6 and 13, ovocalyxin-36 and ovocleidin-116 on Day 6 was decreased in chGH-treated chickens. In contrast, the expression of ovalbumin on Day 17 or 20 was increased (P < 0.05 to P < 0.001). The results obtained indicate that, by alterations in the concentration of steroid hormones and their receptor expression in the chicken oviduct, GH determines the rate of regression and rejuvenation of this organ during molting. Moreover, changes in the expression of selected egg proteins indicate that GH might be the regulator of the secretory activity of the hen oviduct.

ERα and GnRH co-localize in the hypothalamic neurons of the South American plains vizcacha, Lagostomus maximus (Rodentia, Caviomorpha).

Gonadotropin-releasing hormone (GnRH) is the key regulator of the hypothalamic-pituitary-gonadal axis. Estradiol (E2) affects GnRH synthesis and delivery. Hypothalamic estrogen receptors (ER) modulate GnRH expression acting as transcription factors. The South American plains vizcacha, Lagostomus maximus, is able to ovulate up to 800 oocytes per reproductive cycle, and shows continuous folliculogenesis with pre-ovulatory follicle formation and an ovulatory event at mid-gestation. The aim of this work was to analyze the hypothalamic expression of ER in the vizcacha at different gestational time-points, and its relationship with GnRH expression, serum luteinizing hormone (LH) and E2. The hormonal pattern of mid-gestating vizcachas was comparable to ovulating-females with significant increases in GnRH, LH and E2. Hypothalamic protein and mRNA expression of ERα varied during pregnancy with a significant increase at mid-gestation whereas ERβ mRNA expression did not show significant variations. Hypothalamic immunolocalization of ERα was observed in neurons of the diagonal band of Brocca, medial preoptic area (mPOA), periventricular, suprachiasmatic, supraoptic (SON), ventromedial, and arcuate nuclei, and medial eminence, with a similar distribution throughout gestation. In addition, all GnRH neurons of the mPOA and SON showed ERα expression with no differences across the reproductive status. The correlation between GnRH and ERα at mid-gestation, and their co-localization in the hypothalamic neurons of the vizcacha, provides novel information compared with other mammals suggesting a direct action of estrogen as part of a differential reproductive strategy to assure GnRH synthesis during pregnancy.

Age- and menopause-related differences in subcutaneous adipose tissue estrogen receptor mRNA expression

ObjectivesChanges in estrogen receptor (ER) expression likely underlie differential metabolic effects of estrogen in pre- and postmenopausal women. The aim of the current study was to determine whether ER gene expression in abdominal and femoral subcutaneous adipose tissue (SAT) was associated with age, menopause, or regional adiposity. MethodsWe studied pre- and post-menopausal (n=23 and 22, respectively; age 35–65y) normal weight (mean±SD; BMI 23.7±2.5kg/m2) women with similar total fat mass. Abdominal and femoral SAT ERα (ESR1) and ERβ (ESR2) mRNA expression was determined by qPCR. ResultsTotal fat mass did not differ between pre- and postmenopausal women (22.7±5.3vs. 21.7±5.3kg). Compared to premenopausal women, ESR1 and the ratio of ESR1 to ESR2 were lower (p≤0.05) in postmenopausal abdominal and femoral SAT. ESR1 and ESR1:ESR2 were inversely associated with age in abdominal SAT (r=−0.380 and r=−0.463, respectively; p<0.05) and femoral SAT (r=−0.353 and r=−0.472, respectively; p<0.05). ESR2 was not related to age or menopause. The inverse association between ESR1 and age persisted after adjusting for trunk fat mass, estradiol, or leptin. ConclusionAmong healthy pre- and postmenopausal women, increased age was associated with a decreased balance of ERα to ERβ in abdominal and femoral subcutaneous adipose tissue.

Expression of ER and PR in the endometrium of patients with intrauterine adhesions

Objective: To estimate the expression of ER and PR in the endometrium of both intrauterine adhesions (IUA) and non-IUA specimens. Methods: The endometrium specimens from patients undergoing hysteroscopy for confirmed moderate IUA (n=20: 10 in proliferative phase, and 10 in secretory phase) were enrolled as the IUA group in Beijing Obstetrics and Gynecology Hospital from October 2014 to August 2015. The specimens scheduled for hysteroscopy due to infertility were recruited into the control group (n=26: 13 in proliferative phase, and 13 in secretory phase). Immunohistochemistry and quantificational real-time PCR (qRT-PCR) were used to detect the expression of ER-α, ER-β and PR in endometrium with different menstrual period in both groups. Results: (1) Location: in both groups, the expression of ER-α, ER-β and PR appeared in the endometrial glandular epithelial cells and the stromal cells of the endometrium. The positive brown granules of ER-α, ER-β and PR appeared mainly in cell nucleus. (2) ER-α and ER-β in the endometrium: the protein expression of ER-α and ER-β in IUA group (proliferative phase: 0.657±0.028, 0.493±0.023; secretory phase: 0.537±0.020, 0.365±0.031) were significantly higher than those of control group (proliferative phase: 0.586±0.025, 0.437±0.022; secretory phase: 0.459±0.025, 0.323±0.017; all P<0.01). And the ER-α and ER-β mRNA expressions in IUA group were 2.524±0.296, 1.947±0.339, higher than those of control group in the proliferative phase (all P<0.01), and in the secretory phase (1.977±0.333, 1.345±0.292) were also higher than those in the control group (all P<0.01). (3) PR in the endometrium: the protein expression of PR was not significantly different between IUA group (proliferative phase: 0.248±0.025, secretory phase: 0.194±0.024) and control group (proliferative phase: 0.234±0.019, secretory phase: 0.186±0.020; P=0.162, 0.359). Meanwhile, there were no statistical differences in the mRNA expression of PR in both groups with different menstrual period (proliferative phase: 1.144±0.384 versus 0.981±0.306, secretory phase: 0.763±0.237 versus 0.631±0.203; P=0.270, 0.166). (4) ER and PR expression in menstrual cycles: the expression of ER-α, ER-β and PR in the IUA group changed with the menstrual cycles, and their expression in the proliferative phase were higher than those in the secretory phase (all P<0.05). Conclusions: The expression of ER-α and ER-β in the endometrium of IUA patients changes with menstrual cycle, and are higher compared with those in normal endometrium. No difference is found in the PR expression between the two groups.

Developmental changes in hypothalamic SF-1, POMC, and ERα mRNA expression and their sensitivity to fasting in male and female rats.

Estrogen plays pivotal roles in body weight regulation through its effects on central estrogen receptor-α (ERα) expression. ERα is found on neurons that express the hypothalamic anorexigenic factors steroidogenic factor-1 (SF-1) and pro-opiomelanocortin (POMC) mediate these effects of estrogen. As the gonadal hormonal milieu is drastically altered during the developmental period, the expression levels of SF-1 and POMC might also change during this period. In this study, we showed that hypothalamic SF-1 and ERα mRNA expression did not change during the neonatal to pre-pubertal period (from postnatal day 10 to 30), and there were no significant differences in the hypothalamic mRNA expression levels of these molecules between males and females at any examined age. On the other hand, hypothalamic POMC mRNA expression and the serum estradiol (E2) level increased during development in both males and females. Significant positive correlations were detected between the serum E2 level and hypothalamic POMC mRNA expression in both males and females. Hypothalamic ERα and POMC mRNA expression were decreased by fasting in male rats at all examined ages, whereas fasting had no effect on hypothalamic ERα or POMC mRNA expression in the female rats. These results indicate that the regulatory system involving E2 and hypothalamic POMC expression might already be established in the neonatal period and that the roles of POMC and ERα in body weight regulation during development might differ slightly between males and females.

A potential mechanism associated with lead-induced testicular toxicity in rats.

This study was conducted to investigate the mechanism of lead (Pb)-induced testicular toxicity. We examined the impact of Pb toxicity on 17β-oestradiol (E2), oestrogen receptors (ERs) and aromatase P450 which are key factors in spermatogenesis. Treatment of rats with Pb acetate (PbAc, 50mg/L in drinking water) significantly reduced sperm count, motility, viability and increased sperm abnormalities along with degenerative changes in seminiferous tubules and Leydig cells. Additionally, administration of PbAc resulted in a significant reduction in serum testosterone, serum and testicular E2 as well as increased level of testicular testosterone. Pb also induced testicular oxidative stress as evidenced by a significant decrease in the activities of superoxide dismutase, glutathione peroxidase and catalase antioxidant enzymes, and increased malondialdehyde level in the testis. At the molecular level, Pb treatment downregulated the mRNA expression of P450 arom (Cyp19) and ERα. In conclusion, Pb induces testicular oxidative damage and disrupts spermatogenesis, at least in part, via downregulation of Cyp19 and ERα expression, which further decrease E2 level. These data, therefore, provide insight into the mechanism of lead-induced testicular toxicity.

Correlation of BCL-2 and ERα mRNA Expression with the Clinical Chemotherapeutic Response in Breast Cancer

Colonic angiodysplasia is now recognized as a frequent cause of gastrointestinal bleeding in aged patient. The lesions are not evident on barium studies. Even colonoscopy and selective colonic angiography may give false negative results. Therefore, repeated studies for recurrent episodes of bleeding are often necessary. There are a lot of methods of treatment which includes hormonal therapy, electrocoagulation, laser or local sclerosing agent injection for few and small lesions (less than 1cm) and surgical treatment for large and extensive lesions.We report a young man, aged 35, presented with chronic anemia due to lower gastrointestinal bleeding, was found to have several small actively oozing angiodysplasias over the transverse colon near the hepatic flexure during colonoscopic examination. Another 3 small lesions without bleeding were also found over rectum. Endoscopic injection therapy for hemostasis with pure alcohol was performed successfully in all lesions. Follow-up colonoscopy one month later revealed no more bleeding and some scars formation were detected. The hemoglobin (Hgb) was stable at about 12g/dl and no more fecal occult blood has been detected in the 6 months follow-up peroid. Endoscopic local injection therapy is an effective, safe and convenient therapy for small angiodysplasias in the colon.