Immunohistochemical evaluation of estrogen and progesterone receptor content in 183 patients with endometrial carcinoma. Part II: Correlation between biochemical and immunohistochemical methods and survival.

One hundred eighty-three patients with endometrial carcinoma had both immunohistochemical and biochemical analysis of estrogen receptor (ER) and progesterone receptor (PR) content of the tumor. Biochemical analysis was done on a homogenate of uterine curettings; immunohistochemical stainings were done on formalin-fixed, paraffin-embedded sections. The biochemical method was scored quantitatively and the immunohistochemical, in a semiquantitative way. There was a significant correlation between the immunohistochemical categories of the malignant component of the tumor and the biochemical levels of receptor content (P = 0.0001). The sensitivity of the immunohistochemical method for the ER content was 78.5% and the specificity was 58.2%. The sensitivity for the PR was 52.5% and the specificity was 92.8%. Changing cut-off levels for positivity of the biochemical analysis changed the sensitivity and specificity. Survival was predicted by the immunohistochemical status of the ER (P = 0.001) and PR (P = 0.01). Similarly, it was predicted by the biochemical receptor status. Multivariate analysis of the immunohistochemical receptor status and the biochemical receptor status showed that the immunohistochemically determined estrogen status was the most significant predictor of survival (P = 0.001). The immunohistochemical analysis of sex steroid receptor status on formalin-fixed, paraffin-embedded tissue is not only possible and practical, but also predicts survival.     

PTEN缺失型子宫内膜癌中ER、PR的表达及意义

目的：探讨ER、PR在PTEN缺失型子宫内膜癌中的表达及PTEN、ER和PR表达特征与子宫内膜癌组织临床病理特征的关系。方法采用免疫组化EnVision法检测100例子宫内膜癌组织中PTEN、ER和PR表达。结果 PTEN在子宫内膜癌患者的缺失率为52．0%(52/100);PTEN缺失型子宫内膜癌组织中ER和PR阳性率分别为15．4%(8/52)和19．2%(10/52), PTEN高表达子宫内膜癌组织中ER和PR阳性率分别为72．7%(8/11)和63．6%(7/11);子宫内膜癌患者中PTEN-ER-PR-的比率为42．0%(42/100),均明显高于其它类型(P<0．05)。不同PTEN、ER和PR表达特征与子宫内膜癌患者组织学分级和病理分期有关(P<0．05),与肌层浸润无关。结论 PTEN、ER、PR三者联合检测可能对子宫内膜癌患者预后及治疗具有重要参考价值。     

Correlation of estrogen and progesterone receptors with histologic differentiation in endometrial adenocarcinoma.

Endometrial carcinomas from 58 patients were graded histologically, and the histologic grade was compared with the estrogen and progesterone receptor content of the tumor tissues. Receptor content was determined by multiple concentration saturation analysis and sucrose density gradient analysis. A positive correlation was found between the presence of estrogen and progesterone receptors and the degree of tumor differentiation. The majority (85%) of well-differentiated lesions contained significant amounts of both steroid receptor proteins. In contrast, only 13% of the poorly differentiated lesions were characterized by the presence of detectable levels of both estrogen and progesterone receptor proteins. The relationship of receptor content and degree of differentiation to prognosis and potential for response to hormonal therapy is discussed.     

Immunohistochemical detection of estrogen and progesterone receptor in human cornea

Objective: For treatment of postmenopausal keratoconjunctivitis sicca hormone therapy is favored by some clinicians. The likely morphological basis assessing the hormone receptor status in the human cornea has not been performed. Immunohistochemical staining methods provide the opportunity to evaluate the hormone receptor content within the histologic compartments of the cornea. The aim of our study was to assess and localize immunohistochemical hormone receptor staining in the human cornea. Methods: Formalin-fixed and paraffin-embedded specimens of three pre- and three postmenopausal women were assessed for localization of estrogen receptor (ER) and progesterone receptor (PR) expression with established immunohistochemical hormone receptor staining methods. Results: No nuclear staining reaction was found in the epi- and endothelial layers of the corneas. The stroma of the corneas showed no immunohistochemical staining reaction in all cases. We found cytoplasmatic PR staining of the endothelial layer in two cases. Conclusions: We found no morphological basis in the human cornea for the use of topical steroid hormone treatment in postmenopausal keratoconjunctivitis sicca. Hormone receptor expression in the conjunctiva or in the lacrimal gland may have an impact in some patients showing relief of symptoms in postmenopausal dry eye syndrome.     

子宫内膜组织ER、PR及PCNA表达与子宫内膜增生过长的关系

目的：探讨子宫内膜增生过长的发病机理，为临床内分泌治疗提供理论基础。方法：应用免疫组织化学Ｓ－Ｐ法，对手术切除和诊刮的６７例不同时期子宫内膜和不同类型增生过长子宫内膜标本进行ＥＲ、ＰＲ和ＰＣＮＡ含量检测分析。结果：ＥＲ、ＰＲ在正常增生期子宫内膜中的含量显著高于分泌期（Ｐ＜０．０１），在单纯性增生过长和复杂性增生过长子宫内膜中ＥＲ、ＰＲ的含量也有显著差异（Ｐ＜０．０１）。各组增生过长子宫内膜中ＥＲ的含量高于ＰＲ（Ｐ＜０．０１）。ＰＣＮＡ在内膜分泌期和伴有非典型增生的子宫内膜中含量高（Ｐ＜０．０１）。结论：ＥＲ主要与子宫内膜增生过长有关，ＰＣＮＡ在非典型增生子宫内膜中过表达可能与宫内膜异常生长有关。     

Immunohistochemical analysis of estrogen receptor alpha, estrogen receptor beta and progesterone receptor in normal human endometrium

The endometrium expresses estrogen (ER) and progesterone receptors (PR), which are involved in autocrine and paracrine regulation processes in response to estrogen and progesterone. The aim of the present study was to evaluate immunohistochemical distribution patterns of estrogen receptor alpha (ER alpha), estrogen receptor beta (ER beta) and PR in normal human endometrial tissue with the use of monoclonal antibodies. Human endometria were obtained from 17 premenopausal patients undergoing surgery for non-malignant diseases and were classified to be in proliferative, early secretory and late secretory phases by histological and anamnestical means. Distribution patterns of the steroid receptors were evaluated using the IRS-score and the Mann-Whitney rank-sum test was used to compare the means. Correlation was assessed with the Spearman factor and linear regression analysis. ER alpha and PR expression decreased significantly (p<0.05) in glandular epithelium from the proliferative to the late secretory phase. ER beta expression showed a similar significant decrease (p<0.05), although staining intensity was lower than that of ER alpha. A significant correlation between expression of all three steroid receptors was observed (p<0.001). Distribution patterns of ER alpha, ER beta and PR in normal human endometrium showed a cyclic variation during the menstrual cycle. A significant correlation between expression of ER alpha, ER beta and PR was also demonstrated using regression analysis, indicating dependence of expression of these three steroid receptors. The present study shows the presence of steroid receptors in human endometrial epithelium, indicating that these cells respond to estrogen and progesterone and thus playing a significant role in endometrial physiology.     

Immunohistochemical expression of estrogen and progesterone receptors in endometrial polyps and adjacent endometrium in postmenopausal women

OBJECTIVE: To detect the presence of estrogen (ER) and progesterone (PR) receptors in endometrial polyps and adjacent endometrium in postmenopausal women. METHODS: Forty-four consecutively enrolled postmenopausal patients were submitted to operative hysteroscopy. These patients had diagnosed benign endometrial polyp. The presence of ER and PR was determined in endometrial samples and polyps by immunohistochemical method and the slides were evaluated using a semiquantitative analysis. RESULTS: In the glandular epithelium, the median of the ER score was 7.0 in the polyps and 5.0 in the endometrium (P<0.0001) and the median of the PR was 6.0 in the polyps and 4.0 in the endometrium (P<0.0001). In the stroma, the median of the ER score was 6.0 in the polyps and 5.0 in the endometrium (P=0.021) and the median of the PR score was 4.0 in the polyps and 4.5 in the endometrium (P=0.34 ). CONCLUSIONS: Our data suggests that steroids receptors present a crucial role in the phisiopathology of the endometrial polyps in postmenopausal women, specially the estrogen receptors.     

Immunohistochemical evaluation of human epidermal growth factor receptor 2 and estrogen and progesterone receptors in invasive breast cancer in women

Introduction

Estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER-2) expression are crucial in the biology of breast carcinoma. HER-2/neu gene is amplified and overexpressed in 15-30% of invasive breast cancers. HER-2-positive breast cancers have worse prognosis than HER-2 negative tumors and possess distinctive clinical features. The aim of this study was to assess the expression of HER2 in cancer tissue of patients with invasive breast cancer in correlation with tumor type, histological grade, tumor size, lymph node status, and expression of estrogen receptor and progesterone receptor.

Material and methods

Formalin-fixed, paraffin-embedded tissues from 40 patients with invasive HER-2-positive breast cancer and from 191 patients with HER-2-negative breast cancer were used in this study. HER2 expression was determined using the test HerceptTest™ DAKO.

Results

Among 231 cases of breast cancer, 18 invasive lobular carcinomas and 213 invasive ductal carcinomas were diagnosed. Sixty percent of HER-2-positive breast cancers were ER-positive compared with 77% in the HER-2-negative group (p = 0.002). The expression of PR was observed in 43% of HER-2-positive breast cancers and in 72% of HER2-negative tumors (p = 0.003). Excessive expression of HER2 protein was detected in 60% of patients positive for estrogen receptors, which may worsen prognosis in these patients.

Conclusions

Determination of HER2 overexpression in breast cancer patients, allows for a determination of a group of patients with a worse prognosis.     

Association of exogenous estrogen and endometrial carcinoma.

To determine the association between the incidence of endometrial cancer and the use of estrogen in menopausal and post-menopausal women, we retrospectively compared 317 patients with adenocarcinoma of the endometrium with an equal number of matched controls having other gynecologic neoplasms; 152 patients used estrogen, as compared to 54 of 317 controls. Thus, the risk of endometrial cancer was 4.5 times greater among women exposed to estrogen therapy. When estrogen use was adjusted for concomitant variables such as obesity, hypertension, diabetes, parity, referral pattern, age at diagnosis, year of diagnosis and other gynecologic neoplasms, the magnitude of the increased relative risk was associated with several of these variables, and was highest in patients without obesity and hypertension. Exogenous estrogen therapy is associated with an increased risk of endometrial carcinoma, but this increased relative risk is less apparent in patients with physiologic characteristics previously associated with an increased risk.     

Incorporation of perforated and demineralized cortical bone allografts. Part I: radiographic and histologic evaluation

Massive cortical bone allografts have been found to incorporate slowly into host bone. They are subject to complications such as nonunion, fatigue fracture and infection. In an attempt to improve osteoinduction in cortical bone allografts, laser perforated and partially demineralized cortical bone allografts were orthotopically transplanted into the sheep tibia. In this model, mid-shaft tibial bone allografts from out-bred sheep donor animals were prepared by partial demineralization and drilling of 0.33-mm diameter holes with a pulsed, 2.94-microm wavelength Erbium:Yttrium-Aluminum-Garnet laser. Recipient animals of the same out-bred strain were divided into three groups of eight according to the type of cortical allograft used: group 1, fresh-frozen, no treatment; group 2, laser hole grid; and group 3, laser hole grid and partial demineralization. Plain films were taken in two standard views at monthly intervals. Incorporation was evaluated at nine months postoperatively. Longitudinal radiographic data was correlated to a histologic and morphometric evaluation of each bone graft. Computer tomography was used for the latter analysis. Results showed that untreated allografts, although surrounded by a periosteal bone cuff, were poorly incorporated. Partial demineralization lead to excessive resorption of allografts, but little new bone formation. Laser perforation and partial demineralization induced complete incorporation of allografts into the host bone. Based on the results of the radiographic, histologic and morphometric evaluation, the development of laser-perforated and partially demineralized bone allografts was proposed for clinical use.     

Subcellular distribution of estrogen receptor and progesterone receptor with and without specific ligand.

The estrogen receptor (ER) and progesterone receptor (PR) content of cultured human breast carcinoma cells (MCF-7) was determined by biochemical assay, immunoblot analysis, and immunohistochemical assay under varying conditions of hormonal stimulation. The ER and PR content in cytosolic and nuclear extracts varied with steroid treatment. However, both the amount and distribution of each receptor in these extracts was virtually the same when determined by steroid binding and immunoblot analyses. Two immunocytochemical parameters (staining intensity and proportion of cells stained) correlated with the quantitative analyses of ER and PR, but not with the subcellular distribution. When MCF-7 cells were grown for 4 days in charcoal-stripped serum without phenol red, 93% of total ER was found in the cytosol (10 mM KCl), whereas short-term treatment with 5 nM estradiol resulted in the appearance of 82% of total ER in the nuclear extract (400 mM KCl). With either cell treatment only nuclear staining for ER was observed. Progesterone receptor was virtually undetectable in the same cells by any method. After 4 days of treatment by 5 nM estradiol, PR was strongly induced (50-fold) in MCF-7 cells as determined by all three methods. As observed for ER, 95% of total induced PR was found in the cytosol in the absence of a progestin. Short-term treatment with 5 nM ORG 2058, a synthetic progestin, resulted in the appearance of 42% of total PR in the nuclear extract. However, only strong nuclear staining for PR was observed in either the presence or absence of a progestin. These findings are consistent with the current view of ER and PR as nuclear receptors present in at least two forms. One of these, the unoccupied form of the receptor, is easily removed from the nucleus by hypotonic buffers during the cell homogenization process and appears in the cytosolic extract. The other form of the receptor, the steroid-occupied form, is more tightly bound to nuclear components and is removed from nuclei only under more vigorous extraction conditions.     

雄激素受体在乳腺癌中的表达及其与雌、孕激素受体和HER2的关系

目的 研究雄激素受体(AR)在乳腺浸润性导管癌中的表达及其与雌激素受体(ER)、孕激素受体(PR)和HER2状态的关系,探讨其作为乳腺癌治疗靶点的可行性.方法 采用免疫组织化学EnVision法检测AR、ER、PR、HER2在175例乳腺浸润性导管癌中的表达,依据结果分为腺腔A型、腺腔B型、HER2过表达型和三阴性型(ER-/PR-/HER2-)组.结果 175例中AR阳性88例(50.3%),AR表达与ER、PR、HER2均呈正相关(P＜0.01).腺腔A型53例(30.3%),腺腔B型33例(18.9%),HER2过表达型23例(13.1%),三阴性型66例(37.7%),AR阳性率分别为56.6%(30/53),75.8%(25/33)、47.8%(11/23)和33.3%(22/66),组间AR阳性率差异显著(x2=17.054,P=0.001).三阴性型组AR阳性者核分裂象较少(x2=5.140,P=0.023),腺腔A型组AR阳性者多为年轻患者(x2=4.567,P=0.033),差异有统计学意义.其他组内AR表达与否和临床病理学特征比较无统计学意义.结论 AR在乳腺癌中有较高的阳性率,可作为乳腺癌,特别是三阴性型乳腺癌的治疗靶点.     

雄激素受体在乳腺癌中的表达及其与雌、孕激素受体和HER2的关系

目的 研究雄激素受体(AR)在乳腺浸润性导管癌中的表达及其与雌激素受体(ER)、孕激素受体(PR)和HER2状态的关系,探讨其作为乳腺癌治疗靶点的可行性.方法 采用免疫组织化学EnVision法检测AR、ER、PR、HER2在175例乳腺浸润性导管癌中的表达,依据结果分为腺腔A型、腺腔B型、HER2过表达型和三阴性型(ER-/PR-/HER2-)组.结果 175例中AR阳性88例(50.3%),AR表达与ER、PR、HER2均呈正相关(P＜0.01).腺腔A型53例(30.3%),腺腔B型33例(18.9%),HER2过表达型23例(13.1%),三阴性型66例(37.7%),AR阳性率分别为56.6%(30/53),75.8%(25/33)、47.8%(11/23)和33.3%(22/66),组间AR阳性率差异显著(x2=17.054,P=0.001).三阴性型组AR阳性者核分裂象较少(x2=5.140,P=0.023),腺腔A型组AR阳性者多为年轻患者(x2=4.567,P=0.033),差异有统计学意义.其他组内AR表达与否和临床病理学特征比较无统计学意义.结论 AR在乳腺癌中有较高的阳性率,可作为乳腺癌,特别是三阴性型乳腺癌的治疗靶点.     

Aberrations in the progesterone receptor gene and the risk of recurrent endometrial carcinoma

A case-control study was performed in order to determine whether expression of the progesterone receptor (PR) and/or aberrations of the PR gene contribute to the development of recurrent endometrial carcinoma. Primary tumours from 44 patients with recurrence of stage I endometrial carcinoma (patients) within 3 years after initial treatment were compared with tumours from 44 matched patients who were free of recurrence for a minimum of 3 years (controls). Paraffin wax-embedded primary tumours (n = 88) and recurrent tumours (n = 32) were analysed immunohistochemically for PR expression. A staining index (SI = 0-9) based on the staining intensity and the number of stained cells was calculated. DNA extracted from paraffin wax-embedded tissues was subjected to PCR-restriction fragment length polymorphism analysis (PCR-RFLP) for determination of the PROGINS DNA sequence alterations and the +331G/A-promoter polymorphism. Low PR expression (SI < 1.0) was observed in 7% of primary tumours derived from controls, 25% of primary tumours from patients with recurrence, and 38% of recurrent tumours. The expression of PR was significantly lower in primary tumours from patients with recurrence (SI = 4.0 +/- 0.5) than in the tumours in the control group (SI = 5.6 +/- 0.5) (T-test for paired analysis, p < 0.05). The PROGINS and +331G/A-promoter polymorphism were not related to age at diagnosis, tumour grade or myometrial invasion. The +331G/A-promoter polymorphism was present in 14% of primary tumours from patients without recurrence, compared with 17% of patients with recurrence. The PROGINS polymorphism was observed in 16% of primary tumours from patients without, and in 34% of patients with, recurrence (OR 2.6; 95% CI: 0.9-7.6). Most interestingly, patients who carried the PROGINS variant and in whom a PR-expressing tumour was diagnosed were at significantly enhanced risk of relapse (OR 4.7; 95% CI: 1.3-17.1). In conclusion, low PR expression tended to be associated with recurrent disease, and PR expression in tumours from patients carrying the PROGINS allele was predictive of the risk of recurrence.     

Immunohistochemical detection of the estrogen receptor-alpha and progesterone receptor in the canine pregnant uterus and placental labyrinth

The presence of hormone receptors is as important as the amount of hormone to predict hormone action. Therefore, the presence of estrogen receptors of the alpha subtype (ER-alpha) and progesterone receptors (PR) was evaluated in six pregnant uteri including the placenta and in three postpartum uteri of dogs. This preliminary study is part of our immunohistochemical research project on steroid hormone receptor distribution in the canine female genital tract. Specific staining for ER-alpha or PR was found only in cell nuclei. Staining for ER-alpha was rare in the various cell types of pregnant and postpartum uteri. Staining for PR was absent or weak in epithelial cells. Moderate staining for PR was observed in endometrial stromal cells and myometrial smooth muscle cells, two cell types playing an important role in the maintenance of pregnancy. Stromal cells stained more frequently positive for ER-alpha and PR than epithelial cells, indicating that both hormones may act on epithelial cells indirectly via stromal cells. In the placental labyrinth, fetal cells showed no evidence of ER-alpha or PR. In contrast, both receptors were present in maternal mesenchymal cells that were located around the basement membrane of the maternal blood vessels. These cells showed signs of decidualization. No difference in PR distribution was seen between pregnant and postpartum uterine tissue, suggesting that during parturition the decrease in serum progesterone levels and the concomitant increase in the estrogen/progesterone ratio are probably more important than the decline in receptor availability.     

PTEN expression in breast and endometrial cancer: correlations with steroid hormone receptor status.

OBJECTIVE: The PTEN (MMAC1/TEP1) tumor suppressor gene is frequently mutated and homozygously deleted in human neoplasms, but there is only sparse information about PTEN protein expression in hormone-dependent female tumors. Therefore, we investigated PTEN expression in 68 breast and 43 endometrial carcinomas. METHODS: For PTEN protein detection, we used Western blot analysis followed by densitometry and compared these data with clinicopathologic parameters, the estrogen receptor (ER) and progesterone receptor (PR) status, HER2/neu and the proliferation marker Ki67. RESULTS: We were able to show significantly decreased PTEN protein expression in endometrial carcinomas compared with normal endometrial tissue samples, especially in the endometrioid histological subtype. In contrast, PTEN downregulation was found more rarely in breast cancer. Lower PTEN expression in breast cancer correlated significantly with high ER immunoreactivity (p = 0.008) and was weakly associated with PR expression (p = 0.055) and low histological grading (p = 0.081). No correlation with any of these parameters was observed in endometrial tumors. In both tumor types, no association of PTEN expression with any other analyzed parameter was found. CONCLUSIONS: These results suggest that PTEN expression plays different roles in the pathogenesis of endometrial carcinomas and breast cancer. In mammary carcinomas, loss of PTEN expression is mainly found in more differentiated tumors and is probably not a major event in carcinogenesis.