雌激素受体α和β在不同雌激素干预大鼠骨代谢中的表达

应用雌性大鼠的骨质疏松模型,通过骨密度(BMD)检测、RT-PCR和Westernblot等技术观察去卵巢(Ovariectomy,OVX)、结合性雌激素(ConjugatedEquineEstrogens,CEE)和戊酸雌二醇(EstradiolValerate,EV)对大鼠松质骨骨量以及松质骨中雌激素受体(ER)α和βmRNA和蛋白表达的影响,探讨两受体亚型在介导雌激素参与松质骨代谢的不同作用机制以及不同来源雌激素对ERα和ERβ表达的差异性调节。40只7-8周龄的雌性Sprague-Dawley大鼠,在观察动情周期后随机分成四组:对照组(Control,n=10)、去卵巢组(Ovariectomy,OVX,n=10)、去卵巢后结合性雌激素治疗组(CEE,n=10)和去卵巢后戊酸雌二醇治疗组(EV,n=10)。对照组大鼠行假手术,其余三组行去卵巢手术。术后48天(12个动情周期),对照组和OVX组用生理盐水喂养12天(3个动情周期),CEE组和EV组分别用药物的生理盐水溶液喂养12天。结果显示:在对照组中,大鼠松质骨ERα的蛋白表达水平显著性高于ERβ蛋白表达水平,而ERα的mRNA表达水平显著性低于ERβmRNA水平。与对照组相比,OVX组大鼠松质骨中ERα的蛋白表达水平显著性降低,ERαmRNA表达水平显著性增加,而ERβ蛋白和mRNA的表达水平均显著性增加。与OVX组相比,CEE组大鼠松质骨中ERβ蛋白和mRNA的表达水平均显著性下降,而EV组大鼠松质骨中ERα蛋白表达显著性上升,ERαmRNA表达显著性下降,ERβ蛋白表达显著性下降。此外,OVX大鼠松质骨的骨密度下降均可通过应用CEE和EV得到显著性改善。上述结果提示:⑴ERα可能是大鼠松质骨中优势表达的受体亚型,在介导雌激素参与松质骨代谢中起着主导作用。⑵不同来源雌激素可能侧重不同的ER亚型途径产生骨保护效应。     

不同疗法对去卵巢大鼠子宫ERα和ERRα蛋白表达的影响

目的:本研究旨在对比观察全身垂直振动、跑台运动、金雀异黄酮和氯化锂等不同干预疗法对去卵巢骨质疏松大鼠子宫雌激素受体α(ERα)和雌激素受体相关受体α(ERRα)蛋白表达的影响。方法:将80只3月龄雌性SD大鼠按体重分层后随机分为假手术组和去卵巢组。去卵巢10周时,将去卵巢组大鼠按体重分层后又随机分为去卵巢组、振动组、跑台组、金雀异黄酮组、氯化锂组和雌激素组,并开始进行不同的干预处理。干预处理8周时,腹主动脉取血处死各组大鼠,用放射免疫方法检测血清E_2水平,用Western blot检测子宫ERα和ERRα蛋白表达水平的变化。结果:大鼠去卵巢后,血清E_2水平显著下降,经雌激素处理后,血清E_2水平显著回升,但经其他几种方法处理后均无显著变化。Western blot结果显示,大鼠去卵巢后,子宫ERα蛋白表达水平显著增加,经雌激素、跑台运动、全身垂直振动和氯化锂处理后,子宫ERα蛋白表达水平显著下降,而经金雀异黄酮处理后,子宫ERα蛋白表达水平无显著变化。大鼠去卵巢后,子宫ERRα蛋白表达水平显著下降,经雌激素和金雀异黄酮干预后,子宫ERR-α表达水平显著增加,而经跑台运动、全身振动和氯化锂干预后,子宫ERR-α表达水平却显著下降。结论:跑台运动和全身垂直振动能抑制去卵巢骨质疏松大鼠子宫ERα和ERRα蛋白的表达,氯化锂能抑制ERα蛋白的表达,但促进ERRα蛋白的表达,金雀异黄酮能促进ERRα蛋白的表达,但对ERα蛋白的表达没有影响。     

性激素对大鼠诱发肝癌中胎盘型谷胱甘肽S-转移酶(GST-P)表达的影响

本实验利用Solt-Farber顺序诱发大鼠肝癌,观察肝组织中GST活性及GST-P含量在化学诱癌中的变化,并观察性激素对大鼠化学诱发肝癌的早期病变中GST-P表达的作用。结果显示无论是GST活性或GST-P的含量,在诱癌至第三周开始升高,第五周升至最高。利用此模式,选择诱癌至第五周,免疫组化法检测各种处理后肝组织中GST-P的表达。发现睾丸假切除的雄性大鼠经化学诱癌后,肝中有高的GST-P表达,睾丸假切除的雄性大鼠诱癌合并雌二醇处理,明显降低肝组织GST-P阳性灶的面积和数量;合并睾丸酮处理,虽减少GST-P阳性灶的面积,但其数量略有升高。与睾丸假切除后诱癌的雄性大鼠相比,切除睾丸的大鼠经诱癌,有更低的GST-P阳性灶的面积;睾丸切除合并雌二醇处理,GST-P阳性灶的面积进一步降低。与仅化学诱癌的卵巢假切除雌性大鼠比,卵巢切除鼠诱癌后,GST-P阳性灶的面积稍有增加;对卵巢切除合用睾丸酮的大鼠诱癌,阳性灶的面积进一部增加。无论性腺切除与否,雄性大鼠比雌性大鼠有更高的GST-P表达。这些结果提示雌激素可抑制而雄激素则可促进化学诱癌大鼠肝中GST-P的表达。这一结果可能与临床上男性较女性易患肝癌有关。     

亚麻籽木酚素预防乳腺癌及与雌激素的关系

目的通过卵巢切除术建立雌性大鼠去势模型,探究亚麻籽粉木酚素预防乳腺癌的功能及与雌性激素的关系。方法将48只雌性Wistar大鼠随机分为基础饲料组（BD）、基础饲料去势组（BDC）、亚麻籽粉组（FS）和亚麻籽粉去势组（FSC）,每组12只,对全部大鼠进行二甲基苯蒽（DMBA）一次性灌胃（2mg/kg体重）建立诱发的乳腺癌实验动物模型;一周后对BDC组、FSC组大鼠行去势手术,连续观察21周,测定瘤体的体积和重量,并取乳腺组织进行病理学检查。结果实验期间动物一般状况良好,实验组大鼠未出现明显毒副作用;亚麻籽粉组（FS和FSC组）大鼠发生可触及肿瘤的时间较相应对照组晚2到4周;亚麻籽粉组大鼠单纯性增生和不典型增生以及乳腺癌发生率和病灶数均显著低于相应对照组（单纯性增生：FS vs BD,P=0.006＊＊;FSC vs BDC,P〈0.001＊＊;不典型增生：FS vs BD,P=0.048＊;FSC vs BDC,P=0.014＊;乳腺癌：FS vs BD,P=0.028＊;FSC vs BDC,P〈0.047＊）;亚麻籽粉组大鼠肿瘤体积和重量均小于基础饲料组;FS和FSC组研究结果提示亚麻籽粉木酚素抑制增生发生及肿瘤细胞的生长的能力与实验动物体内雌性激素水平有关（单纯性增生：P=0.008＊＊;不典型增生：P=0.042＊;乳腺癌：P=0.033＊）。结论亚麻籽粉木酚素可有效预防和降低化学诱癌剂DMBA所诱发的乳腺癌、癌前病变和单纯性增生的发生,预防乳腺癌的功能和效果受到体内雌性激素影响。本研究结果对未来实施木酚素预防乳腺癌及有效人群的筛选具有参考价值。     

不同剂量紫草素对雌激素加孕激素负荷法建立的子宫肌瘤大鼠的子宫组织ER, PR和p-ERK的蛋白表达影响

摘要 目的：探讨不同剂量紫草素对雌激素加孕激素负荷法建立的子宫肌瘤大鼠的子宫组织ER、PR和p-ERK的蛋白表达影响。方法：选择清洁级SD雌性大鼠60只,将其分为模型组、空白对照组、阳性对照组（雌二醇组）、低剂量紫草素组、中剂量紫草素组、高剂量紫草素组,每组10只大鼠。空白对照组、模型组大鼠给大鼠灌胃10 mL/kg?d生理盐水,阳性对照组给予戊酸雌二醇雌片灌胃,剂量为50 μg/kg?d,低剂量组灌胃给予5 mg/kg紫草素,中剂量组灌胃给予10 mg/kg紫草素,高剂量组灌胃给予20 mg/kg紫草素。记录大鼠的造模结果,观察大鼠的子宫形态、子宫病理组织学情况,对比各组大鼠的子宫重量、子宫系数、宫体最大直径、宫颈最大直径及平滑肌层厚度,检测每组大鼠的血清P、E₂水平及各组大鼠子宫组织ER、PR、p-ERK蛋白含量。结果：六组大鼠的子宫重量、子宫系数、宫体最大直径、宫颈最大直径及平滑肌层厚度指标两两比较,差异有统计学意义（P<0.05）;六组大鼠的炎性细胞浸润评分及子宫壁平滑肌厚度评分两两比较,差异有统计学意义（P<0.05）;六组大鼠的血清P、E₂水平两两比较,差异有统计学意义（P<0.05）;六组大鼠的子宫组织ER、PR、p-ERK蛋白水平两两比较,差异有统计学意义（P<0.05）。结论：紫草素具有抗子宫肌瘤活性的作用,随着紫草素剂量的增加,抗子宫肌瘤活性明显增加,可能与其可降低子宫组织ER、PR和p-ERK活性及血清P、E₂水平有关。     

HDAC1蛋白在乳腺癌中的表达及临床意义及与ER、PR的关系

目的：探讨乳腺癌组织中组蛋白去乙酰化酶1（HDAC1）的表达及临床意义。方法：应用免疫组织化学SP法检测78例乳腺癌组织和20例癌旁组织中HDAC1蛋白的表达情况并分析其与ER、PR之间的关系。结果：（1）HDAC1蛋白在78例乳腺癌中的阳性表达率为78.2%（61/78）,在癌旁组织中的阳性表达率为5%（1/20）,两组差异有统计学意义（P〈0.01）（2）乳腺癌组织中的HDAC1蛋白在ER或PR阴性乳腺癌组织中的表达分别高于其在ER或PR阳性乳腺癌组织中的表达（P〈0.01）结论：乳腺癌组织中的HDAC1蛋白过度表达与肿瘤发生发展密切相关。     

HDAC1蛋白在乳腺癌中的表达及临床意义及与ER、PR的关系

目的:探讨乳腺癌组织中组蛋白去乙酰化酶1(HDAC1)的表达及临床意义。方法:应用免疫组织化学SP法检测78例乳腺癌组织和20例癌旁组织中HDAC1蛋白的表达情况并分析其与ER、PR之间的关系。结果:(1)HDAC1蛋白在78例乳腺癌中的阳性表达率为78.2%(61/78),在癌旁组织中的阳性表达率为5%(1/20),两组差异有统计学意义(P<0.01)(2)乳腺癌组织中的HDAC1蛋白在ER或PR阴性乳腺癌组织中的表达分别高于其在ER或PR阳性乳腺癌组织中的表达(P<0.01)结论:乳腺癌组织中的HDAC1蛋白过度表达与肿瘤发生发展密切相关。     

DMBA诱导树鼩乳腺肿瘤(中缅树鼩)

目的:建立一个合适的乳腺癌动物模型将在研究人类乳腺癌的发生、发展、转移等方面中发挥着越来越重要的作用。7,12-二甲基苯并蒽(7,12-dimethylbenz anthracene,DMBA)在实验中能诱导大鼠产生乳腺肿瘤。树鼩的基因的结构与人类的相似程度比啮齿类动物要高,而且树鼩的自发性乳腺癌已经有被报道,因而树鼩很有可能是研究乳腺肿瘤更合适的动物模型。因此我们想用致癌剂DMBA诱导树鼩产生乳腺肿瘤而建立树鼩的乳腺肿瘤模型。方法:在这个研究中,我们采用了十只在分娩之后失去幼崽的雌树鼩,其中一半的树鼩在腰部双侧乳房的脂肪垫注射100 mg/kg的DMBA,其余的树鼩作为对照组没有作DMBA处理。对生成的肿瘤组织进行病理切片HE染色的形态特点分析以及免疫组化化学法测定Ki-67、雌激素受体、孕酮受体、人表皮生长因子受体-2、E-钙粘蛋白、P120连环蛋白的表达。结果:通过诊断在DMBA处理的树鼩中,5分之1发展浸润性导管癌,其余发展成原位导管癌。结果还证明了诱导出来的乳腺肿瘤的形态学和病理学特征与人类的浸润性导管癌相似。结论:结果显示我们采用DMBA注射失去幼崽的雌树鼩的乳腺来诱导乳腺肿瘤是有效的,诱导出来的肿瘤组织学特征与人的乳腺癌相似,诱导的肿瘤组织表达目前人常用的乳腺癌相关分子生物学标记,并且表达情况与人的乳腺癌相似。这表明了DMBA诱导树鼩乳腺癌可以提供一个适合于研究人类乳腺癌发生、发展、转移和治疗的动物模型。     

17β-雌二醇和跑台运动对去卵巢大鼠骨组织及子宫ERα蛋白表达影响的对比研究

目的：雌激素受体α（ERα）作为雌激素调节骨效应的主要受体在雌激素对骨量和骨代谢的调节中发挥重要的作用,为此本研究比较了17β-雌二醇（E2）和跑台运动对去卵巢大鼠骨组织和子宫ERα蛋白表达影响的异同。方法：将40只健康3月龄雌性SD大鼠,按体重分层后随机分为假手术、去卵巢、雌激素和运动四个组。手术1周后,雌激素组大鼠每周按体重颈部皮下注射三次17β-雌二醇,每次25μg／kg体重。运动组每周进行4次45min、速度18m／min、坡度5°的跑台训练。连续给药或运动处理14周后。采用放射免疫法和免疫组织化学法分别检测血清E2水平以及胫骨和子宫ERα蛋白表达的变化。结果：大鼠去卵巢后,子宫重量、子宫重量指数和血清E2水平显著下降,补充外源性17β-雌二醇后,三项指标均显著增加;但运动处理只能增加血清E2水平,对子宫重量和子宫重量指数均无显著影响。子宫EIh蛋白免疫组化结果显示：ERα在去卵巢组子宫内膜腔上皮、腺上皮及基质中的表达显著低于假手术组,而在雌激素组中的表达高于去卵巢组,运动组各部位ERα表达虽有所增加,但是增加的程度远低于雌激素组。胫骨近端ERα蛋白免疫组化结果显示：去卵巢后,胫骨近端骨骺端软骨细胞的细胞核内ERα表达减少,运动和雌激素干预后,胫骨近端ERα表达增加。结论：运动和雌激素处理均能刺激去卵巢大鼠子宫和骨组织ERα蛋白的表达,但运动处理无雌激素处理增加子宫重量的副作用。可见在绝经后骨质疏松的预防中,运动处理可能要优于雌激素处理。     

丹参酮Ⅱ A对人乳腺癌细胞株恶性表型逆转的研究

目的:研究丹参酮IIA体外诱导人乳腺癌细胞分化,逆转其恶性表型作用.方法:在体外培养的基础上,观察细胞形态学、细胞增殖动力学的变化.采用流式细胞仪的方法,定量检测了ER、nm23-1蛋白、抑癌基因c-fos、癌基因c-myc.结果:经无毒剂量的丹参酮IIA处理后,人乳腺癌细胞株MDA-MB-231形态趋向良性分化,细胞增殖指数明显降低,ER、nm23-l蛋白、抑癌基因c-fos明显升高,癌基因c-myc明显降低.结论:丹参酮IIA对人乳腺癌细胞株恶性表型有逆转作用,可能是通过抑制癌基因的表达,增加抑癌基因的表达,改变细胞形态结构和生物学特性.     

Estrogen receptor determination in fine needle aspirates of the breast. Correlation with histologic grade and comparison with biochemical analysis

Material obtained by fine needle aspiration (FNA) of 25 surgically removed breast carcinomas was tested for the immunocytochemical localization of estrogen receptor (ER) using the peroxidase-antiperoxidase method and a monoclonal antibody developed against human breast cancer ER. The results were compared to those obtained by the conventional biochemical analysis of cytosol protein. A semiquantitative relationship between the immunoperoxidase stain and the biochemical analysis suggests that cases in which greater than 70% of the cells stain and in which intense staining is present are likely to contain ER in a concentration of greater than 250 fmol/mg of cytosol. Less than 15% stained cells and an absence of intense staining is indicative of a concentration of less than 10 fmol/mg. In only one case was there a significant difference in positivity between the two methods, possibly as a result of a functional heterogeneity of the tumor cell population. Intense staining is strongly suggestive of a tumor of low histologic grade and was never seen in tumors with a high histologic grade or nuclear grade. The immunoperoxidase method of ER detection on material obtained by FNA is a semiquantitative means of selecting patients with breast cancer who are likely to respond to hormonal therapy. The method overcomes many important disadvantages of cytosol analysis and provides clinically significant information regarding the ER content and the degree of tumor differentiation.     

Prognostic value of pS2 protein and receptors for epidermal growth factor (EGF-R), Insulin-like growth factor-1 (IGF-1-R) and somatostatin (SS-R) in patients with breast and ovarian cancer

The prognostic value of EGF-R, IGF-1-R and SS-R, and of cytosolic estrogen-regulated pS2 protein, was studied in patients (pts) with primary breast and advanced ovarian cancer. Ovarian cancer tissues were negative for pS2 (by immunoradiometric assay) IGF-1-R and EGF-R contents (by ligand binding assay, LBA) were of no or moderate prognostic value for breast cancer pts (n = 214). For advanced ovarian cancer pts, EGF-R content determined by LBA (n = 55) showed no prognostic value, whereas EGF-R status (n = 55) determined by immunohistochemistry (MoAb 2E9) signiificantly correlated with progression of disease (P < 0.05). In breast cancer pts, both SS-R and pS2 showed no association with tumor size, nodal status and grade. For pS2 the best cut-off level with respect to relapse-free (RFS) and overall survival (OS) was found to be 11 ng/mg protein. Both SS-R (1 g% SS-R+, n = 135; P < 0.04) and pS2 (27% pS2+, n = 197; P < 0.001), which were mainly positive in ER+ tumors, were of prognostic value, especially within the subgroups with ER+/PgR+ tumors. Also within N+ and No pts the 5-yr RFS and OS showed a difference between pS2+ and pS2- (33 and 54% for N+, and 31 and 13% difference for No pts). In summary, SS-R and pS2 are valuable pronosticators in breast cancer pts, and prognostic significance of EGF-R in ovarian cancer pts needs further study.     

Demonstration of oestrogen receptor in symptomatic breast carcinoma, using fine needle aspiration cytology

Oestrogen receptor immunocytochemical assay (ER-ICA) was used to determine oestrogen receptor (ER) content of cells in fine needle aspirate (FNA) specimens from 88 breast carcinomas. In 49 of these the radioligand binding assay for oestradiol was available for comparison. The predictive value of ER-ICA staining for a positive radioligand binding assay (greater than 10 fmol/mg protein) was 95%. Although the predictive value of negative staining was only 66%, 34 out of 37 ER-ICA negative tumours had radioligand binding assays below 60 fmol/mg protein. ER-ICA staining showed a strong positive correlation with age of the patient, positivity being rare before the menopause. There was a weak inverse correlation with tumour grade but none with tumour size or lymph node status. The assessment of ER by immunocytochemistry using FNA cytology is a rapid technique, which may easily be repeated and provides a pre-operative assessment of ER status. It allows confirmation that tumour cells are present in the sample and an assessment of tumour heterogeneity.     

Epidermal growth factor receptors in human breast and endometrial carcinomas

The incidence and levels of epidermal growth factor receptor (EGFR) were studied in 67 breast tumors and 22 endometrial carcinomas. Estrogen receptors (ER) were also measured in all samples and progesterone receptors (PR) were analyzed in 57 breast samples and 21 endometrial tumors. A high level of EGFR expression is found in both breast and endometrial carcinomas, although the incidence of EGFR content is greater in breast carcinomas. 36% of breast tumors had EGFR at levels 3-49.5 fmol/mg membrane protein, whereas this percentage of positivity was 27% for endometrial tumors. In 51% of breast carcinoma and 73% of endometrial tumors, there was a positive ER content, whereas 53% of breast tumors and 62% of endometrial carcinomas were positive. A clear inverse relationship between EGFR content and ER and PR status has been observed in breast tumors. Our data confirm the previously described inverse correlation between expression of EGFR and estrogen receptors in human breast cancer. We also show here that there is a similar inverse relationship between EGFR content and ER levels in endometrial tumors.     

大白鼠妊娠早期(1—6天)子宫孕酮受体的变化

本实验中大鼠妊娠第三天(D_3)出现血浆孕酮含量和子宫细胞胞核中孕酮受体含量显著同步升高和胞质中孕酮受体含量明显下降的现象,为D_5胚泡着床准备了必要的条件。D_6时血浆孕酮,胞质和胞核中孕酮受体以及子宫重量均升高,标志胚泡着床后的生理变化。     

Comparative expression analysis of four breast cancer subtypes versus matched normal tissue from the same patients

Gene expression studies have been widely used in an effort to identify signatures that can predict clinical progression of cancer. In this study we focused instead on identifying gene expression differences between breast tumors and adjacent normal tissue, and between different subtypes of tumor classified by clinical marker status. We have collected a set of 20 breast cancer tissues, matched with the adjacent pathologically normal tissue from the same patient. The cancer samples representing each subtype of breast cancer identified by estrogen receptor ER(+/-) and Her2(+/-) status and divided into four subgroups (ER+/Her2+, ER+/Her2-, ER-/Her2+, and ER-/Her2-) were hybridized on Affymetrix HG-133 Plus 2.0 microarrays. By comparing cancer samples with their matched normal controls we have identified 3537 overall differentially expressed genes using data analysis methods from Bioconductor. When we looked at the genes in common of the four subgroups, we found 151 regulated genes, some of them encoding known targets for breast cancer treatment. Unique genes in the four subgroups instead suggested gene regulation dependent on the ER/Her2 markers selection. In conclusion, the results indicate that microarray studies using robust analysis of matched tumor and normal samples from the same patients can be used to identify genes differentially expressed in breast cancer tumor subtypes even when small numbers of samples are considered and can further elucidate molecular features of breast cancer.     

Significance of PIK3CA Mutations in Patients with Early Breast Cancer Treated with Adjuvant Chemotherapy: A Hellenic Cooperative Oncology Group (HeCOG) Study

Background

The PI3K-AKT pathway is frequently activated in breast cancer. PIK3CA mutations are most frequently found in the helical (exon 9) and kinase (exon 20) domains of this protein. The aim of the present study was to examine the role of different types of PIK3CA mutations in combination with molecular biomarkers related to PI3K-AKT signaling in patients with early breast cancer.

Methods

Tumor tissue samples from 1008 early breast cancer patients treated with adjuvant chemotherapy in two similar randomized trials of HeCOG were examined. Tumors were subtyped with immunohistochemistry (IHC) and FISH for ER, PgR, Ki67, HER2 and androgen receptor (AR). PIK3CA mutations were analyzed by Sanger sequencing (exon 20) and qPCR (exon 9) (Sanger/qPCR mutations). In 610 cases, next generation sequencing (NGS) PIK3CA mutation data were also available. PIK3CA mutations and PTEN protein expression (IHC) were analyzed in luminal tumors (ER and/or PgR positive), molecular apocrine carcinomas (MAC; ER/PgR negative / AR positive) and hormone receptor (ER/PgR/AR) negative tumors.

Results

PIK3CA mutations were detected in 235/1008 tumors (23%) with Sanger/qPCR and in 149/610 tumors (24%) with NGS. Concordance between the two methods was good with a Kappa coefficient of 0.76 (95% CI 0.69–0.82). Lobular histology, low tumor grade and luminal A tumors were associated with helical domain mutations (PIK3CAhel), while luminal B with kinase domain mutations (PIK3CAkin). The overall incidence of PIK3CA mutations was higher in luminal as compared to MAC and hormone receptor negative tumors (p = 0.004). Disease-free and overall survival did not significantly differ with respect to PIK3CA mutation presence and type. However, a statistically significant interaction between PIK3CA mutation status and PTEN low protein expression with regard to prognosis was identified.

Conclusions

The present study did not show any prognostic significance of specific PIK3CA mutations in a large group of predominantly lymph-node positive breast cancer women treated with adjuvant chemotherapy. Further analyses in larger cohorts are warranted to investigate possible differential effect of distinct PIK3CA mutations in small subgroups of patients.     

Mammographic density and hormone receptor expression in breast cancer: the Multiethnic Cohort Study

Background: It is unclear whether mammographic breast density, a strong risk factor for breast cancer, predicts subtypes of breast cancer defined by estrogen receptor (ER) and/or progesterone receptor (PR) expression. Methods: In a nested case–control study, we compared the breast density of 667 controls and 607 breast cancer cases among women of Caucasian, Japanese, and Native Hawaiian ancestry in the Hawaii component of the Multiethnic Cohort Study. A reader blinded to disease status performed computer assisted density assessment on prediagnostic mammograms. Receptor status was obtained from the statewide Hawaii Tumor Registry. Tumors were classified into ER+PR+ (n = 341), ER−PR− (n = 50), ER+PR−/ER−PR+ (n = 64), and unstaged/unknown (n = 152). Mean percent density values were computed for women with more than one mammogram. Polytomous logistic regression was used to estimate odds ratios (OR) and 95% confidence intervals (CI) while adjusting for confounders. Results: Mean percent density was significantly greater for ER+PR+ but not for ER−PR− tumors compared to controls after adjusting for age: 37.3%, 28.9% versus 29.4%, respectively. The overall OR per 10% increase in percent density were similar for ER+PR+ and ER+PR−/ER−PR+ tumors: 1.26 (95% CI 1.17–1.36) and 1.23 (95% CI 1.07–1.42), respectively. However, percent density was not found to be a predictor for ER−PR− tumors (OR 1.00, 95% CI 0.84–1.18). The results did not differ by ethnicity, nor by menopausal status, parity, or HRT use. Conclusions: Our findings indicate that within a multiethnic population, women with higher breast density have an increased risk for ER+PR+ but not ER−PR− tumors.     

Analysis of indoleamine 2-3 dioxygenase (IDO1) expression in breast cancer tissue by immunohistochemistry

Introduction

The immunosuppressive enzyme, indoleamine 2,3 dioxygenase (IDO), is overexpressed in many different tumor types including breast cancer. IDO inhibitors synergize with chemotherapy in breast cancer murine models. Characterizing IDO expression in breast cancer could define which patients receive IDO inhibitors. This study analyzed IDO protein expression in 203 breast cancer cases. The relationship between IDO, overall survival (OS), disease-specific survival (DSS), clinicopathologic, molecular, and immune tumor infiltrate factors was evaluated.

Methods

Expression of IDO, estrogen receptor (ER), progesterone receptor (PR), human epithelial receptor 2, cytokeratin 5/6, epithelial growth factor receptor, phosphorylated AKT, neoangiogenesis, nitrogen oxide synthetase 2 (NOS2), cyclooxygenase 2 (COX2), FoxP3, CD8, and CD11b on archival breast cancer tissue sections was evaluated by immunohistochemistry. Associations between IDO and these markers were explored by a univariate and multivariate analysis. Survival was analyzed using Kaplan–Meier (OS) and Wilcoxon two-sample (DSS) tests.

Results

IDO expression was higher in ER+ tumors compared to ER? tumors. IDO was lower in those with higher neoangiogenesis. OS was better in ER+ patients with high IDO expression. DSS was better in node-positive patients with high IDO expression. IDO activity positively correlates with NOS2. COX2 as positively correlated with IDO on univariate but not multivariate analysis. There was a trend toward greater numbers of CD11b+ cells in IDO-low tumors.

Conclusions

IDO protein expression is lower in ER- breast tumors with greater neoangiogenesis. Future clinical trials evaluating the synergy between IDO inhibitors and chemotherapy should take this finding into account and stratify for ER status in the trial design.