Plasminogen activator activity in human prostate and breast tumors: relationship to steroid receptors

Using a fluorometric assay based on the activation of human plasminogen, plasminogen activator (PA) activity was measured in cytosolic and lysosomal extracts prepared from normal, benign hyperplastic (BPH), and carcinomatous human prostates. In all three types of prostatic tissue, the lysosomal extracts had much higher concentrations of PA activity than did the cytosolic extracts. The mean PA activity in lysosomal extracts of the carcinomatous prostates was 170% and 85% higher than that measured in normal and BPH prostates, respectively (Student's t-test, p less than .05). With prostatic carcinoma and BPH specimens there was an inverse (negative) relationship between lysosomal PA activity and the nuclear concentration of androgen-receptors (correlation coefficient, -0.84). By comparison in specimens of human breast tumors, there were weakly positive correlations between PA activity and either estrogen (ER) or progestin (PR) receptors (correlation coefficients of + 0.23 and + 0.54, respectively). While as a group ER+, PR+ breast tumors had higher PA activity that ER+, PR- or ER-, PR- tumors, the differences were not statistically significant (Student's t-test, p greater than .05). Thus in breast tumors, it is uncertain whether high levels of PA activity are indicative of hormonal dependence. However, our findings with prostatic tumors infer that in contrast, high concentrations of this enzyme may reflect a malignant phenotype characterized by a decrease in both androgen responsiveness and differentiation.     

Steroid receptor profile and receptor stability in subfractions of human prostatic tissues

Summary Androgen (AR), progesterone (PR), and estrogen (ER) receptor contents in cytosol and salt-extractable nuclear subcompartments from 6 normal, 39 benign hyperplastic (BPH), and 7 malignant prostatic tissue specimens were analyzed by radioligand-binding assay techniques. In addition, the temperature stability of AR and PR was measured in another three BPH specimens. Five punch-needle biopsy samples from prostate cancers were also analyzed for AR and PR content. All receptor data were calculated from saturation analyses. The highest AR content was found in the cytosol and nucleic from malignant prostatic tissues. The highest PR concentrations were found in BPH cytosol, whereas nuclei of all types of tissues were negative with regard to this receptor. Markedly lower concentrations of ER were found in cytosol and nuclei from BPH as compared with malignant and normal tissues. PR was the most temperature-stable receptor; a marked receptor loss at room temperature was not registered until after 12 h. AR was stable for 4–5 h in cytosol and for 8–9 h in nuclei. Needle-biopsy specimens from prostate cancer showed highly variable and confusing results for AR and PR content, indicating that microassay studies using biochemical techniques on small tissue samples are unreliable and should not be recommended.     

Immunocytochemical localization of estrogen receptors in spontaneous and experimentally induced canine benign prostatic hyperplasia

Estrogens are believed to play a critical role in the etiology of canine benign prostatic hyperplasia (BPH); however, the mechanism has not been elucidated. To gain insight into this problem, we investigated the immunocytochemical localization of estrogen receptors (ER) in normal prostates, spontaneous BPH, and experimentally induced BPH by using a monoclonal ER antibody (H222). In all canine prostates the majority of ER was localized in nuclei of the same histological components: (1) transitional epithelium and subjacent stroma of the prostatic urethra, (2) periurethral prostatic ductal epithelium, and (3) prostatic stroma. ER content in the stroma was highest in the periurethral region of the prostate. Among the different groups of dogs, differences in ER location were seen only in the glandular epithelium. No ER was found in the glandular epithelium of normal prostates of young untreated dogs. In striking contrast, glandular epithelium of spontaneous BPH contained specific nuclear ER staining, though this staining was heterogeneous and was observed in only a minority (less than 10%) of the acinar epithelial cells. ER-positive acini in BPH were located predominantly in the periurethral region. These data demonstrate anatomical and biochemical heterogeneity of prostatic components and indicate that the estrogen sensitivity of prostatic cells is heterogeneous. If estrogen does play a role in BPH, it appears to act selectively rather than uniformly throughout the prostate. We reasoned that if glandular epithelial ER are involved in the development of spontaneous BPH, one might expect to find the same location of ER in BPH that was induced experimentally by specific types of treatment with androgens +/- estradiol. However, among hormone-treated dogs the presence of ER-positive prostatic glandular epithelium varied with the type of hormonal treatment but did not correlate with the experimental induction of glandular BPH. Some treatment groups with induced BPH had ER-positive prostatic glandular epithelial nuclei (with the same extent and pattern of ER localization as in spontaneous BPH); however, other treatment groups with induced BPH had ER-negative glandular epithelium. These data indicate either that glandular epithelial ER may not be involved in the pathogenesis of canine BPH or that there may be different types of BPH that have different etiologies. Possible mechanisms by which estrogen may affect the canine prostate are discussed in light of these new data on ER location.     

前列腺癌中E-钙粘素、p16及雌激素受体基因甲基化的检测及其临床意义

目的:研究前列腺病变中E-钙粘素、p16和雌激素受体(ER)基因启动子CpG岛甲基化状况及其与临床资料的关系,探讨基因过甲基化在前列腺癌(PCa)发生发展中的作用及其临床意义。方法:收集石蜡包埋前列腺全切术标本:良性前列腺增生(BPH)13例,高级别前列腺上皮内瘤(HGPIN)10例,有随访记录的PCa 20例。采用甲基化特异性PCR(MSP)技术对E-钙粘素、p16和ER基因CpG岛甲基化进行检测。结果:E-钙粘素、p16和ER基因过甲基化在PCa中的发生率分别为30%、25%和65%。非恶性病变(BPH和HGPIN)很少发生DNA过甲基化。结论:PCa的发生及进展与E-钙粘素、p16和ER基因过甲基化密切相关,检测甲基化状况对HGPIN与PCa的鉴别诊断可能有辅助作用。E-钙粘素和ER基因过甲基化可能提示预后差。     

小儿血管瘤与血管畸形组织中性激素受体的表达

目的 了解小儿血管瘤与血管畸形组织中的雌激素受体（ＥＲ）,孕激素受体（ＰＲ）和雄激素受体（ＡＲ）水平,并比较其差异,方法 对３１例血管瘤和３６例血管畸形患儿病变组织标本进行ＨＥ染色和酶联组化法复染。光镜下检测ＥＲ,ＰＲ和ＡＲ。结果 血管瘤与血管畸形组织中均含有上述３种受体,但血管瘤中ＥＲ,ＰＲ和ＡＲ水平明显高于血管畸形组（Ｐ〈０．０１）,而且增生期血管瘤组织中ＥＲ,ＰＲ和ＡＲ高于消退期血管瘤组织（     

Prostatic cancer/benign prostatic hypertrophy. Subcellular distribution of estradiol/androgen receptors

Six microsomal population of estradiol and androgen receptors have been characterized in human benign prostatic hypertrophy (BPH) and prostate cancer (PCa). Estradiol receptor (ER) and androgen receptors (AR) were extracted using 0.6 M KCL and determined by the dextran-coated charcoal method. ER and AR levels were smaller in BPH plasma membranes (PM) than in Pca cases. For functions 3, 4, 6, the ER values in PCa were 25-38% less with regard to BPH ER values. Whereas in PCa, AR values obtained in all fractions were higher when compared to BPH AR values. In benign prostatic hypertrophy and prostatic cancer, ER and AR levels were significantly higher in the nuclear fraction. In the nuclear fraction, ER and AR levels in BPH and PCa were significantly different. The subcellular distribution of AR and ER in BPH and PCa constitutes a reservation mechanism and processing a receptors for their continued growth.     

The relationship of estrogen receptor status to DNA ploidy in breast cancer

The relationship of estrogen receptor(ER) status to DNA ploidy was investigated in 121 patients with breast cancer who underwent surgery. Lymph node status was evaluated histologically and ER levels were determined by the dextran-coated charcoal method, with a level of 3 fmol/mg·protein being considered positive. Flow cytometric DNA content was analyzed using paraffin-embedded tissue blocks. Sixty-three per cent of the specimens were ER+, while 37 per cent were negative. Sixty-one patients (50.4 per cent) were diploid and 60 aneuploid. A statistically significant correlation between the ER positivity rate and diploid DNA was found. Higher ER levels were seen in the postmenopausal patients with diploid tumors than in those with aneuploid tumors and there was a significant tendency for ER levels to be higher in the diploid tumors. Nodal status was not correlated with ER positivity or ploidy pattern. The present results indicate that ER levels are correlated with DNA ploidy, and reflect the degree of functional differentiation.     

Influence of age and endocrine factors on the volume of benign prostatic hyperplasia.

To determine whether endocrine factors influence the volume of benign prostatic hyperplasia (BPH), 23 hormonal factors were measured in the serum of 64 men ages 42 to 71 years with low volume prostatic cancer and these levels were correlated with the volume of benign hyperplastic tissue in their radical prostatectomy specimens. With age there was a significant increase in the volume of BPH. Also with age there was a significant decrease in the serum levels of free testosterone, androstenedione, dehydroepiandronsterone (DHA), dehydroepiandronsterone sulphate (DHA-S), delta 5-androstenediol, and 17-hydroxypregnenolone, and a significant increase in sex hormone-binding globulin (SHBG), LH, and FSH. When BPH volume and hormone levels were corrected for age, BPH volume correlated positively with free testosterone, estradiol, and estriol. These data indicate that with age patients with larger volumes of BPH have higher serum androgen and estrogen levels suggesting that serum androgen and estrogen levels may be factors in the persistent stimulation of BPH with age. If so, therapeutic attempts at lowering plasma testosterone levels, reducing estrogen levels, or blocking androgenic stimulation through other mechanisms may interfere with the progression of BPH with age. Conversely, the fact that androgen production declines gradually with age may explain the observation that only 20 to 30% of men who live to age 80 require surgical treatment for urinary obstruction from BPH.     

前列腺癌和癌旁组织中雌激素受体α和β的表达及意义

目的 通过测定前列腺癌中癌组织、癌旁组织和良性前列腺增生组织中雌激素受体(estrogen receptor,ER)α和β的表达水平,探讨ER在前列腺腺癌发生、发展中的变化和作用机制.方法 采用超高敏链酶亲合素-过氧化物酶法检测28例前列腺腺癌标本中癌和癌旁组织以及29例良性前列腺增生组织中ERα和ERβ的表达水平,对比其在各组之间表达的差异,分析ERα和ERβ的表达水平与前列腺癌患者的Gleason评分、临床分期、年龄和TPSA的关系.结果 前列腺癌组织中ERα主要在间质细胞表达.ERα在前列腺癌组织、癌旁组织和良性前列腺增生组织上皮细胞的表达阳性率分别为0%、14%、24%,间质细胞阳性率分别为57%、68%、31%,组间比较差异有统计学意义(P＜0.05).前列腺癌上皮细胞和间质细胞均有ERβ的表达且差异无统计学意义(P＞0.05).ERβ在前列腺癌组织、癌旁组织和良性前列腺增生组织上皮细胞的表达阳性率分别为39%、64%、29%;间质细胞阳性率分别为50%、75%、79%,组间比较差异有统计学意义(P＜0.05).ERβ在不同Gleason评分前列腺癌组织中表达差异有统计学意义(P＜0.05).结论 前列腺癌中ERα主要在间质细胞表达;ERβ在上皮细胞和间质细胞均有表达.ERα和ERβ的表达水平在前列腺癌组织、癌旁组织和良性前列腺增生组织均存在差异.ER口与前列腺癌变发生和恶性程度相关.

Abstract：

Objective To investigate the expression of estrogen receptor (ER) α and β in human prostate cancer (PC), peri-cancer tissue and benign prostatic hyperplasia (BPH) tissue, and to discuss the role of estrogen receptor in prostate cancer. Methods The expression of ERα and ERβ in PC (n=28), peri-cancer tissue (n=28) and BPH (n=29) were detected by immunohistochemistry with En vision method. The ERα and ERβ expression were compared among different tissues by chisquare. The relationship between ER expression and related clinicopathologic features was statistically analyzed by spearman rank collection. Results ERα was localized dominantly in the stromal cell of PC. There were significant differences of the expression of ERα in PC, peri-cancer tissue and BPH tissue (epithelial cell 0%, 14%, 24%, P＜0. 05; stromal cell 57%, 68%, 31%,P＜0. 05). ERβ was localized in both epithelial and stromal cell of PC. There were significant differences of the expression of ERβ in PC, peri-cancer tissue and BPH tissue (epithelial cell 39%, 64%, 29%, P＜0.01; stromal cell 50%, 75%, 79%, P＜0.05). There was a significant difference of the expression of ERβ in different Gleason scores of PC tissue. Conclusions ERα is localized in the stromal cell of PC tissue.ERβ is localized in both epithelial and stromal cell of PC tissue. The ERβ might be related to the tumor differentiation of PC.     

Quantitation of cytosolic and nuclear estrogen and progesterone receptor in benign, untreated, and treated malignant human prostatic tissue by radioligand binding and enzyme-immunoassays.

Estrogen receptor (ER) and progesterone receptor (PgR) were quantitated in benign and malignant human prostatic tissue by using radioligand binding assays (RBA) and enzyme-immunoassays (EIA). Using a hydroxylapatite exchange method for ER, little or no nuclear ER (ERN) could be detected, but with the EIA both cytosolic (ERC) and ERN were detected in almost all specimens, although in meager concentrations. Tissue from patients with carcinoma had significantly higher ERC concentrations than tissue from patients with benign disease. Specimens from estrogen-treated patients had significantly lower ERC:ERN ratios than those from untreated patients. Progesterone receptor was detected in virtually all specimens by both methods, at concentrations higher than those of ER. Carcinoma tissue with a high malignant involvement contained significantly less PgR than benign tissue. Using either method, the highest concentrations of PgR were observed in tissue from carcinoma patients treated with estrogen. Overall, the data suggest that although human prostatic tissue contains only modest amounts of ER, this is active in that treatment with estrogen promotes association of this receptor with the nuclear fraction and increases PgR content.     

Histological localization of estrogen receptors in normal and diseased human prostates by immunocytochemistry.

The role of estrogens and estrogen receptors (ER) in the human prostate remains unresolved. In this study we have used the monoclonal ER antibody H222 to investigate the histological localization of ER in normal and diseased human prostates by immunocytochemistry. Prostate tissue was obtained from 3 young organ donors (Group I-normal prostate), from 14 prostates removed by radical prostatectomy or radical cystoprostatectomy, which had caused no or only mild obstructive symptoms (Group II-non-obstructive prostate), and from 11 prostates removed by suprapubic prostatectomy, which had caused severe obstructive symptoms due to a large benign prostatic hyperplasia (BPH) (Group III-obstructive prostate). In prostates of all groups ER were found to be in nuclei of the prostatic urethra and of the periurethral prostatic duct. In striking contrast, ER in the interglandular prostatic stroma was not as homogeneous among the different groups. We observed a low concentration of ER in the stroma of normal prostates, the highest concentration in non-malignant stroma of non-obstructive prostates, and no ER at all in stroma of obstructive prostates. Based on the immunocytochemical localization of ER in normal and diseased human prostate, our results indicate that stromal growth in obstructive BPH may not be mediated via ER. However, we cannot exclude that an increase of stromal ER concentration (as observed in non-obstructive prostates) is directly involved in induction of BPH, leading further prostate growth thereafter into an estrogen independent state.     

Evidence that Serenoa repens extract displays an antiestrogenic activity in prostatic tissue of benign prostatic hypertrophy patients.

A double-blind placebo-controlled study was performed in 35 benign prostatic hypertrophy (BPH) patients never treated before. The patients were randomized into two groups, the 1st (18 cases) receiving Serenoa repens extract (160 mg t.d.) for 3 months up to the day before the operation of transvesical adenomectomy and the 2nd (17 cases) receiving placebo. Steroid receptors were evaluated in the nuclear (n) and cytosolic (c) fraction using the saturation analysis technique (Scatchard analysis or single saturating-dose assay) for androgen (AR) and estrogen (ER) receptors and the enzyme immunoassay (EIA) for ER and progesterone receptors (PgR). Scatchard analysis of ERc and ERn revealed the presence of two classes of binding sites, one with high-affinity low-capacity binding and the other with low-affinity high-capacity binding. In the untreated BPH group, ER were higher in the n than in the c fraction: ERn were positive in 14 cases and ERc in 12 of 17 cases. In the BPH group treated with S. repens extract on the contrary, ERn were negative for both binding classes in 17 cases and ERc in 6 of 18 cases. Using EIA, ERn and ERc were detected in all 15 samples examined, but in the treated group, ERn were significantly (p less than 0.01) lower than in the untreated group, whilst ERc remained almost unchanged. Similar results were obtained measuring PgR: the n fraction of the treated group prostatic samples was significantly (p less than 0.01) lower than that of the untreated group.(ABSTRACT TRUNCATED AT 250 WORDS)     

Understanding the role of estrogen in the development of benign prostatic hyperplasia

Introduction

Benign prostatic hyperplasia (BPH) is a non-malignant enlargement of the prostate that affects ageing men. As the number of men affected by this condition will only continue to grow with the aging population, finding new strategies and new therapeutic options for its treatment is crucial. Androgenic hormones have been known to play an important role in the development of BPH and they have been a target in its medical treatment. Estrogens have also been implicated in BPH but in contrast to androgens, the functions of estrogens in the prostate are still obscure.

胰腺癌中雌激素受体表达与其生物学特性的研究

探讨胰腺癌中雌激素受体（ＥＲ）的表达及与其生物学特性的关系。方法观察２４例胰腺癌切除标本的ＥＲ表达（Ｓ－Ｐ免疫组化染色法）与肿瘤大小、有无血管浸润、淋巴转移关系,并以正常胰腺组织标本作为对照。结果２４例胰腺癌病人ＥＲ阳性１４例（５８．３％）,１２例正常胰腺中仅有１例为ＥＲ阳性（８３％）。胰腺癌的ＥＲ表达明显高于正常胰腺组织（Ｐ＜０．０１）。其中肿瘤越大其阳性表达越低,＞５ｃｍ者ＥＲ阳性率明显低于≤５ｃｍ者（Ｐ＜０．０５）。高分化胰腺癌中ＥＲ阳性率明显高于低分化者,两者之比为８３．３３％：３３．３％,Ｐ＜０．０５。ＥＲ阳性者的血管侵犯率明显低于ＥＲ阴性者（Ｐ＜０．０５）。ＥＲ阳性的胰腺癌转移率低于ＥＲ阴性者,但两者差异无显著意义（Ｐ＞０．０５）。结论胰腺癌组织中ＥＲ有较高的表达率（５３．３％）。ＥＲ的表达与肿瘤的大小、细胞分化程度、血管浸润等生物学特性相关,ＥＲ阳性者多有较好的生物学特性。     

Serum hormone levels in benign prostatic hyperplasia.

The peripheral serum levels of FSH, LH, testosterone and total estrone (sum of free + conjugated estrone, mainly estrone sulphate) were determined in 20 patients with benign prostatic hyperplasia (BPH) aged 59--79 years and in 49 healthy men aged 58--79 years. There were no significant differences in the levels of FSH, LH and testosterone. The level of total estrone was significantly higher in the BPH group (p less than 0.05). The results are in accordance with our previous study on the urinary estrogen excretion in BPH and give additional support to the hypotheses concerning a role of the estrogens in the etiology of BPH.     

Detection of estrogen receptor in paraffin-embedded sections of breast carcinoma by immunohistochemistry and in situ hybridization.

Biopsy specimens of small breast carcinomas are often insufficient for both diagnosis and the biochemical determination of estrogen receptor (ER) protein. Recent reports from various laboratories have shown the utility of immunohistochemical detection of ER protein in paraffin-embedded tissue sections. We used immunohistochemical (IHC) staining with the Abbott ER antibody (H222) and in situ hybridization (ISH) analysis with a 35S-labeled and a biotinylated oligonucleotide probe to detect ER protein and messenger RNA (mRNA) in tissue sections of 53 breast carcinomas. The dextran-coated charcoal (DCC) assay of these same cases revealed positive receptor levels in 31 of 53 cases, whereas the IHC method was positive in 33 of 53 cases. ISH for detection of ER mRNA was more sensitive than IHC or the biochemical assay for estrogen binding proteins, as the isotopic probe detected ER mRNA in 47 of 53 cases, whereas the biotinylated probe detected ER mRNA in 46 of 53 cases. These results indicate that ER protein can be readily detected in enzyme-treated paraffin tissue sections and that the IHC detection of ER protein correlates highly with the DCC assay. ISH with isotopic and biotinylated probes detects ER mRNA in most cases found to be positive for ER protein and also in many cases without detectable ER protein. Although detection of ER protein in paraffin sections correlates highly with the biochemical assay in this report and in other reported studies, the clinical significance of increased sensitivity by ISH is unknown and must await clinical correlative follow-up studies.     

Cellular distribution of retinoic acid receptor-alpha in benign hyperplastic and malignant human prostates: comparison with androgen, estrogen and progesterone receptor status

OBJECTIVES: Retinoids are unique modulators of gene activity, cell growth and differentiation by binding to a series of nuclear receptors, i.e. all-trans-retinoic acid receptors (RAR) or 9-cis-retinoid receptors (RXR). In this study, the expression of RARalpha was immunohistochemically evaluated in benign, hyperplastic and malignant prostatic tissue and correlated with sex steroid receptor status. METHODS: Twenty-four cases of BPH and 139 cases of primary prostatic carcinoma were evaluated for RARalpha expression in correlation with androgen (AR), estrogen (ER) and progesterone (PGR) receptor staining, as well as with tumor grade. RESULTS: RARalpha was detected in the nuclei of epithelial cells in both BPH and prostate carcinoma cases. A modest inverse relationship with grade was present, especially for grade I and grade II tumors. AR staining was intense and a strong inverse relationship with grade was revealed. Although ER and PGR showed nuclear staining in prostatic epithelium, the overall expression for these receptors was low. When RARalpha content was compared to the nuclear AR expression, at least two-fold higher RARalpha levels were observed in AR+ grade II and grade III tumors. CONCLUSIONS: RARalpha expression can be immunohistochemically evaluated in formalin-fixed paraffin-embedded prostatic tissue. RARalpha expression is significantly elevated in AR+ moderately and poorly differentiated prostate carcinomas. Immunohistochemical determination of RARalpha content may be useful in defining the patient subsets in which retinoid-based treatment may be of clinical value.     

Upregulation of estrogen and androgen receptors modulate expression of FGF-2 and FGF-7 in human,cultured, prostatic stromal cells exposed to high concentrations of estradiol

This study was performed to develop an experimental model in which expression of estrogen receptors (ER) by human prostatic stromal cells could be reproducibly enhanced relative to similar cells with low ER expression. The second aim was to characterise changes in expression of ER, androgen receptor (AR), FGF-2 and FGF-7 in stromal cells exposed to high and low concentrations of estradiol and testosterone mimicking the different sex hormone levels between young and elderly men. Five strains of human prostatic stromal cells, isolated from BPH resections, were grown in steroid-free medium plus 1 micromol 17beta-estradiol. After 10 days, cells were passaged and grown in the same medium without estradiol until confluent. In a second study four cell strains were exposed to high and low concentrations of 17beta-estradiol and testosterone for 10 days. Cells were labelled with fluorescent antibodies to ERalpha, AR, FGF-2 and FGF-7 and the fluorescence intensity measured by flow cytometry. Following exposure to 1 micromol estradiol, stromal cells showed reduced expression of AR and ERalpha but after passage without estradiol they showed a 25% increase in both receptors over controls. Different combinations of sex hormones induced inconsistent changes with respect to expression of ER, AR and FGFs in the various cell-strains. However, there was a highly significant correlation between AR, ER and FGF-2 and FGF-7, which was cell strain-specific. Thus, changes in sex hormone balance per se may not be solely responsible for the observed increases in prostatic ER levels in BPH. Since expression of ER is correlated with synthesis of FGF-2 and FGF-7, it is likely that increases in stromal ER may mediate the synthesis of stromally-derived growth factors which contribute to the aetiopathogenesis of benign prostatic hyperplasia.