Aggressive Prostate Cancer Is Prevented in ERαKO Mice and Stimulated in ERβKO TRAMP Mice

Previous evidence suggests soy genistein may be protective against prostate cancer, but whether this protection involves an estrogen receptor (ER)-dependent mechanism is unknown. To test the hypothesis that phytoestrogens may act through ERα or ERβ to play a protective role against prostate cancer, we bred transgenic mice lacking functional ERα or ERβ with transgenic adenocarcinoma of mouse prostate (TRAMP) mice. Dietary genistein reduced the incidence of cancer in ER wild-type (WT)/transgenic adenocarcinoma of mouse prostate mice but not in ERα knockout (KO) or ERβKO mice. Cancer incidence was 70% in ERWT mice fed the control diet compared with 47% in ERWT mice fed low-dose genistein (300 mg/kg) and 32% on the high-dose genistein (750 mg/kg). Surprisingly, genistein only affected the well differentiated carcinoma (WDC) incidence but had no effect on poorly differentiated carcinoma (PDC). No dietary effects have been observed in either of the ERKO animals. We observed a very strong genotypic influence on PDC incidence, a protective effect in ERαKO (only 5% developed PDC), compared with 19% in the ERWT, and an increase in the incidence of PDC in ERβKO mice to 41%. Interestingly, immunohistochemical analysis showed ERα expression changing from nonnuclear in WDC to nuclear in PDC, with little change in ERβ location or expression. In conclusion, genistein is able to inhibit WDC in the presence of both ERs, but the effect of estrogen signaling on PDC is dominant over any dietary treatment, suggesting that improved differential targeting of ERα vs. ERβ would result in prevention of advanced prostate cancer.     

雌激素受体亚型ERα、ERβ在乳腺癌中的表达

目的研究雌激素受体亚型ERα、ERβ在不同乳腺组织中的表达及其在浸润性乳腺癌中与肿瘤病理参数和生物学标志物之间的关系。方法选取70例乳腺癌(其中8例原位癌、44例浸润性导管癌、18例浸润性小叶癌)、15例乳腺增生、15例正常乳腺组织石蜡标本,用免疫组织化学法检测ERα、ERβ、C-erbB-2的表达情况。结果 (1)乳腺癌组织中的ERα阳性表达率明显高于正常乳腺及乳腺增生组织;晚期乳腺癌患者ERα阳性表达率明显低于早期患者;C-erbB-2阳性患者的ERα阳性表达率明显减低。(2)乳腺癌组织中的ERβ阳性表达率明显低于正常乳腺及乳腺增生组织;在浸润性乳腺癌中淋巴结转移阳性者、C-erbB-2表达阳性者的ERβ阳性表达率明显高于对照组患者。结论 ERα在乳腺癌的发生发展中可能起到促进作用,ERα的阳性表达与一些预后良好因素相关。ERβ可能对维持乳腺组织的正常生理功能起着重要作用,其与某些预后不良因素可能相关。     

胰腺癌中ERα变异体的表达

目的检测胰腺癌中雌激素受体α(ERα)不同变异体ERC4、ERD3、ERD5 mRNA的表达.方法采用免疫组化和RT-PCR检测技术,对50例胰腺癌和20例乳腺癌患者ERα的表达以及变异体ERC4、ERD3、ERD5 mRNA的表达进行检测.结果胰腺癌ERα蛋白的阳性率为32%.ERD3 mRNA在ER阳性胰腺癌表达低于阴性病例;ERC4 mRNA在ER阴性胰腺癌表达明显低于阳性病例(P < 0.05);ERD5 mRNA在ER阳性胰腺癌表达低于阴性病例.结论 ERα变异体的检出说明它们在胰腺癌发生过程中起一定的作用.在胰腺癌的发生过程中,ERα变异体mRNA的表达失控.     

胰腺癌中ERα变异体的表达

目的检测胰腺癌中雌激素受体α(ERα)不同变异体ERC4、ERD3、ERD5mRNA的表达。方法采用免疫组化和RT-PCR检测技术,对50例胰腺癌和20例乳腺癌患者ERα的表达以及变异体ERC4、ERD3、ERD5mRNA的表达进行检测。结果胰腺癌ERα蛋白的阳性率为32％。ERD3mRNA在ER阳性胰腺癌表达低于阴性病例;ERC4mRNA在ER阴性胰腺癌表达明显低于阳性病例(P<0.05);ERD5mRNA在ER阳性胰腺癌表达低于阴性病例。结论ERα变异体的检出说明它们在胰腺癌发生过程中起一定的作用。在胰腺癌的发生过程中,ERα变异体mRNA的表达失控。     

Occurrence and cellular distribution of estrogen receptors ERα and ERβ in the testis and epididymal region of roosters

Estrogen signaling is required for the maintenance of male reproductive function and is mediated by the estrogen receptors ERα and ERβ. These receptors are widely distributed in mammalian reproductive tissues, but information is limited in non-mammalian species including birds. The aim of this study was to investigate the occurrence and cellular distribution of ERα and ERβ in the testis and epididymal region of roosters. The results showed for the first time that ERβ was the predominant receptor detected in the testis, being expressed in the somatic and some germ cells. Within the epididymal region, ERβ was strongly expressed in all segments, whereas the most intense reaction for ERα was found in the distal efferent ductules. The differential expression of ERα and ERβ within the rooster testis and epididymal region suggests that these organs may be a target for different actions of estrogen.     

High ERα36 Expression Level and Membrane Location Predict Poor Prognosis in Renal Cell Carcinoma

Estrogen receptor alpha 36 (ERα36), a truncated variant of ERα, is located in cytoplasm and membrane that is different from other nuclear receptors of ERα family. ERα36 is involved in progression and treatment resistance of a variety of carcinomas. However, the clinical and prognostic significance of ERα36 in renal tumors have not been fully elucidated.Here, renal tumor tissues from 125 patients were collected and immunohistochemical stained with ERα36 antibody. ERα36 expression level and location in these cases were analyzed for their correlations with clinical characteristics. The differential diagnosis value was also assessed for benign and malignant renal tumors, as well as its prognostic value.The results showed that membrane ERα36 expression was rarely detected in benign tumors but predominantly observed in malignant renal tumors. Kaplan–Meier analysis indicated that significant correlations of high ERα36 level and ERα36 membrane expression were correlated with both poor disease-free survival and overall survival. Univariate and multivariate analysis confirmed that both ERα36 high expression and membrane location can serve as unfavorable prognostic indicators for renal cell carcinoma.It is thus concluded that membrane ERα36 expression is valuable for differential diagnosis of malignant renal tumors from benign ones. Both ERα36 high expression and membrane location indicate poor prognosis in renal cell carcinoma.     

Ratio of Circulating Estrogen Receptors Beta and Alpha (ERβ/ERα) Indicates Endoscopic Activity in Patients with Crohn’s Disease

Background

Data supporting a role of female hormones and/or their receptors in inflammatory bowel disease (IBD) are increasing, but most of them are derived from animal models. Estrogen receptors alpha (ERα) and beta (ERβ) participate in immune and inflammatory response, among a variety of biological processes. Their effects are antagonistic, and the net action of estrogens may depend on their relative proportions.

Aim

To determine the possible association between the balance of circulating ERβ and ERα (ERβ/ERα) and IBD risk and activity.

Methods

Serum samples from 145 patients with IBD (79 Crohn’s disease [CD] and 66 ulcerative colitis [UC]) and 39 controls were retrospectively studied. Circulating ERα and ERβ were measured by ELISA. Disease activities were assessed by clinical and endoscopic indices specific for CD and UC.

Results

Low values of ERβ/ERα ratio were directly associated with clinical (p = 0.019) and endoscopic (p = 0.002) disease activity. Further analyses by type of IBD confirmed a strong association between low ERβ/ERα ratio and CD clinical (p = 0.011) and endoscopic activity (p = 0.002). The receiver operating curve (ROC) analysis showed that an ERβ/ERα ratio under 0.85 was a good marker of CD endoscopic activity (area under the curve [AUC]: 0.84; p = 0.002; sensitivity: 70%; specificity: 91%). ERβ/ERα ratio was not useful to predict UC activity.

Conclusions

An ERβ/ERα ratio under 0.85 indicated CD endoscopic activity. The determination of serum ERβ/ERα might be a useful noninvasive screening tool for CD endoscopic activity.

     

Seasonal variation in estrogen receptor ERα, but not ERβ, androgen receptor and aromatase, in the efferent ductules and epididymis of the big fruit-eating bat Artibeus lituratus

The efferent ductules (ED) are a major target for estrogens, which act via the estrogen receptors ERα (ESR1) and ERβ (ESR2). ERα has been found in the ED of all species studied so far. However, in the epididymis (EP), the expression of ERα is controversial, as is data about the occurrence of aromatase in the epithelium lining the excurrent ducts. Therefore, to further investigate this estrogen-responsive system, we used a seasonal breeder, the Neotropical bat, Artibeus lituratus, in which testicular expression of androgen (AR) and estrogen (ER) receptors vary with reproductive phase. The localization of aromatase, ERα, ERβ and AR in the ED and EP of A. lituratus was investigated. The results showed that aromatase, AR and ERβ were distributed throughout the excurrent ducts and did not vary during the annual reproductive cycle. Conversely, ERα was detected primarily in the ED epithelium, had marked seasonal variation and was increased during regression, especially in the EP epithelium. The results suggest that ERα may be involved in preparing the male genital tract for recrudescence. Together, the data obtained under natural conditions emphasize that specific segments of the excurrent ducts downstream of the testis are the primary targets for estrogen action via ERα, which is similar to previous findings in animals lacking functional ERα.     

Interactions between IGF-I, estrogen receptor-α (ERα), and ERβ in regulating growth/apoptosis of MCF-7 human breast cancer cells

Understanding of the interactions between estradiol (E?) and IGF-I is still incomplete. Cell lines derived from the MCF-7 breast cancer cells were generated with suppressed expression of the IGF-I receptor (IGF-IR), termed IGF-IR.low cells, by stable transfection using small interfering RNA (siRNA) expression vector. Vector for control cells carried sequence generating noninterfering RNA. Concomitant with reduction in the IGF-IR levels, the IGF-IR.low cells also showed a reduction in estrogen receptor α (ERα) and progesterone receptor expressions, and an elevation in the expression of ERβ. The number of the IGF-IR.low cells was reduced in response to IGF-I and human GH plus epidermal growth factor, but E? did not cause an increase in the number of the IGF-IR.low cells compared to controls. The proliferation rate of IGF-IR.low cells was only reduced in response to E? compared to controls, whereas their basal and hormone-stimulated apoptosis rate was increased. Phosphorylation of p38 mitogen-activated protein kinase (p38 MAPK) was increased in the IGF-IR.low cells after treatment with E?, without affecting control cells. Furthermore, phosphorylation of the tumor suppressor protein p53 was elevated in the IGF-IR.low cells compared to the controls. In conclusion, suppressing IGF-IR expression decreased the level of ERα but increased the level of ERβ. Overall growth rate of the IGF-IR.low cells was reduced mostly through an increase in apoptosis without affecting proliferation substantially. We hypothesize that a decreased ERα:ERβ ratio triggered a rapid phosphorylation of p38 MAPK, which in turn phosphorylated the p53 tumor suppressor and accelerated apoptosis rate.     

ERαKO小鼠缺血模型中雌激素的作用机制研究

目的通过对ER-α受体缺乏小鼠大脑中动脉梗死模型(MCAO)的研究,进一步了解雌激素的作用机制.方法用生理盐水、25 μg/mL及50μg/mL β-雌二醇分别处理雄性野生型小鼠(WT)和α基因敲除型小鼠(ERαKO),制成MCAO模型后进行行为学评分,缺血2 h后再灌注22 h取脑测定梗死面积.结果WT中雌激素处理后的小鼠梗死面积明显减少(P＜0.05),ERαKO小鼠中雌激素同样有效(P＜0.05).结论雌激素对缺血性脑组织损伤具有保护作用,去除雌激素α受体后并不影响雌激素的脑保护作用.     

Identification of estradiol/ERα-regulated genes in the mouse pituitary

Estrogen acts to prime the pituitary prior to the GnRH-induced LH surge by undiscovered mechanisms. This study aimed to identify the key components that mediate estrogen action in priming the pituitary. RNA extracted from the pituitaries of metestrous (low estrogen) and proestrus (high estrogen) stage mice, as well as from ovariectomized wild-type and estrogen receptor α (ERα) knockout mice treated with 17β-estradiol (E(2)) or vehicle, was used for gene expression microarray. Microarray data were then aggregated, built into a functional electronic database, and used for further characterization of E(2)/ERα-regulated genes. These data were used to compile a list of genes representing diverse biological pathways that are regulated by E(2) via an ERα-mediated pathway in the pituitary. This approach substantiates ERα regulation of membrane potential regulators and intracellular vesicle transporters, among others, but not the basic components of secretory machinery. Subsequent characterization of six selected genes (Cacna1a, Cacna1g, Cited1, Abep1, Opn3, and Kcne2) confirmed not only ERα dependency for their pituitary expression but also the significance of their expression in regulating GnRH-induced LH secretion. In conclusion, findings from this study suggest that estrogen primes the pituitary via ERα by equipping pituitary cells with critical cellular components that potentiate LH release on subsequent GnRH stimulations.     

ERα基因多态性与Alzheimer病智能衰退的相关分析

目的 探讨中国汉族人群中雌激素受体(ER)α基因酶切多态性与散发性Alzheimer氏病(SAD)的发病及其智能衰退的相关性.方法 采用聚合酶链反应限制片段长度多态性方法,检测63例AD患者及70例健康对照老年人的ERαPvuⅡ、XbaⅠ酶切基因多态性;采用MMSE、ADL量表观察38例AD患者6个月的智能衰退状况与ERα基因多态性的关系.结果 AD组ERα等位基因x的频率、Pp、xx、Ppxx型较对照组明显增加,而PP、XX、PPXX型明显减少(P＜0.05);XbaI酶切基因多态性xx、Ppxx与AD发病明显正相关(OR分别为2.34,95%CI 1.47～4.09和 1.79,95%CI 1.26～3.12)(P＜0.05).Pp与AD发病无明显相关(OR 1.58,95%CI 1.15～2.12).ERа基因xx、Ppxx型与AD的智能衰退无明显相关性.结论 雌激素α受体xx或Ppxx基因型是SAD发病的危险因素,但与AD的智能衰退无明显相关性;ERαXbaⅠ基因酶切多态性xx型可能与中国汉族SAD的发病具有相关性.     

ER stress signalling through eIF2α and CHOP, but not IRE1α, attenuates adipogenesis in mice

Aims/hypothesis

Although obesity is associated with endoplasmic reticulum (ER) stress and activation of the unfolded protein response (UPR) in adipose tissue, it is not known how UPR signalling affects adipogenesis. To test whether signalling through protein kinase RNA-like ER kinase/eukaryotic initiation factor 2 alpha (PERK/eIF2α) or inositol-requiring enzyme 1 alpha/X-box binding protein 1 (IRE1α/XBP1) is required for adipogenesis, we studied the role of UPR signalling in adipocyte differentiation in vitro and in vivo in mice.

Methods

The role of UPR signalling in adipogenesis was investigated using 3T3-L1 cells and primary mouse embryonic fibroblasts (MEFs) by activation or inhibition of PERK-mediated phosphorylation of the eIF2α- and IRE1α-mediated splicing of Xbp1 mRNA. Body weight change, fat mass composition and adipocyte number and size were measured in wild-type and genetically engineered mice fed a control or high-fat diet (HFD).

Results

ER stress repressed adipocyte differentiation in 3T3-L1 cells. Impaired eIF2α phosphorylation enhanced adipocyte differentiation in MEFs, as well as in mice. In contrast, increased eIF2α phosphorylation reduced adipocyte differentiation in 3T3-L1 cells. Forced production of CCAAT/enhancer binding protein (C/EBP) homologous protein (CHOP), a downstream target of eIF2α phosphorylation, inhibited adipogenesis in 3T3-L1 cells. Mice with deletion of Chop (also known as Ddit3) (Chop ^?/?) gained more fat mass than wild-type mice on HFD. In addition, Chop deletion in genetically obese Lepr ^db/db mice increased body fat mass without altering adipocyte size. In contrast to the eIF2α–CHOP pathway, activation or deletion of Ire1a (also known as Ern1) did not alter adipocyte differentiation in 3T3-L1 cells.

Conclusions/interpretation

These results demonstrate that eIF2α–CHOP suppresses adipogenesis and limits expansion of fat mass in vivo in mice, rendering this pathway a potential therapeutic target.     

α- andβ-mannosidoses

Summary Clinical, pathological and biochemical findings in the mannosidoses are described. Family studies showed granulocyte-rich white cell fractions to be the tissue of choice for carrier detection in-mannosidosis. Metabolic labelling studies using [³H] mannose demonstrated accumulation of Man1-4GlcNAc in cultured skin fibroblasts from a patient with this condition. Alternative methods of egress from lysosomes were suggested for this compound by its secretion into culture medium and apparent reduction of storage with time in cultures.-mannosidase deficient goats are not thought to be a true animal model of the human condition, as although they showed a similar enzyme deficiency, the clinical presentation is much more severe and the major storage material (Man1-4GlcNAc1-4GlcNAc) is different.     

Distribution of estrogen receptors (ERα and ERβ) and androgen receptor in the testis of big fruit-eating bat Artibeus lituratus is cell- and stage-specific and increases during gonadal regression

The testis is a classical target for androgens, especially testosterone, acting via androgen receptor (AR). Alternatively, androgens can be aromatized to produce estrogens which act via specific receptors ERα and ERβ. Although estrogen action is essential for maintenance of male fertility, studies regarding the expression of ERα and ERβ in testis are restricted to a few species of rodent and domestic animals, but rarely in wild species. To our knowledge, there are no studies in Chiroptera species. Chiroptera represent one of the largest and most diversified orders of mammals, which possess several interesting reproductive features, including higher affinity of SHBG for estrogens than androgens. Therefore, we thought that bats would constitute a good model for investigation of the role of estrogens in the male. In this study, the distribution of ERα, ERβ and AR were evaluated in the testis of the big fruit-eating bat Artibeus lituratus and their levels were compared during reproductive and regressive periods. The results showed that ERα and AR were restricted to the somatic cells of the testis, whereas ERβ was widely distributed in both somatic and spermatogenic cells in a cellular and stage-specific fashion. We demonstrated for the first time by immunohistochemistry, and confirmed by Western blotting, that ERβ and AR increased during regression. The localization of ERα, ERβ and AR in a seasonal, cell and stage-specific fashion in the testis of A. lituratus suggests that these receptors may play important roles in testis function during reproductive and non-reproductive periods.     

ERα-29位多态性与HBV感染相关原发性肝癌的关系

目的探讨雌激素受体(ER)α-29位多态性与乙型肝炎病毒(HBV)感染相关原发性肝癌(PHC)的关系。方法HBV感染相关PHC患者150例作为研究组,另选取同期健康体检志愿者100例作为对照组。检测并对比两组ERα-29基因型和等位基因,分析ERα-29等位基因中发生HBV感染相关PHC的危险性。结果研究组TT型、CC型、TC型比例分别为48例(32.00%)、37例(24.67%)、65例(43.33%),对照组TT型、CC型、TC型比例分别为12例(12.00%)、41例(41.00%)、47例(47.00%),两组基因型整体比较差异有统计学意义(P<0.05);研究组T、C等位基因占比分别为53.67%、46.33%,对照组分别为35.50%、64.50%,两组等位基因占比比较差异有统计学意义(P<0.05)。T等位基因出现HBV感染相关PHC的风险为C等位基因的2.542倍(OR=2.542,95%CI:1.423~3.958)。结论 HBV感染相关PHC患者和正常人血清内ERα-29多态性表达存在明显的差异,HBV感染相关PHC患者中以T等位基因高表达为主要特点,ERα-29位T等位基因可增加HBV感染相关PHC发病的风险。     

儿童氟斑牙与雌激素受体基因(ERα)Xbal多态性关系分析

<正>氟斑牙又称氟牙症或者斑釉牙,是因儿童在牙齿发育时期从食物、饮水、甚至空气中接触过多的氟所导致儿童牙釉质发育不全,是一种慢性氟中毒的早期症状[1]。随着地方性氟中毒发病机制研究的发展,对高氟水平对骨损伤的分子机制已经成为慢性氟中毒研究的重点[2]。在同样地高氟环境下,不同儿童因存在遗传易感性的差异,其耐受性也存在一定差异。近年研究发现,雌激素对骨骼发育有十分明显的影响,雌激素受体(ER)的基因多态性对雌激素活性的影响显     

ERα、EGFR 及 CyclinD1在上皮性卵巢癌中的表达及意义

目的：探讨雌激素受体α（ ERα）、表皮生长因子受体（ EGFR）及细胞周期蛋白D1（ CyclinD1）在上皮性卵巢癌中的表达及临床意义。方法采用免疫组化SP法观察ERα、EGFR及CyclinD1在上皮性卵巢癌组织58份（浆液性卵巢癌组织38份、非浆液性卵巢癌组织20份）、正常卵巢组织18份、卵巢良性肿瘤组织20份中的表达情况。结果 ERα、EGFR及CyclinD1在上皮性卵巢癌组织中的阳性表达率分别为74．13％、68．97％、51．72％,显著高于正常卵巢组织（33．33％、16．67％、0）及卵巢良性肿瘤组织（30．0％、25．0％、10．0％）（P均＜0．05）;但ERα、EGFR及CyclinD1在卵巢良性肿瘤、正常卵巢组织中的阳性表达率比较P均＞0．05。 ERα、EGFR及CyclinD1在上皮性卵巢癌中的表达与临床分期及细胞分化程度有关（P均＜0．05）。结论 ERα、EGFR及CyclinD1在上皮性卵巢癌中过表达,并在一定程度上反映其恶性程度及预后状况。     

HER-2、PTEN蛋白及ERα在子宫内膜癌中的表达及其临床意义

目的检测子宫内膜癌组织中HER-2、PTEN蛋白及雌激素受体亚型(ERα)表达并探讨其临床意义。方法采用免疫组织化学法检测60例子宫内膜癌、32例正常子宫内膜组织中HER-2、PTEN蛋白及ERα的表达。结果在正常的子宫内膜HER-2蛋白为低表达,子宫内膜癌中HER-2的表达率明显增高(P0.05),而且HER-2的表达与肿瘤组织学分级、病理学分期以及肿瘤浸润子宫肌层深度显著相关(P0.05),其表达与组织学类型、有无淋巴结转移无关(P0.05)。子宫内膜癌组织中PTEN缺失率高于正常子宫内膜组织(P0.05)。PTEN表达在G1级肿瘤高于G2、G3级(P0.05),PTEN蛋白缺失率与肿瘤组织类型有关(P0.05),与肌层浸润、淋巴结转移及病理分期无明显关系(P0.05)。HER-2与PTEN的表达呈呈明显负相关。ERα在子宫内膜癌表达低于正常子宫内膜,且随着病理分期表达显著降低(P0.05)。结论 PTEN表达缺失与临床病理参数无关,蛋白表达缺失常发生细胞分化较差的子宫内膜癌。PTEN蛋白表达缺失与HER-2表达水平有关。