雌激素受体亚型在人乳腺癌的表达及意义

目的　研究人乳腺癌组织以及癌旁正常乳腺组织中雌激素受体 (ER)亚型ERα和ERβ的表达 ,及其在乳腺癌发生发展中的作用。方法　应用逆转录聚合酶链反应方法检测 30例患者乳腺癌组织以及相应癌旁正常组织中ERα和ERβ的mRNA的表达情况。结果　ERα在人乳腺癌组织的表达明显高于癌旁正常组织 (t=7 399,P <0 0 1) ,ERβ在人乳腺癌组织的表达明显低于癌旁正常组织 (t=- 3 2 36 ,P <0 0 1) ,ERα/ERβ比值在人乳腺癌组织明显高于癌旁正常组织 (t =6 385 ,P <0 0 1) ;没有淋巴结转移的乳腺癌患者ERα/ERβ比值明显高于有淋巴结转移的乳腺癌患者 (t =2 6 0 2 ,P <0 0 5 ) ;在分期较晚的肿瘤ERα/ERβ比值明显低于分期较早的肿瘤 (t =3 75 4 ,P <0 0 5 )。结论　ERα在乳腺癌发生发展过程中与ERβ发挥不同的作用 ,并有可能成为乳腺癌基因治疗的新靶点。     

DNA聚合酶θ对乳腺癌细胞株MCF-7放射敏感性的影响

目的 观察DNA聚合酶θ(POLQ)对人类乳腺癌细胞株MCF-7放射敏感性的影响.方法 实时荧光定量聚合酶链反应(qRT-PCR)和Western blot法检测siRNA转染前后POLQ表达率的变化;利用噻唑蓝(MTT)比色法、平板克隆形成实验、流式细胞术对不同实验组进行分析.结果 POLQ-siRNA组细胞POLQ mRNA和POLQ蛋白表达量分别是空白对照组的(12.16±0.56)％和(32.01±0.65)％ (P＜0.01).转染后72 h,POLQ-siRNA组、空白对照组细胞吸光度(A)值分别为(0.76±0.02)和(0.81 ±0.02),POLQ-siRNA 组细胞增殖能力明显低于空白对照组(P＜0.01);平板克隆形成实验分析结果显示POLQ-siRNA组放射增敏比(SER)为1.24(＞1),表明转染POLQ-siRNA后MCF-7细胞放射敏感性显著增强.结论 抑制POLQ表达可以显著提高MCF-7细胞对放射的敏感性.     

腺病毒介导的靶向survivin的siRNA上调乳腺癌MCF-7细胞雌激素受体表达

目的 探讨腺病毒介导的靶向survivin的小干扰RNA(siRNA)对乳腺癌MCF-7细胞雌激素受体(ER)表达的影响.方法 构建靶向survivin的siRNA腺病毒载体并转染乳腺癌MCF-7细胞,采用Western blot法检测MCF-7细胞转染腺病毒后survivin、ER表达的变化.结果 实验组、阴性组、空白组survivin的表达强度分别为0.09 ±0.04、0.86±0.08、0.82 ±0.17;ER的表达强度分别为1.57 ±0.09、1.16±0.10、1.23±0.01.统计学分析表明,本研究中所构建的靶向survivin的siRNA腺病毒载体可以高效地抑制survivin的表达,抑制survivin的表达可以显著上调乳腺癌MCF-7细胞ER的表达(P＜0.05).结论 在乳腺癌MCF-7细胞中,survivin与ER信号通路之间可能存在着某种相互调节机制,提示靶向survivin的siRNA在ER阳性乳腺癌的内分泌治疗中可能具有潜在的重要意义.     

雌激素受体β与乳腺癌

目的 探讨雌激素受体β（ERβ）在乳腺癌方面的研究进展，方法 采用文献回顾方法。对ERβ的生物学功能，变异，及其乳腺癌发生，发展，预后评估及内分泌治疗反应方面的可能作用加以综述。结果 ERβ是类固醇激素受体超家族的新成员，可能在乳腺癌发生，发展，预后评估及内分泌治疗反应方面具有重要作用。结论 ERβ是一种新的乳腺癌预后评估指标。     

雌激素受体阴性孕激素受体阳性乳腺癌研究进展

乳腺癌病理学亚型可根据其免疫组化结果如雌激素受体（estrogen receptor,ER）、孕激素受体（progesterone receptor,PR）、人表皮生长因子受体2（human epidermal growth factor receptor2,HER2）及Ki67表达进一步细分为不同的分子亚型,包括管腔...     

乳腺癌雌激素受体研究进展

三苯氧胺是目前乳腺癌内分泌治疗最常用的一线药物，但部分却对三苯氧治疗不起反应。本文对雌激素受体的结构，功能，研究进展等方面作一简要介绍。     

雌激素受体在人乳腺癌MCF-7细胞阿霉素敏感株和耐药株中的变化及意义

目的研究MCF 7人乳腺癌细胞阿霉素敏感株 (MCF 7)和耐药株 (MCF 7/ADR)细胞中雌激素受体 (estrogenreceptor,ER)表达的变化与其对细胞生物学特性的影响。方法采用Westernblot法检测MCF 7细胞和MCF 7/ADR细胞中雌激素受体表达 ,RT PCR检测 2株细胞ERmRNA的表达水平 ,MTT法检测细胞增殖及细胞对雌激素 (estrogen ,E2 )和屈洛昔芬 (droloxifene ,Dro)的敏感性 ,流式细胞仪检测细胞周期变化。结果经Westernblot检测 ,MCF 7细胞ER为阳性 ,而MCF 7/ADR细胞ER为阴性 ,采用RT PCR可检测到MCF 7细胞中ER的mRNA ,而在MCF 7/ADR细胞则检测不到。经MTT分析 ,与MCF 7相比 ,MCF 7/ADR细胞生长速度减慢 ,细胞多分布于G0 /G1期。E2 在 1× 10 -12mol/L的浓度时开始促进MCF 7细胞的生长 ,到达 1× 10 -9mol/L时促生长作用进入平台期 ;而任何浓度的E2 对MCF 7/ADR细胞均无促生长作用。Dro的浓度达到 10 μmol/L的时候 ,开始出现对MCF 7细胞生长的浓度依赖性抑制 ,当Dro浓度达到 2 0 μmol/L时对MCF 7/ADR细胞的生长出现明显抑制 ,且高于对MCF 7细胞的抑制。结论MCF 7/ADR细胞雌激素受体表达缺失 ,缺失发生在mRNA水平以上 ,细胞生长速度减慢 ,同时细胞失去对雌激素的依赖性。     

乳腺癌雌激素受体研究进展

三苯氧胺是目前乳腺癌内分泌治疗最常用的一线药物，但部分却对三苯氧治疗不起反应。本文对雌激素受体的结构、功能、研究进展等方面作一简要介绍。     

乳腺癌他莫昔芬耐药与雌激素受体β亚型表达的关系

目的 探讨雌激素受体亚型β(ERβ)及其剪切变异体ERβcx表达与乳腺癌他莫昔芬治疗耐药的关系。方法 将MCF-7细胞、MCF-7/TAM-R细胞(他莫昔芬耐药株)以及5-氮杂胞苷(5-AZA-CdR)去甲基化作用后的MCF-75-AZA细胞和MCF-7/TAM-R5-AZA细胞采用Western blot检测ERα、ERβ和ERβcx蛋白的表达;将MCF-7细胞作为对照,用他莫昔芬干预上述细胞,噻唑蓝(MTT)比色法观察细胞增殖,流式细胞仪检测细胞周期变化。结果 ERα的表达在各组间无明显差异;ERβ的表达差异有统计学意义(P＜0.01),在MCF-7/TAM-R组细胞中最低,在MCF-75-AZA组中最高;MCF-7和MCF-7/TAM-R组的ERβcx表达差异无统计学意义(P＞0.05),但明显低于MCF-75-AZA和MCF-7/TAM-R5-AZA组的表达(P＜0.01)。MTT检测结果显示,与对照组比较,他莫昔芬干预后MCF-7/TAM-R 细胞增殖速度差异无统计学意义(P＞0.05);而MCF-7、MCF-75-AZA和MCF-7/TAM-R5-A细胞生长均受到抑制,其中MCF-75-AZA细胞受抑程度强于MCF-7细胞(P＜0.01),而MCF-7/TAM-R5-AZA细胞受抑制的程度低于MCF-7细胞(P＜0.01)。流式细胞仪测定这3组细胞的S期比率都明显下降。结论 ERβ蛋白表达和他莫昔芬治疗内分泌治疗敏感相关,ERβcx蛋白表达与耐药无关,但可能与ERβ共同影响乳腺癌对他莫昔芬治疗的敏感性。5-AZA-CdR去甲基化可以诱导乳腺癌细胞ERβ及其剪切异构体ERβcx表达增强,改善他莫西芬治疗效果。     

三氧化二砷诱导乳腺癌细胞系MCF-7凋亡的实验研究

目的观察不同浓度的三氧化二砷(As2O3)作用不同时间后对乳腺癌细胞系MCF-7的影响及其作用机理。方法应用不同浓度的As2O3作用于乳腺癌细胞后,观察As2O3对乳腺癌细胞生长状态的影响;用四甲基偶氮唑蓝(MTT)比色法观察其对MCF-7细胞增殖的影响;应用琼脂糖凝胶电泳和脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(TUNEL)检测As2O3对MCF-7细胞凋亡的诱导作用。结果不同浓度的As2O3作用于乳腺癌细胞有明显的时间和剂量依赖性,As2O3作用后细胞生长明显受抑制,并出现明显的凋亡特征性改变(细胞膜完整、染色质固缩、核碎裂、凋亡小体形成);经1、2、4及8μmol/L4个剂量的As2O3处理48h后,琼脂糖凝胶电泳检测到细胞发生凋亡时DNA降解形成梯形分子条带;4μmol/LAs2O3作用24、48及72h后TUNEL法可检测到细胞凋亡水平显著性升高。结论As2O3可明显抑制乳腺癌细胞的生长,其机理主要是诱导乳腺癌细胞凋亡。     

VEGF-C基因转染对人乳腺癌MCF-7细胞VEGF-C表达的影响

目的探讨将真核表达载体pcDNA3．1-VEGF—C重组质粒转染入人乳腺癌MCF-7细胞后VEGF-C表达的变化。方法通过脂质体介导方法，将构建好含有VEGF—C的重组质粒转染入人乳腺癌MCF-7细胞中，新霉素（G418）筛选得到稳定转染细胞系，RT—PCR和Western blot方法检测稳定转染后细胞中VEGF—C mRNA和蛋白的表达。结果成功转染并获得稳定高表达VEGF—C的乳腺癌MCF-7细胞系，其转染组VEGF—C mRNA相对吸光度值（12．382&#177;2．183）较空载组（6．039&#177;1．950）显著上调（P〈0．01）。Western blot检测转染组VEGF-C蛋白相对灰度值（0．971&#177;0．186）较空载组（0．594&#177;0．196）明显上调（P〈0．05）。结论脂质体介导重组质粒pcDNA3．1-VEGF-C转染入人乳腺癌MCF7细胞中可显著增加VEGF—C表达水平。     

Estrogen Receptor Alpha in Human Breast Cancer: Occurrence and Significance

Estrogens have long been recognized as being important for stimulating the growth of a large proportion of breast cancers. Now it is recognized that estrogen action is mediated by two receptors, and the presence of estrogen receptor (ER)³ correlates with better prognosis and the likelihood of response to hormonal therapy. Over half of all breast cancers overexpress ER and around 70% of these respond to anti-estrogen (for example tamoxifen) therapy. In addition, the presence of elevated levels of ER in benign breast epithelium appears to indicate an increased risk of breast cancer, suggesting a role for ER in breast cancer initiation, as well as progression. However, a proportion of ER-positive tumors does not respond to endocrine therapy and the majority of those that do respond eventually become resistant. Most resistant tumors remain ER-positive and frequently respond to alternative endocrine treatment, indicative of a continued role for ER in breast cancer cell proliferation. The problem of resistance has resulted in the search for and the development of diverse hormonal therapies designed to inhibit ER action, while research on the mechanisms which underlie resistance has shed light on the cellular mechanisms, other than ligand binding, which control ER function.     

柠檬苦素类化合物对人乳腺癌细胞(MCF-7)的生长抑制及细胞周期动力学的影响

为了探讨柠檬苦素类似物对乳腺癌细胞MCF-7生长抑制和对细胞周期的影响，应用MTT法和流式细胞仪检测癌细胞株生长和细胞周期.结果显示柠檬苦素类似物能明显抑制MCF-7的生长，抑制率高于70%，并随作用时间和剂量增加其效果增强；柠檬苦素可以影响细胞周期G     

The Loss of Estrogen and Progesterone Receptor Gene Expression in Human Breast Cancer

Hormone responsiveness is a critical determinantof breast cancer progression and management, and theresponse to endocrine therapy is highly correlated withthe estrogen receptor (ER)³ and progesterone receptor (PR) status of tumor cells. Thus, keyareas of study in breast cancer are those mechanismsthat regulate ER and PR expression in normal andmalignant breast tissues. One-third of all breastcancers lack ER and PR; these conditions are associatedwith less differentiated tumors and poorer clinicaloutcome. In addition, approximately one-half ofER-positive tumors lack PR protein and patients withthis phenotype are less likely to respond tohormonal therapies than those whose tumors express bothreceptors. Since PR is induced by ER; its presence is amarker of a functional ER. In this review, we will discuss possible mechanisms for loss of ER andPR gene expression, especially structural changes withineach gene including deletions, polymorphisms ormethylation. Improved understanding of the pathways that lead to loss of ER and/or PR proteinsshould allow the development of better predictiveindicators as well as novel therapeutic approaches totarget these hormone-independent cancers.     

华蟾素诱导人乳腺癌细胞株MCF-7细胞凋亡与Bax/Bcl-2的关系

目的:观察华蟾素在诱导人乳腺癌MCF-7细胞株凋亡的过程中Bax和Bcl-2蛋白表达的变化及意义。方法:在人乳腺癌细胞株MCF-7中分别加入不同终浓度的华蟾素,24h、48h及72h后检测指标。应用MTT比色法检测细胞增殖活力,倒置显微镜观察细胞形态学变化,TUNEL法检测细胞凋亡指数,Western blot法检测Bax/Bcl-2蛋白表达,共聚焦显微镜观察Bax蛋白分布。结果:华蟾素能显著抑制MCF-7细胞增殖;倒置显微镜下发现凋亡的MCF-7细胞固缩、核染色质凝聚;western blot发现华蟾素能上调MCF-7细胞Bax蛋白表达,但对Bcl-2蛋白表达无影响,正常MCF-7细胞内的Bax均匀分布于细胞质中,华蟾素处理后,Bax明显地聚集于线粒体,呈现线粒体的转位。结论:华蟾素诱导MCF-7细胞凋亡,并在该过程中导致的Bax蛋白表达上调并且向线粒体转位。     

In Vivo Modeling of Human Breast Cancer Using Cell Line and Patient-Derived Xenografts

Historically, human breast cancer has been modeled largely in vitro using long-established cell lines primarily in two-dimensional culture, but also in three-dimensional cultures of varying cellular and molecular complexities. A subset of cell line models has also been used in vivo as cell line-derived xenografts (CDX). While outstanding for conducting detailed molecular analysis of regulatory mechanisms that may function in vivo, results of drug response studies using long-established cell lines have largely failed to translate clinically. In an attempt to address this shortcoming, many laboratories have succeeded in developing clinically annotated patient-derived xenograft (PDX) models of human cancers, including breast, in a variety of host systems. While immunocompromised mice are the predominant host, the immunocompromised rat and pig, zebrafish, as well as the chicken egg chorioallantoic membrane (CAM) have also emerged as potential host platforms to help address perceived shortcomings of immunocompromised mice. With any modeling platform, the two main issues to be resolved are criteria for “credentialing” the models as valid models to represent human cancer, and utility with respect to the ability to generate clinically relevant translational research data. Such data are beginning to emerge, particularly with the activities of PDX consortia such as the NCI PDXNet Program, EuroPDX, and the International Breast Cancer Consortium, as well as a host of pharmaceutical companies and contract research organizations (CRO). This review focuses primarily on these important aspects of PDX-related research, with a focus on breast cancer.

     

Breast Cancer Subtypes as Defined by the Estrogen Receptor (ER), Progesterone Receptor (PR), and the Human Epidermal Growth Factor Receptor 2 (HER2) among Women with Invasive Breast Cancer in California, 1999–2004

Abstract: Breast cancer research examining either molecular profiles or biomarker subtypes has focused on the estrogen receptor negative/progesterone receptor negative/human epidermal growth factor receptor 2 negative (ER−/PR−/HER2−) and ER−/PR−/HER2+ subtypes. Less is known about the epidemiology or clinical outcome of the other subtypes. This study examines the eight combinations of ER/PR/HER2 in patients with invasive breast cancer. The 5‐year relative survival and the distribution among demographic, socioeconomic, and tumor characteristics of each of the subtypes are examined. Using the California Cancer Registry, 61,309 women with primary invasive breast cancer were classified according to ER/PR/HER2 status. Five‐year relative survival was computed for the eight subtypes. Bivariate analyses were used to assess the distribution of cases across all subtypes. Multivariate logistic regression was used to compute the adjusted odds of having one of the five subtypes with the best and worst survival. Survival varied from 96% (ER+/PR+/HER2−) to 76% (ER−/PR−/HER2+ and ER−/PR−/HER2−). The four subtypes with the poorest survival were all ER negative. Women who were younger than age 50, non‐Hispanic black or Hispanic, of the lowest SES groups, and had stage IV tumors that were undifferentiated were overrepresented in ER−/PR−/HER2+ and triple negative (ER−/PR−/HER2−) subtypes. Asian Pacific Islanders had increased odds (OR = 1.41; 95% confidence interval [CI] = 1.26–1.57) of having the ER−/PR−/HER2+ subtype. Stage III tumors (OR = 1.25; 95% CI = 1.08–1.44) and stage IV tumors (OR = 1.58; 95% CI = 1.27–1.98) had higher odds than stage I tumors of being ER−/PR−/HER2+. Stage IV tumors (OR = 0.54; 95% CI = 0.44–0.67) strongly decreased the odds of the ER−/PR−/HER2− subtype. Poorly differentiated and undifferentiated tumors were over 20 times as likely as well‐differentiated tumors of being ER−/PR−/HER2− or ER−/PR−/HER2+. There are considerable differences in survival, demographics, and tumor characteristics among the eight subtypes. We recommend reporting breast cancer as an ER/PR/HER2 subtype and precisely documenting demographic and tumor characteristics.