Androgen Receptor Staining Research Articles

Expression of Aromatase, Estrogen Receptor α and β, Androgen Receptor, and Cytochrome P-450scc in the Human Early Prepubertal Testis

The expression of aromatase, estrogen receptor alpha (ERalpha) and beta (ERbeta), androgen receptor (AR), and cytochrome P-450 side chain cleavage enzyme (cP450scc) was studied in prepubertal testis. Samples were divided in three age groups (GRs): GR1, newborns (1- to 21-d-old neonates, n = 5); GR2, postnatal activation stage (1- to 7-mo-old infants, n = 6); GR3, childhood (12- to 60-mo-old boys, n = 4). Absent or very poor detection of ERalpha by immunohistochemistry in all cells and by mRNA expression was observed. Leydig cells (LCs) of GR1 and GR2 showed strong immunostaining of aromatase and cP450scc but weak staining of ERbeta and AR. Interstitial cells (ICs) and Sertoli cells (SCs) expressed ERbeta, particularly in GR1 and GR2. Strong expression of AR was found in peritubular cells (PCs). For all markers, expression in GR3 was the weakest. In germ cells (GCs), i.e. gonocytes and spermatogonia, aromatase and ERbeta were immunoexpressed strongly whereas no expression of ERalpha, AR, or cP450scc was detected. It is proposed that in newborn and infantile testis, testosterone acting on PCs might modulate infant LC differentiation, whereas the absence of AR in SCs prevents development of spermatogenesis. The role of estrogen is less clear, but it could modulate the preservation of an adequate pool of precursor LCs and GCs.

P16, p14, p53, cyclin D1, and steroid hormone receptor expression and human papillomaviruses analysis in primary squamous cell carcinoma of the endometrium

Pathogenetically, endometrioid adenocarcinomas of the endometrium are associated with hyperestrogenism and serous papillary carcinomas with alterations of p53. The etiology of primary endometrial squamous cell carcinoma (ESCC), however, is speculative. The purpose of this study was to evaluate the role of p14, p16, p53, cyclin D1, steroid hormone receptors, and human papillomaviruses (HPV) infection in the pathogenesis of primary endometrial squamous cell carcinoma. The expression of p16, p14, p53, cyclin D1, and steroid hormone receptors (estrogen, progesterone, and androgen) was examined immunohistochemically in 8 primary ESCCs. HPV analysis was performed using general primers and HPV typing. The median age of the patients was 62.1 years. Four cases showed positive nuclear and cytoplasmic p16 staining in an insular pattern, and 1 case nuclear positivity for p53 and estrogen receptors, respectively. Four of 8 cases were positive for progesterone receptor analysis and cyclin D1. All cases were negative for p14 and androgen receptor staining. All but one case were negative for HPV analysis. Five patients were alive with and without evidence of disease after a mean follow-up of 6.1 years. The results of this study suggest that alterations of the p16 pathway may play an etiologic role in at least a proportion of the ESCC, but without any association to HPV infection. Factors known to play a pathogenetic role in types 1 and 2 of endometrial carcinomas are not associated with primary ESCC. However, prognostically, ESCCs are more related to type 1 cancers.

Intensity of Androgen and Epidermal Growth Factor Receptor Immunoreactivity in Samples of Radical Prostatectomy as Prognostic Indicator: Correlation With Clinical Data of Long-Term Observations

We analyzed the potential prognostic significance of the immunohistochemical expression of androgen and growth factor receptors determined in prostatectomy specimens of patients with prostate cancer. A cohort of 211 patients with locally confined prostate cancer treated with radical prostatectomy with or without antiandrogen pretreatment between January 1, 1990 and August 31, 1996 was observed prospectively. Prostatectomy samples were processed immunohistochemically to visualize androgen and growth factor receptors, of which immunoreaction intensity was scored relative to that of positive control tissue. Clinical postoperative data were processed using the Kaplan-Meier method, log rank test, and univariate and multivariate explorative Cox modeling to evaluate the contribution to overall and relapse-free survival. There were statistical dependencies between the androgen receptor and epidermal growth factor receptor staining indexes. Following data stratification according to the epidermal growth factor receptor staining index the prognosis associated with a low androgen receptor staining index was worse than that with a higher androgen receptor staining index. Cox regression analysis for relapse-free survival confirmed that the risk factors low androgen receptor and increased epidermal growth factor receptor staining were associated with significantly increased relative risk. Univariate Kaplan-Meier analysis showed that patients with grade 3 carcinoma had a worse prognosis than those with better differentiated carcinoma, whereas antiandrogen pretreatment had no influence on overall survival or relapse-free survival. Using a multivariate proportional hazards regression model for data on a cohort of 211 patients with 68 showing relapse/progress or death from disease a low intensity of androgen receptor staining indicated a poor prognosis.

Analysis of hormonal receptor status in tissue from prostate cancer that developed 26 years after bilateral orchiectomy and following 41 years of estrogen therapy

14610 Background: The development of prostate cancer in the absence of normal male testosterone levels and in the presence of elevated estrogen levels is rare and poorly understood. Methods: The androgen, estrogen and progesterone receptors were evaluated in prostate cancer tissue from a 60-year-old, long-term transgender woman. Her serum hormone levels were assessed and the prostate specific antigen (PSA) level was followed during hormonal and radiation treatment. This patient began estrogen therapy at age 19 and underwent bilateral orchiectomy at age 34 during gender re-assignment surgery. Work-up for gross hematuria revealed a Gleason 8, locally advanced prostate cancer. Results: The tumor showed abundant staining for androgen receptors (AR), with a nuclear pattern consistent with hormone refractory prostate cancer. The tumor did not stain for estrogen receptors (ER) or progesterone receptors (PR). Compared to reference values for men, this transgender woman had low serum testosterone (44 ng/dl) and free testosterone (0.9 pg/mL) levels and elevated estradiol (53 pg/mL) and estrone (377 pg/mL) levels. Prior to the initiation of radiation, treatment with bicalutamide and dutasteride reduced her PSA from 240 ng/ml to 77 ng/ml at week 6 (see Table). After treatment with external beam radiation her PSA fell to 4.6 ng/mL. At week 24, her PSA was 0.8 ng/mL. Conclusions: This high-grade, locally advanced prostate cancer in a transgender woman represents the longest reported duration of a castrate state preceding the development of prostate cancer, exceeding prior reports by several decades. The AR positive and ER/PR negative status of this tumor in the setting of low testosterone and elevated estrogen levels challenge long-standing paradigms regarding the testosterone requirement for prostatic carcinogenesis. In addition, these findings raise questions about the role of estrogens in prostate cancer. [Table: see text] [Table: see text]

Energy balance alters dunning R3327-H prostate tumor architecture, androgen receptor expression, and nuclear morphometry in rats

We hypothesize that diet or energy restriction may alter prostate tumor architecture and nuclear morphometry in a manner predictive of indolent biology. Adult male rats bearing the Dunning R3327-H tumor were provided free-access to the AIN76 diet or 30% diet/energy restriction for 16 weeks. Tumor tissue was harvested and stained for biomarker evaluation by quantitative digital image analysis. Compared to controls, tumors from restricted rats showed a contracted stromal compartment with an increased epithelial/glandular area, and a greater number of smaller and more homogeneous glands. In parallel, the nuclei in tumors harvested from diet-restricted rats were smaller in size, showed a lower proportion with androgen receptor (AR) staining, and reduced intensity of AR staining. Our observations support the hypothesis that diet/energy restriction alters prostate tumor architecture, AR expression, and nuclear morphology consistent with a more differentiated and less aggressive phenotype.

Large Cell Neuroendocrine Carcinoma of Prostate

Neuroendocrine (NE) differentiation in prostate cancer is typically detected by immunohistochemistry as single cells in conventional adenocarcinoma. Prostatic NE tumors, such as carcinoid or small cell carcinoma, are rare and large cell NE carcinoma (LCNEC) is described only in case reports. We identified 7 cases of LCNEC and compiled their clinicopathologic characteristics. In 6 cases, there was a history of adenocarcinoma treated with hormone therapy for a mean of 2.4 years (range: 2 to 3 y). The remaining case was de novo LCNEC. LCNEC was incidentally diagnosed in palliative transurethral resection specimens in 5 cases. The mean patient age at diagnosis with LCNEC was 67 years (range: 43 to 81 y). LCNEC comprised solid sheets and ribbons of cells with abundant pale to amphophilic cytoplasm, large nuclei with coarse chromatin and prominent nucleoli along with brisk mitotic activity and foci of necrosis. In 6 cases, there were foci of admixed adenocarcinoma, 4 of which showed hormone therapy effects. LCNEC was strongly positive for CD56, CD57, chromogranin A, synaptophysin, and P504S/alpha methylacyl CoA racemase. There was strong bcl-2 overexpression, expression of MIB1, and p53 in >50% of nuclei, focally positive staining for prostate specific antigen and prostatic acid phosphatase and negative androgen receptor staining. Follow-up was available for 6 patients, all of who died with metastatic disease at mean of 7 months (range: 3 to 12 mo) after platinum-based chemotherapy. LCNEC of prostate is a distinct clinicopathologic entity that typically manifests after long-term hormonal therapy for prostatic adenocarcinoma and likely arises through clonal progression under the selection pressure of therapy.

Effect of dexamethasone on serum and testicular concentrations of testosterone in stallions

Distribution of androgen receptors (ARs) in the epididymal duct and prostate of three entire stallions and one bilaterally cryptorchid horse was studied immunohistochemically using a polyclonal rabbit antiserum against the ARs. In both the healthy stallions and the cryptorchid, the epithelial cells of the epididymides showed nuclear staining for ARs. The intensity of AR-staining in the principal cells of the epididymis was stronger than that of the basal cells. In the prostate, the glandular secretory cells were moderately stained whereas the basal cells expressed weak AR-staining. Immunostaining for ARs in the reproductive tissues of the cryptorchid horse was always stronger than in those of the stallions. Our results demonstrate for the first time the AR localisation to equine epididymal and prostatic cells, which are directly regulated by androgens.

Cell-specific Regulation of Androgen Receptor Phosphorylation in Vivo

The biological ramifications of phosphorylation of the androgen receptor (AR) are largely unknown. To examine the phosphorylation of AR at serine 213, a putative substrate for Akt, a phosphorylation site-specific antibody was generated. The use of this antibody indicated that AR Ser-213 is phosphorylated in vivo and that phosphorylation is tightly regulated in a cell type-specific manner. Furthermore, Ser-213 phosphorylation took place with rapid kinetics and was inhibited by the phosphatidylinositol 3-kinase inhibitor LY294002. Phosphorylation occurred in response to R1881 and dihydrotestosterone but weakly if at all in response to testosterone. It did not occur in response to AR antagonists or growth factor stimulation in the absence of an AR agonist. Transcription assays using an AR-responsive reporter gene construct showed that activated phosphatidylinositol 3-kinase inhibited transcription mediated by wild type AR but not that of a mutant AR variant (S213A), which could not be phosphorylated at Ser-213. By immunohistochemistry, the AR Ser(P)-213 antigen was detected in prostate epithelial but not stromal cells despite the fact that an antibody recognizing both phosphorylated and non-phosphorylated forms of AR demonstrates that AR is present in both cell types as expected. In fetal tissue the AR-Ser(P)-213 antigen was present in epithelial cells of the urogenital sinus when endogenous androgen levels were high and activated Akt was prevalent, but absent at a later stage of development when endogenous androgen levels were low and Akt activation was minimal. Immunoreactivity was evident in differentiated cells lining the lumen of the urogenital sinus but not in rapidly dividing, Ki67 positive cells within the developing prostate or stromal tissue, suggesting that site-specific phosphorylation of AR Ser-213 by cellular kinases occurs in a non-proliferating cellular milieu.

Neurodistribution of Androgen Receptor Immunoreactivity in the Male Frog, Rana esculenta

Sexual behavior in vertebrates depends on the cyclic release of steroids and their binding to the brain receptors. Previously, we demonstrated the presence of specific binding of (3)H-testosterone and staining with PG-21 in the brain of the adult male frog, Rana esculenta. Here, we report our further receptor characterization using an anti-androgen receptor antiserum, PG-21, and the androgen site of action in frog brain. Nuclei, which contained cells labeled for the androgen receptor (AR), were mainly identified in the olfactory bulbs, preoptic-septal region, infundibulum, amygdala, thalamus, tectum, torus semicircularis, and medulla. The neuroanatomical AR staining appears similar to that in other lower vertebrates.

The presence of androgen receptors in the epididymis and prostate of the stallion and cryptorchid horse – A preliminary study

Distribution of androgen receptors (ARs) in the epididymal duct and prostate of three entire stallions and one bilaterally cryptorchid horse was studied immunohistochemically using a polyclonal rabbit antiserum against the ARs. In both the healthy stallions and the cryptorchid, the epithelial cells of the epididymides showed nuclear staining for ARs. The intensity of AR-staining in the principal cells of the epididymis was stronger than that of the basal cells. In the prostate, the glandular secretory cells were moderately stained whereas the basal cells expressed weak AR-staining. Immunostaining for ARs in the reproductive tissues of the cryptorchid horse was always stronger than in those of the stallions. Our results demonstrate for the first time the AR localisation to equine epididymal and prostatic cells, which are directly regulated by androgens.

Increased phospho-AKT is associated with loss of the androgen receptor during the progression ofN-methyl-N-nitrosourea-induced prostate carcinogenesis in rats

Characterization of molecular events during N-methyl-N-nitrosourea (MNU)-induced rat prostate carcinogenesis enhances the utility of this model for the preclinical assessment of preventive strategies. Androgen independence is typical of advanced human prostate cancer and may occur through multiple mechanisms including the loss of androgen receptor (AR) expression and the activation of alternative signaling pathways. We examined the interrelationships between AR and p-AKT expression by immunohistochemical staining during MNU-androgen-induced prostate carcinogenesis in male Wistar-Unilever rats. Histone nuclear staining and image analysis was employed to assess parallel changes in chromatin and nuclear structure. The percentage of AR positive nuclei decreased (P < 0.01) as carcinogenesis progressed: hyperplasia (92%), atypical hyperplasia (92%), well-differentiated adenocarcinoma (57%), moderately-differentiated adenocarcinoma (19%), and poorly-differentiated adenocarcinoma (10%). Conversely, p-AKT staining increased significantly during carcinogenesis. Sparse staining was observed in normal tissues (0.2% of epithelial area) and hyperplastic lesions (0.1%), while expression increased significantly (P < 0.001) in atypical hyperplasia (7.6%), well-differentiated adenocarcinoma (16.7%), moderately-differentiated adenocarcinoma (19.6%), and poorly-differentiated adenocarcinoma (17.4%). In parallel, nuclear morphometry revealed increased nuclear size, greater irregularity, and lower DNA compactness as cancers became more poorly differentiated. In the MNU model, the progressive evolution of dominant tumor cell populations showing an increase in p-AKT in parallel with a decline in AR staining suggests that activation of AKT signaling may be one of several mechanisms contributing to androgen insensitivity during prostate cancer progression. Our observations mimic findings suggested by human studies and support the relevance of the MNU model in preclinical studies of preventive strategies.

Cystic hypersecretory carcinoma: rare and poorly recognized variant of intraductal carcinoma of the breast. Report of five cases

To report five cases of a rare variant of intraductal carcinoma of the breast, so-called cystic hypersecretory carcinoma. The clinical and pathological characteristics of the lesion are described, along with a review of the literature. The patients were females aged between 53 and 78 years (average 66.8 years). The size of the lesions ranged between 70 and 80 mm in largest dimension. In two cases, the development of high-grade invasive ductal carcinoma was observed; in one additional case there was recurrence of high-grade in-situ carcinoma after 3 years. This emphasizes the importance of correct diagnosis of this potentially aggressive lesion. Strong over-expression of HER-2/neu protein was observed in three cases, including the two with an invasive component. Protein p53 was variably positive in all cases. Steroid receptor immunohistochemistry yielded variable results with only one case being positive for both oestrogen and progesterone receptors. Interestingly, in most cases (4/5) staining for androgen receptors was observed. Cystic hypersecretory ductal carcinoma of the breast is a rare distinctive variant of ductal carcinoma in situ. It has the potential for invasive growth and the development of metastases.

Immunohistochemical localization of androgen receptors in the urogenital tracts of human embryos.

The aim of this study was to investigate androgen receptor (AR) expression in the developing human urogenital tract. The distribution of AR was examined in paraffin-embedded tissue sections of the lower urogenital tract using 55 human embryos of 8-12 weeks of gestation. Immunohistochemistry was performed for AR detection and gender was determined by polymerized chain reaction. There were no differences in the distribution of AR in male and female embryos at any stage of gestation. AR was present only in the mesenchymal tissues of the urogenital sinus at 8 weeks whilst the epithelium was negative, but after 9 weeks the epithelium also showed progressively more positive staining. In the phallus, AR staining was prominent. There was far less staining in the epithelium of the urethral groove from 8 to 10 weeks, whilst the mesenchyme of the urethral folds showed positive staining. At 11 and 12 weeks, both the urethral groove and folds showed uniform staining. The genital tubercle, genital swelling and bulbourethral gland precusors were also positively stained, although paramesonephric ducts were negative. Staining was observed in the mesonephric duct from 9 weeks. There was an absence of staining in the rectum at all stages of gestation. The expression of AR in an epithelium may be dependent upon the mesenchyme. Mesenchymal-epithelial interactions played an important role in development, as has been described in experimental animals. AR expression could play a part in the growth of the genital organs.

Androgen receptor expression is inversely correlated with pathologic tumor stage in bladder cancer

Objectives To evaluate the expression of the androgen receptor (AR) in transitional cell carcinoma (TCC) of the bladder, and to assess whether its expression correlated with pathologic tumor stage. TCC of the bladder is three times more common in males than in females. The origin of this sex difference in incidence is unknown. Methods We evaluated tumor specimens from 49 consecutive patients treated for TCC of the bladder at our institution between July 2002 and June 2003. Immunohistochemistry was performed using a monoclonal mouse anti-AR antibody on paraffin-embedded tissue sections of tumors obtained from transurethral resection, radical cystectomy, or resection of metastases. Specimens were assessed for AR expression, and, in tumors that demonstrated AR staining, the percentage of nuclei that stained positive was recorded. Results Of the 49 tumors, 26 (53.1%) expressed the AR. The percentage of tumors that expressed the AR decreased with increasing pathologic stage, from 88.9% of pTa lesions to 0% of pT3 tumors. Overall, 75% of superficial tumors (pTa + pT1 + carcinoma in situ) expressed the AR compared with 21.4% of invasive tumors (pT2 + pT3; P = 0.002). In addition, among AR-expressing tumors, the mean percentage of nuclei that stained positive for the AR was significantly greater in pTa tumors (62.5%) than in pT1 (31%) or pT2 (20%) tumors ( P = 0.005). Conclusions We found a decrease in AR protein expression in tumors with increased pathologic stage. Our data suggest that the loss of AR expression is associated with invasive bladder cancer.

(Anti-) androgenic reactions in the rat prostate and seminal vesicle: Validation of proliferation markers and androgen receptor staining pattern and the effects of the „xenoestrogen“ BPA in the Wistar rat.

Androgenic effects of xenoestrogens are now in the focus of research: Therefore, the aim of this study was to validate proliferating activities using immunohistochemistry and morphometry for the evaluation of androgen-like effects of the xenoestrogen bisphenol A. Bisphenol A is an industrial monomer used in the production of polycarbonate plastics and epoxy resins. Orchiectomized Wistar rats (the negative control group), (n=13) were treated p.o. with 3, 50, 200, 500mg/kg bw/day for seven days. The positive control group was treated s.c. with 1mg/kg bw/day testosterone propionate (TP). Flutamide (3mg/kg bw/day p.o.) was used to antagonize possible androgen effects of the „high dose“ of BPA. Immunohistochemical staining and proliferating activity were determined morphometrically using the Axiophot of Zeiss-Vision (KS 300, KS 100, KS RUN). For the densitometric evaluation of the androgen receptor staining we used a polyclonal antibody (sc-815, Santa Cruz) as well as a monoclonal antibody (No. 554224, BD PharMingen). For the evaluation of the proliferation we used two different primary antibodies from DakoCytomation (Ki-67 antigen, Clone MIB-5) and from Novocastra (PCNA, Clone PC10). Counting the number of immunoreactive cells (N=1000 per [13] sections) in prostate the following data were achieved: 1. PCNA: Intact 90%, Orchiectomized 10%, TP substituted orchiectomized rats 88%, 3mg/kg bw BPA 3%, 50mg/kg bw BPA 4%, 200mg/kg bw BPA 3%, 500mg/kg bw BPA 5%, 500+FL BPA 5% immunoreactive cells. 2. MIB-5: Intact 85%, Orchiectomized 8%, TP substituted orchiectomized rats 90%, 3mg/kg bw BPA 11%, 50mg/kg bw BPA 9%, 200mg/kg bw BPA 3%, 500mg/kg bw BPA 2%, 500+FL BPA 1% immunoreactive cells. In view of the validation of the proliferation markers analyzed and the reaction of the androgen receptor it can be concluded that the proliferation markers proofed to indicate androgenic reactions stimulating proliferation activity more sensitive. BPA showed in all doses tested no stimulation of the proliferating activity on prostate cells compared with the castrated control.

Androgen receptor expression in relation to apoptosis and the expression of cell cycle related proteins in prostate cancer.

The expression of several genes involved in the regulation of cell cycle and apoptosis may be regulated via the androgen receptor (AR) in the prostate. AR may have a role in the prognosis of prostatic carcinoma. The aim was to examine AR expression status and its relationship with markers of proliferation, apoptosis and cell cycle control in prostate cancer. Expression of AR, bcl-2, bax, Ki-67 and p53 was examined in paraffin-embedded tissues from 50 cases of prostate carcinoma by immunohistochemistry and evaluated using an index of staining. Detection of apoptotic cells was performed by TUNEL method. Correlation between AR expression and apoptosis, proliferation index, bcl-2, bax and p53 and also clinicopathological parameters including stage, pathological grade and Gleason score were determined. AR expression was observed in all cases with mean expression of 81%+/-15 and mean staining index of 141+/-65. No correlation was found between AR expression and apoptosis detected in patients. The mean AR staining index was 170+/-72 in bcl-2 positive tumors versus 120+/-53 in bcl-2 negative tumors showing a significant association between AR and bcl-2 expression (p=0.015). AR expression also showed a significant association with bcl-2/bax ratio (r=0.321, p=0.023) and Ki-67 proliferation staining index (r=0.396, p=0.004). Although a significant correlation between Ki-67 and p53 with differentiation status of the tumors was observed (p<0.004) no correlation was found with AR. AR expression showed no prognostic value regarding its correlation with stage and differentiation status of the prostate carcinoma. However, its significant correlation with Ki-67 and bcl-2 that are markers of cell survival suggest its contribution to tumor cell progression.

Computer based receptogram approach: an objective way of assessing immunohistochemistry of androgen receptor staining and its correlation with hormonal response in metastatic carcinoma of prostate

Aims: To categorise the immunostaining heterogeneity of androgen receptors in metastatic carcinoma of the prostate using a pattern oriented approach and to correlate the results with response to hormonal treatment....

Characterization of a transplantable hormone-responsive human prostatic cancer xenograft TEN12 and its androgen-resistant sublines.

Models for human prostate cancer can facilitate the study of resistance to endocrine therapy, aid drug discovery, and pre-clinical assessment. Characteristics thought relevant to the growth in athymic nude mice of TEN12, an androgen-dependent transplantable prostatic cell line derived from a primary prostate carcinoma, and its two androgen-independent sublines, TEN12F and TEN12C, have been assessed immunocytochemically. The xenografts of the parental TEN12 line are moderately differentiated with both papillary and glandular regions, pleomorphic nuclei and abundant mitotic figures and are extremely vascular. The cells express androgen receptor (AR), PSA, VEGF, EGFR, c-erbB2, and TGFalpha. Both TEN12F and TEN12C xenografts possessed a more anaplastic morphology and displayed significantly lower growth rates, reduced blood vessel density (BVD), decreased MIB-1 antigen and E-cadherin expression and increased cytoplasmic AR and HSP90 staining. Elevated EGFR (membrane) but not c-erbB2 expression was demonstrated in the TEN12F line only. Castration of mice bearing TEN12 xenografts rapidly induced the appearance of cytoplasmic AR in the cells, PSA levels decreased initially but recovered to below pre-castration levels whilst reduced TGFalpha and loss of VEGF expression was seen in the long-term castrates. TEN12 and its sublines offer additional in vivo models to study the factors involved in the progression of prostatic cancer to androgen-independence.

Estrogen receptor expression in meibomian glands and its correlation with age and dry-eye parameters

To investigate the correlation between estrogen receptor-positive basal cells of the meibomian glands of the lower lid with age, gender, and subjective and objective dry-eye parameters. Sixteen lower lid specimens were collected from 7 female and 9 male patients (age range 22-88 years) during tumor surgery requiring whole-thickness excision. Prior to surgery the patients were asked about any dry-eye symptoms (score 0-4, where 0=no dry-eye symptoms). Tear break-up time was measured, and Schirmer I and II tests were performed. We obtained histological sections from the outer margins of the formalin-fixed and paraffin-embedded specimens and performed immunohistochemical staining for estrogen, progesterone and androgen receptors. At least 500 cells were counted per specimen, and the proportion of positively stained cells was calculated and correlated to the age and dry-eye parameters. All meibomian glands had positive nuclear staining with antibodies for estrogen receptors in their outer cell layers, i.e., the basal cell layer. The proportion of cells expressing estrogen receptors increased with age independent of gender (correlation coefficient=0.67, P<0.005). No correlation was found between estrogen receptor positivity and subjective dry-eye symptoms, tear break-up time, or Schirmer I and II test results. There was no difference in the proportion of cells expressing estrogen receptors between female and male patients (P=0.5). The amount of meibomian gland cells expressing estrogen receptors in the lower lid seems to increase with age independent of gender and seems not to affect the fat layer and stability of the tear film.

Immunocytochemical Localization of Glutathione-Peroxidase (GSH-PO) and Bcl-2 in the Rat Ventral Prostate

In order to confirm the relationship between glutathione-peroxidase (GSH-PO) and Bcl-2, we have studied the immunocytochemical localization of GSH-PO, Bcl-2, androgen receptor (AR) and apoptosis in the rat ventral prostatic cells in the presence and absence of testosterone. Male Crj:CD(SD) IGS rats were divided into four experimental groups. Group 1 consisted of untreated controls. In group 2, rats were sacrificed two days after castration. In groups 3 and 4, rats were subcutaneously administered 1 mg/animal of testosterone daily for 3 or 7 days after two days of castration, respectively. In group 1, GSH-PO was predominantly observed in the glandular epithelial cells. Bcl-2 was localized exclusively in the cytoplasm of the glandular epithelial cells. AR was localized in the nuclei of the glandular epithelial cells. In group 2, the intensity of the staining of GSH-PO, Bcl-2 and AR markedly decreased. However, the number of apoptotic cells (apoptotic index) markedly increased. In groups 3 and 4, the immunoreactivities of GSH-PO, Bcl-2 and AR clearly recovered. These findings strongly suggest that the expression of GSH-PO and Bcl-2 in the glandular epithelial cells of the rat ventral prostate is testosterone-dependent. Furthermore, co-expression of GSH-PO and Bcl-2 in the prostatic cells are considered to be normal or adaptive aspects of the cells.