BPH-1通过分泌PGE2上调前列腺间质细胞ERRα的表达

摘要 雌激素受体相关受体α（estrogen receptor-related receptor α,ERRα）是一类可以直接或间接参与雌激素应答反应的孤儿核受体,它与雌激素受体（estrogen receptor）在结构上有很强的同源性．雌激素效应在良性前列腺增生（benign prostatic hyperplasis,BPH）的发生和发展中起着重要的作用．通常,孤儿核受体的转录活性多受一些非经典激素如维生素A衍生物、前列腺素类、固醇的调控．本文研究前列腺上皮细胞分泌的活性因子对间质细胞ERRα表达调控的分子机制．收集前列腺增生上皮细胞系BPH-1和前列腺癌上皮细胞系DU-145的条件培养液（condition medium,CM）培养的间质细胞,采用实时定量RT-PCR和Western 印迹法检测前列腺间质细胞（prostate stromal cells,PrSCs）中ERRα的表达,筛选CM中影响ERRα表达的活性因子．研究结果显示,BPH-1的CM可以上调ERRα的表达,而DU-145的CM对ERRα的表达没有影响;BPH-1中合成前列腺素E2 (prostaglandin E2, PGE2)的限速酶——环氧合酶2（cyclooxygenase-2, COX-2）的mRNA表达水平和PGE2的分泌水平明显高于DU-145中COX-2表达水平和PGE2分泌水平;用经添加COX-2抑制剂NS-398的培养液处理BPH-1,其CM中PGE2的浓度明显下降,并失去了对ERRα表达的上调作用;添加PGE2可上调间质细胞中ERRα的表达．结果表明,BPH-1通过分泌PGE2促进间质细胞ERRa的表达,提示：在良性前列腺增生的发生和发展中,上皮细胞的旁分泌作用可促进间质细胞由ERRα介导的雌激素效应．     

雌激素受体相关受体亚型在鼻咽癌中的表达及分析

目的:探讨雌激素受体相关受体(estrogen receptor related receptors,ERR)α、β、γ亚型表达与鼻咽癌的关系.方法:随机抽取健康体格合者40人为时照组,鼻咽癌患者40例为试验组,采用RT-PCR方法检测ERRs家族各亚型的mRNA表达,采用Westston Blot印迹方法检测ERR各亚型在鼻咽癌组织中的表达.结果:RT-PCR及Western Blot蛋白印记检测显示ERR-α在对照组低表达、在实验组高表达,两组患者外周血及鼻咽组织中ERR-B表达差异无统计学意义,ERR-γ在对照组高表达、在实验组低表达.结论:ERR-α和ERR-γ在鼻咽癌患者体内表达异常,其可能与鼻咽癌的发生发展相关.     

新型雌激素受体ER-α36与乳腺癌

雌激素受体(estrogen receptor,ER)是诊断和治疗乳腺癌的分子标志和靶点.雌激素受体包括ER-α和ER-β,其中ER-α有ER-α66、ER-α46和ER-α36三种亚型.ER-α36作为新型雌激素受体,参与膜起始的雌激素信号或非基因组雌激素信号转导,在肿瘤细胞的增殖、分化、侵袭和转移等过程中发挥作用.胃癌、子宫内膜腺癌、前列腺癌、尤其是乳腺癌的发生发展与ER-α36密切相关.本文介绍了ER-α36的结构域特点,ER-α36介导的信号通路及ER-α36在乳腺癌治疗中的作用研究进展.     

肌钙蛋白I2的原核表达及与ERRα1的体外相互作用

孤儿受体雌激素受体相关受体α1(ERRα1)与乳腺癌有密切的关系.酵母双杂交筛选发现乳腺组织中肌钙蛋白I2(TNNI2)与其有明显的相互作用.为进一步研究TNNI2与ERRα1的相互作用,融合表达了GST-TNNI2原核蛋白,并体外翻译了35S标记的ERRα1蛋白,将二者共同温育进行GST捕获实验.放射自显影结果显示,TNNI2能够捕获35S标记的ERRα1,证明TNNI2与ERRα1存在体外直接的相互结合,提示了肌钙蛋白I2在ERRα1参与的生理过程中可能有重要的作用.     

雌激素受体ERα的功能调控及相关疾病的研究进展

雌激素受体(estrogen receptorα,ERα)是依赖配体活化转录因子的核受体家族成员之一,参与靶细胞的增殖和分化。ERα活化的经典途径是与雌激素结合后直接作用于靶基因上游的雌激素受体反应元件(ERE),从而诱导靶基因转录。雌激素受体的功能受许多因子调节,包括与之结合的配体、DNA上的顺式元件、募集的辅助调节因子及细胞环境等。在雌激素受体相关疾病中,除乳腺癌和子宫内膜癌外,近年研究表明心血管疾病、骨质疏松症、阿尔茨海默氏病等疾病也与雌激素受体密切相关。雌激素的生物效应与多种疾病的发生、转归和预后密切相关。本文将综述几类辅助调节因子对雌激素受体介导的基因转录的调控,雌激素受体相关疾病,及环境有害物质对ERα功能的影响。     

雌激素受体α36

雌激素受体(estrogen receptor,ER)属核受体超家族成员之一,主要包括ER-α66、ER-α36、ER-α46和ER-β,它们通过与相应配体——雌激素的结合,在人体中发挥各种不同的功能。ER-α36是最近新发现的一种雌激素受体,它是ER-α66的独特变异体,主要分布于细胞膜和胞浆中,能抑制ER-α66和ER-β的反式激活功能,并参与雌激素的非基因组活性信号通路,介导了临床乳腺癌抗雌激素药物治疗抵抗。     

雌激素受体阳性乳腺癌内分泌治疗耐药机制及靶向治疗策略

乳腺癌是中国女性最常见的恶性肿瘤之一。内分泌治疗是雌激素受体(estrogen receptor, ER)阳性乳腺癌有效的治疗手段之一,也是最常用的方法,主要包括选择性雌激素受体调节剂(selective estrogen receptor modulators, SERMs)、芳香化酶抑制剂(aromatase inhibitors, AIs)、选择性雌激素受体下调剂(selective estrogen receptor down regulators, SERDs)。尽管内分泌治疗增加了患者生存率,但耐药问题也极大地影响了乳腺癌患者的预后。越来越多的耐药机制被报道,如ESR1突变、受体酪氨酸激酶改变、细胞信号通路、表观遗传调控、代谢重编程、肿瘤干细胞、肿瘤微环境改变等。该文对近几年报道的乳腺癌内分泌治疗耐药机制及靶向治疗策略进行详细的综述。     

ERR促进乳腺癌发生的作用机制

乳腺癌是发病率逐年增加、女性最常见的恶性肿瘤之一.ERR(雌激素受体相关受体)α,β和γ(NR3B1-3)是核受体超家族成员,也是配体依赖的转录因子.ERRs可通过线粒体、血管生成、脂肪生成等方式参与乳腺癌肿瘤细胞的代谢.ERRs的功能不受配体影响,但受特异性共调控子调节.这些调控子包括类固醇受体的共激活体(SRC)、PPARγ共激活体PGC-1α/β和共抑制体受体相互作用蛋白140(RIP140).对ERR的研究可为治疗乳腺癌提供新的途径.     

肌钙蛋白I2辅助活化多种核受体的反式作用

为深入研究乳腺癌中孤儿受体ERRα1参与基因表达调控的详细机理,特别是核受体辅激活蛋白在其中的作用,以ERRα1的LBD为诱饵,用酵母双杂交系统筛选人乳腺组织cDNA文库得到了与其有明显相互作用的快速骨骼肌型肌钙蛋白I(TNNI2).应用酵母双杂交技术研究表明,TNNI2与多种核受体存在相互作用,且这种作用依赖于功能性的核受体AF2结构域.在哺乳细胞瞬时共转染实验中,TNNI2显示了对多种核受体反式激活功能的辅助活化作用.研究证明,TNNI2与许多辅激活蛋白类似,以配体依赖(对类固醇激素受体而言)或非依赖(对孤儿受体而言)的方式与核受体功能性AF2结构域相互作用,并增强多种核受体介导的反式作用.     

乳腺癌相关的细胞内信号通路

乳腺癌是女性中常见的恶性肿瘤之一.乳腺癌的发生、发展、转移及耐药性的产生与细胞内的信号通路密切相关,其中雌激素受体(estrogen receptor,ER)信号通路、胰岛素样生长因子受体（insulin-like growth factor receptor,IGFR)信号通路和表皮生长因子受体（epidermal growth factor receptor,EGFR）信号通路尤为重要．深入了解ER、IGFR和EGFR三条信号通路的作用机制及它们之间的交叉对话对于寻找新的更有效的肿瘤治疗靶点至关重要．本文综述了近年来有关ER、IGFR和EGFR三条信号通路研究进展及这三条通路与乳腺癌关系．     

雄激素和雌激素受体药物筛选方法的研究进展

雄激素和雌激素受体通过与相应激素特异性结合促进细胞分化和组织生长,发挥重要的生理功能,其功能失调可诱发多种疾病。雄激素和雌激素受体的选择性调节剂是治疗相关疾病的重要药物。基于基因组学、分子生物学、细胞生物学和生物信息学等最新研究成果而发展形成的实验技术或方法被用于新型雄激素和雌激素受体调节剂的筛选,显著加快了新药开发的进程。     

Analysis of 3D models of octopus estrogen receptor with estradiol: evidence for steric clashes that prevent estrogen binding

Relatives of the vertebrate estrogen receptor (ER) are found in Aplysia californica, Octopus vulgaris, Thais clavigera, and Marisa cornuarietis. Unlike vertebrate ERs, invertebrate ERs are constitutively active and do not bind estradiol. To investigate the molecular basis of the absence of estrogen binding, we constructed a 3D model of the putative steroid-binding domain on octopus ER. Our 3D model indicates that binding of estradiol to octopus ER is prevented by steric clashes between estradiol and amino acids in the steroid-binding pocket. In this respect, octopus ER resembles vertebrate estrogen-related receptors (ERR), which have a ligand-binding pocket that cannot accommodate estradiol. Like ERR, octopus ER also may have the activation function 2 domain (AF2) in a configuration that can bind to coactivators in the absence of estrogens, which would explain constitutive activity of octopus ER.     

Substituted phenyl as a steroid A-ring mimetic: Providing agonist activity to a class of arylsulfonamide nonsteroidal glucocorticoid ligands

A class of arylsulfonamide glucocorticoid receptor agonists that contains a substituted phenyl group as a steroid A-ring mimetic is reported. The structural design and SAR that provide the functional switching of a GR antagonist to an agonist is described. A combination of specific hydrogen bonding and lipophilic elements on the A-ring moiety is required to achieve potent GR agonist activity. This study culminated in the identification of compound 23 as a potent GR agonist with selectivity over the PR and MR nuclear hormone receptors.     

The ecdysteroid receptor

Summary

The steroid molting hormone of insects and other arthropods regulates gene activity in target tissues through its association with a specific, high affinity receptor protein. In this review we summarize recent advances in several areas of ecdysteroid receptor research, including efforts to characterize and purify the receptor protein, cytochemical studies of its tissue distribution and subcellular localization during development, and current molecular genetic studies ecdysteroid action.     

降钙素基因相关肽家族受体及其受体活性修饰蛋白

降钙素基因相关肽家族是一类多功能的激素家族 ,参与人体的多种生物学功能 ,与多种疾病有关。降钙素基因相关肽受体包括降钙素受体 (CTR)和降钙素受体样受体 (CRLR) ,CTR可以独自与降钙素结合 ,而CRLR必须与一组称作受体活性修饰蛋白 (RAMPs)的蛋白质共同作用才能发挥生物学功能。综述CTR的研究概况及CRLR与RAMPs相互作用的机制和表达调控 ,以期为人们设计新型药物提供参考。     

Structural Investigations of Full-Length Insulin Receptor Dynamics and Signalling

Insulin regulates glucose homeostasis via binding and activation of the insulin receptor dimer at two distinct pairs of binding sites 1 and 2. Here, we present cryo-EM studies of full-length human insulin receptor (hIR) in an active state obtained at non-saturating, physiologically relevant insulin conditions. Insulin binds asymmetrically to the receptor under these conditions, occupying up to three of the four possible binding sites. Deletion analysis of the receptor together with site specific peptides and insulin analogs used in binding studies show that both sites 1 and 2 are required for high insulin affinity. We identify a homotypic interaction of the fibronectin type III domain (FnIII-3) of IR resulting in tight interaction of membrane proximal domains of the active, asymmetric receptor dimer. Our results show how insulin binding at two distinct types of sites disrupts the autoinhibited apo-IR dimer and stabilizes the active dimer. We propose an insulin binding and activation mechanism, which is sequential, exhibits negative cooperativity, and is based on asymmetry at physiological insulin concentrations with one to three insulin molecules activating IR.     

Ago-Allosteric Modulation and Other Types of Allostery in Dimeric 7TM Receptors

Conventionally, an allosteric modulator is neutral in respect of efficacy and binds to a receptor site distant from the orthosteric site of the endogenous agonist. However, recently compounds being ago-allosteric modulators have been described i.e., compounds acting both as agonists on their own and as enhancers for the endogenous agonists in both increasing agonist potency and providing additive efficacy—superagonism. The additive efficacy can also be observed with agonists, which are neutral or even negative modulators of the potency of the endogenous ligand. Based on the prevailing dimeric concept for 7TM receptors, it is proposed that the ago-allosteric modulators bind in the orthosteric binding site, but–importantly–in the “other” or allosteric protomer of the dimer. Hereby, they can act both as additive co-agonists, and through intermolecular cooperative effects between the protomers, they may influence the potency of the endogenous agonist. It is of interest that at least some endogenous agonists can only occupy one protomer of a dimeric 7TM receptor complex at a time and thereby they leave the orthosteric binding site in the allosteric protomer free, potentially for binding of exogenous, allosteric modulators. If the allosteric modulator is an agonist, it is an ago-allosteric modulator; if it is neutral, it is a classical enhancer. Molecular mapping in hetero-dimeric class-C receptors, where the endogenous agonist clearly binds only in one protomer, supports the notion that allosteric modulators can act through binding in the “other” protomer. It is suggested that for the in vivo, clinical setting a positive ago-allosteric modulator should be the preferred agonist drug.     

Proteinase-activated receptors, nucleotide P2Y receptors, and μ-opioid receptor-1B are under the control of the type I transmembrane proteins p23 and p24A in post-Golgi trafficking

We recently characterized the proteinase-activated receptor (PAR)-2, a G protein-coupled receptor (GPCR), as the first cargo protein recognized by p24A. Here, we demonstrate that p24A binds to several other GPCRs, including PAR-1, the nucleotide receptors P2Y(1), P2Y(2), P2Y(4), and P2Y(11), as well as the μ-opioid receptor 1B. The acidic amino acid residues Glu and Asp at the second extracellular loop of GPCRs are essential for interaction with p24A. p23, another member of the p24 family, also interacts with GPCRs, similar to p24A. However, p23 shows a delayed dissociation from PAR-2 after activation of PAR-2, compared to the dissociation between PAR-2 and p24A. p24A and p23 arrest both P2Y(4) receptor and μ-opioid receptor 1B at the intracellular compartments, as observed for PAR-2. A comparable result was obtained when we studied primary rat astrocytes in culture. Over-expression of the N-terminal p24A fragment impairs PAR-2 resensitization in astrocytes that extends our findings to a native system. In summary, we demonstrate that p24A and p23 are specific cargo receptors of GPCRs and differentially control GPCR trafficking in the biosynthetic pathway, and thereby, p24A and p23 regulate GPCR signaling in astrocytes.     

新型抗抑郁药物分子靶标研究进展

抑郁症是一种严重的精神障碍疾病,其发病机制复杂。近年来随着对抑郁症发病机制的深入研究,发现了一些基于非单胺递质的 新型抗抑郁药物分子靶标。综述N -甲基-D-天冬氨酸（NMDA）受体、促肾上腺皮质激素释放因子（CRF）受体、阿片受体、γ-氨基丁 酸B(GABAB) 受体、乙酰胆碱受体等抗抑郁药物作用的新靶标及其相应分子机制研究进展,为开发高效、安全的抗抑郁症新药提供参考。