Human Benign Prostatic Hyperplasia Research Articles

PDE5 inhibitors blunt inflammation in human BPH: A potential mechanism of action for PDE5 inhibitors in LUTS

Metabolic syndrome (MetS) and benign prostate hyperplasia (BPH)/low urinary tract symptoms (LUTS) are often comorbid. Chronic inflammation is one of the putative links between these diseases. Phosphodiesterase type 5 inhibitors (PDE5i) are recognized as an effective treatment of BPH-related LUTS. One proposed mechanism of action of PDE5 is the inhibition of intraprostatic inflammation. In this study we investigate whether PDE5i could blunt inflammation in the human prostate. Evaluation of the effect of tadalafil and vardenafil on secretion of interleukin 8 (IL-8, a surrogate marker of prostate inflammation) by human myofibroblast prostatic cells (hBPH) exposed to different inflammatory stimuli. We preliminary evaluate histological features of prostatic inflammatory infiltrates in BPH patients enrolled in a randomized, double bind, placebo controlled study aimed at investigating the efficacy of vardenafil (10 mg/day, for 12 weeks) on BPH/LUTS. In vitro treatment with tadalafil or vardenafil on hBPH reduced IL-8 secretion induced by either TNFα or metabolic factors, including oxidized low-density lipoprotein, oxLDL, to the same extent as a PDE5-insensitive PKG agonist Sp-8-Br-PET-cGMP. These effects were reverted by the PKG inhibitor KT5823, suggesting a cGMP/PKG-dependency. Treatment with tadalafil or vardenafil significantly suppressed oxLDL receptor (LOX-1) expression. Histological evaluation of anti-CD45 staining (CD45 score) in prostatectomy specimens of BPH patients showed a positive association with MetS severity. Reduced HDL-cholesterol and elevated triglycerides were the only MetS factors significantly associated with CD45 score. In the MetS cohort there was a significant lower CD45 score in the vardenafil-arm versus the placebo-one.

Applicators for Magnetic Resonance–Guided Ultrasonic Ablation of Benign Prostatic Hyperplasia

The aims of this study were to evaluate in a canine model applicators designed for ablation of human benign prostatic hyperplasia (BPH) in vivo under magnetic resonance imaging (MRI) guidance, including magnetic resonance thermal imaging (MRTI), determine the ability of MRI techniques to visualize ablative changes in prostate, and evaluate the acute and longer term histologic appearances of prostate tissue ablated during these studies. An MRI-compatible transurethral device incorporating a tubular transducer array with dual 120° sectors was used to ablate canine prostate tissue in vivo, in zones similar to regions of human BPH (enlarged transition zones). Magnetic resonance thermal imaging was used for monitoring of ablation in a 3-T environment, and postablation MRIs were performed to determine the visibility of ablated regions. Three canine prostates were ablated in acute studies, and 2 animals were rescanned before killing at 31 days postablation. Acute and chronic appearances of ablated prostate tissue were evaluated histologically and were correlated with the MRTI and postablation MRI scans. It was possible to ablate regions similar in size to enlarged transition zone in human BPH in 6 to 18 minutes. Regions of acute ablation showed a central "heat-fixed" region surrounded by a region of more obvious necrosis with complete disruption of tissue architecture. After 31 days, ablated regions demonstrated complete apparent resorption of ablated tissue with formation of cystic regions containing fluid. The inherent cooling of the urethra using the technique resulted in complete urethral preservation in all cases. Prostatic ablation of zones of size and shape corresponding to human BPH is possible using appropriate transurethral applicators using MRTI, and ablated tissue may be depicted clearly in contrast-enhanced magnetic resonance images. The ability accurately to monitor prostate tissue heating, the apparent resorption of ablated regions over 1 month, and the inherent urethral preservation suggest that the magnetic resonance-guided techniques described are highly promising for the in vivo ablation of symptomatic human BPH.

Abstract 4074: Transcription factors involved in the genesis and progression of cancer differently modulated by transforming growth factor-beta3 (TGF-Beta3) in prostate cell lines.

Abstract Transforming growth factor-beta3 (TGF-β3) is a member of a family of multifunctional cytokines that control different cellular processes including cell proliferation, morphogenesis, migration, extracellular matrix production, cytokine secretion, and apoptosis, as well as in normal physiological and disease processes. Among the various isoforms of TGF-β, TGF-β1 has been the most studied form, especially in the context of tumor development. In contrast, only few studies have been focused on the role of TGF-β3 in tumorogenesis. Recently, it has been shown that the transcription factors Yin Yang 1 (YY1) is involved in the TGF-β3 pathway. YY1 is known to be associated with cancer progression. One the potential mechanisms of this association is linked with the inactivation of p53. On this ground, the aims of this study were i) to explore the correlated expression of p53 and YY1 in a human benign prostate hyperplasia cell line (BPH-1) and in two prostate cancer cell lines, LNCaP (androgen-sensitive) and DU-145 (androgen-refractory) after treatment with different dose of TGF-β3; ii) to determine, in TGF-β3-treated prostate cell lines, the expression of a fraction of intracellular signalling pathways involved in both cell cycle progression and in apoptosis. TGF-β3 stimulation in prostate cancer cell lines showed different effects, such as YYI downregulation and p53 overexpression. These effects are also associated with a different modulation of intracellular signaling pathways (PI3K, AKT, pAKT, PTEN, Bcl-2, Bax, and iNOS). In conclusion, these findings may have important therapeutic implications in prostate cancer. Citation Format: Silvia Caggia, Saverio Candido, Massimo Libra, Venera Cardile. Transcription factors involved in the genesis and progression of cancer differently modulated by transforming growth factor-beta3 (TGF-Beta3) in prostate cell lines. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4074. doi:10.1158/1538-7445.AM2013-4074

Sex steroid receptor expression and localization in benign prostatic hyperplasia varies with tissue compartment

Androgens and estrogens, acting via their respective receptors, are important in benign prostatic hyperplasia (BPH). The goals of this study were to quantitatively characterize the tissue distribution and staining intensity of androgen receptor (AR) and estrogen receptor-alpha (ERα), and assess cells expressing both AR and ERα, in human BPH compared to normal prostate. A tissue microarray composed of normal prostate and BPH tissue was used and multiplexed immunohistochemistry was performed to detect AR and ERα. We used a multispectral imaging platform for automated scanning, tissue and cell segmentation and marker quantification. BPH specimens had an increased number of epithelial and stromal cells and increased percentage of epithelium. In both stroma and epithelium, the mean nuclear area was decreased in BPH relative to normal prostate. AR expression and staining intensity in epithelial and stromal cells was significantly increased in BPH compared to normal prostate. ERα expression was increased in BPH epithelium. However, stromal ERα expression and staining intensity was decreased in BPH compared to normal prostate. Double positive (AR and ERα) epithelial cells were more prevalent in BPH, and fewer double negative (AR and ERα) stromal and epithelial negative cells were observed in BPH. These data underscore the importance of tissue layer localization and expression of steroid hormone receptors in the prostate. Understanding the tissue-specific hormone action of androgens and estrogens will lead to a better understanding of mechanisms of pathogenesis in the prostate and may lead to better treatment for BPH.

Targeting Androgen Receptor to Suppress Macrophage-induced EMT and Benign Prostatic Hyperplasia (BPH) Development

Early studies suggested macrophages might play roles in inflammation-associated benign prostatic hyperplasia (BPH) development, yet the underlying mechanisms remain unclear. Here we first showed that CD68(+) macrophages were identified in both epithelium and the stromal area of human BPH tissues. We then established an in vitro co-culture model with prostate epithelial and macrophage cell lines to study the potential impacts of infiltrating macrophages in the BPH development and found that co-culturing prostate epithelial cells with macrophages promoted migration of macrophages. In a three-dimensional culture system, the sphere diameter of BPH-1 prostate cells was significantly increased during coculture with THP-1 macrophage cells. Mechanism dissection suggested that expression levels of epithelial-mesenchymal transition (EMT) markers, such as N-cadherin, Snail, and TGF-β2, were increased, and administration of anti-TGF-β2 neutralizing antibody during co-culture suppressed the EMT and THP-1-mediated growth of BPH-1 cells, suggesting THP-1 might go through EMT to influence the BPH development and progression. Importantly, we found that modulation of androgen receptor (AR) in BPH-1 and mPrE cells significantly increased THP-1 and RAW264.7 cell migration, respectively, and enhanced expression levels of EMT markers, suggesting that AR in prostate epithelial cells might play a role in promoting macrophage-mediated EMT in prostate epithelial cells. Silencing AR function via an AR degradation enhancer, ASC-J9, decreased the macrophage migration to BPH-1 cells and suppressed EMT marker expression. Together, these results provide the first evidence to demonstrate that prostate epithelial AR function is important for macrophage-mediated EMT and proliferation of prostate epithelial cells, which represents a previously unrecognized role of AR in the cross-talk between macrophages and prostate epithelial cells. These results may provide new insights for a new therapeutic approach to battle BPH via targeting AR and AR-mediated inflammatory signaling pathways.

Increased Infiltrated Macrophages in Benign Prostatic Hyperplasia (BPH): ROLE OF STROMAL ANDROGEN RECEPTOR IN MACROPHAGE-INDUCED PROSTATE STROMAL CELL PROLIFERATION

Infiltrated macrophages may play important roles in the development and progression of benign prostatic hyperplasia (BPH), but the underlying mechanisms remain largely unknown. We found increased macrophages infiltration in human and mouse BPH tissues. By establishing a co-culture transwell system, we found increased migration of macrophages and proliferation of prostate stromal cells during co-culture. Importantly, stromal androgen receptor (AR) could enhance the migration of macrophages and macrophage-mediated stromal cell proliferation. We identified CCL3 as an AR downstream player, and found CCL3 levels were notably increased in human and mouse BPH prostates. Ablation of prostate stromal AR in a mouse BPH model significantly reduced CCL3 expression levels in prostates. Consistently, targeting AR via an AR degradation enhancer, ASC-J9®, or neutralization of CCL3 with an antibody, resulted in suppression of macrophage migration and prostate stromal cell growth. Our study provides mechanistic insights on the regulation of prostate stromal cells by macrophages via stromal AR/CCL3 signaling pathways, which could potentially allow the development of therapeutic approaches for battling BPH with persistent inflammation.

Fraction of Macroporous Resin from Smilax china L. Inhibits Testosterone Propionate–Induced Prostatic Hyperplasia in Castrated Rats

The present study was conducted to evaluate the effect of a fraction of macroporous resin (FMR), a bioactive component of Smilax china L., on benign prostatic hyperplasia (BPH) in castrated rats induced by testosterone propionate. Rats were randomly divided into five groups: the negative control group (sham-operated), the model group, two FMR-treated groups (at doses of 300 mg/kg and 600 mg/kg of body weight), and the positive control group (treated with finasteride at the dose of 3 mg/kg). Drugs were administered once a day for three consecutive weeks by gastric gavage. Prostates were weighed, testosterone and dihydrotestosterone (DHT) levels in serum were determined, and histopathological examinations were carried out. FMR treatment inhibited prostatic hyperplasia, reducing the DHT level in serum and improving the prostate gland morphology compared with the model group. The overall results of this study suggest that FMR is effective at inhibiting experimentally induced prostate enlargement, and it presents a valuable resource for the treatment of human BPH.

1570 TADALAFIL, THE PHOSPHODIESTERASE-5 INHIBITOR, ATTENUATES THE CELL GROWTH OF HUMAN BENIGN PROSTATIC HYPERPLASIA XENOGRAFTS IN SUPER-SCID MICE

You have accessJournal of UrologyBenign Prostatic Hyperplasia: Basic Research1 Apr 20121570 TADALAFIL, THE PHOSPHODIESTERASE-5 INHIBITOR, ATTENUATES THE CELL GROWTH OF HUMAN BENIGN PROSTATIC HYPERPLASIA XENOGRAFTS IN SUPER-SCID MICE Hiroshi Kiuchi, Akira Tsujimura, Hidenobu Okuda, Keisuke Yamamoto, Shinichirou Fukuhara, Iwao Yoshioka, Tetsuya Takao, Yasushi Miyagawa, Norio Nonomura, and Taisei Nomura Hiroshi KiuchiHiroshi Kiuchi Suita, Japan More articles by this author , Akira TsujimuraAkira Tsujimura Suita, Japan More articles by this author , Hidenobu OkudaHidenobu Okuda Suita, Japan More articles by this author , Keisuke YamamotoKeisuke Yamamoto Suita, Japan More articles by this author , Shinichirou FukuharaShinichirou Fukuhara Suita, Japan More articles by this author , Iwao YoshiokaIwao Yoshioka Suita, Japan More articles by this author , Tetsuya TakaoTetsuya Takao Suita, Japan More articles by this author , Yasushi MiyagawaYasushi Miyagawa Suita, Japan More articles by this author , Norio NonomuraNorio Nonomura Suita, Japan More articles by this author , and Taisei NomuraTaisei Nomura Ibaraki, Japan More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2012.02.1342AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Benign prostatic hyperplasia (BPH) is characterized by a pathologic proliferation of the prostate glandular and stromal tissues, and subsequent induction of lower urinary tract symptoms. Emerging evidences suggest that phosphodiesterase-5 (PDE-5) inhibitors improve urinary symptoms in patients with BPH. However, inhibition of prostate growth by PDE-5 inhibitors has not been fully elucidated. The objective of this study is to evaluate the effect of Tadalafil, the PDE-5 inhibitor, on suppression of prostate cell growth using a novel human BPH xenograft model. METHODS Human BPH tissue was obtained from a single patient who had undertaken retropubic prostatectomy for the treatment of BPH and was transplanted under the skin of super-SCID mice; SCID mice with macrophage dysfunction. BPH tissues were collected after 2 months of treatment with Tadalafil orally or nothing (control) with sustained release testosterone pellet. Immunohistochemical study was performed to examine the histology and expression of cell type-specific markers in xenografts, including PSA, androgen receptor (AR), p63 (basal cell marker), cytokeratin 8(K8, luminal cell marker) and human-specific CD31 (human vessel marker). We next assess the effect of Tadalafil on the prostate cell proliferation by Ki67 and apoptosis by cleaved caspase-3. Endothelial NOS (eNOS) and COX-2 expression were also examined. RESULTS The implanted tissue contained normal prostatic components; the gland showed two layers of epithelial cells. Inner layer expressed K8, AR and PSA, and outer layer was positive for p63. A considerable amount of the human originated vessels was apparent. After 2 months of Tadalafil treatment, a proliferation index (percentage of Ki-positive cells) was significantly reduced, compared with control in epithelial cells (1.4% vs. 3.9%, p<0.05) as well as in stromal tissues (51.7 vs.14.1 cells/mm2, p<0.05). Cleaved caspase-3 positive cells were significantly increased by Tadalafil. In addition, Tadalafil induced up-regulation of eNOS expression and down-regulation of COX-2 expression. CONCLUSIONS Our novel xenograft model has maintained appropriate histology and protein expression, and may serve as a useful human BPH model. Our study also indicates that Tadalafil attenuates the cell growth and induces apoptosis in benign prostate cells through NO signaling. These results provide a therapeutic potential of Tadalafil for reducing prostate volume in patients with BPH. © 2012 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 187Issue 4SApril 2012Page: e636 Advertisement Copyright & Permissions© 2012 by American Urological Association Education and Research, Inc.MetricsAuthor Information Hiroshi Kiuchi Suita, Japan More articles by this author Akira Tsujimura Suita, Japan More articles by this author Hidenobu Okuda Suita, Japan More articles by this author Keisuke Yamamoto Suita, Japan More articles by this author Shinichirou Fukuhara Suita, Japan More articles by this author Iwao Yoshioka Suita, Japan More articles by this author Tetsuya Takao Suita, Japan More articles by this author Yasushi Miyagawa Suita, Japan More articles by this author Norio Nonomura Suita, Japan More articles by this author Taisei Nomura Ibaraki, Japan More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

11 Marked alterations of gene transcription in the stroma and the epithelium of human benign prostatic hyperplasia

11 Marked alterations of gene transcription in the stroma and the epithelium of human benign prostatic hyperplasia

Modulation of YY1 and p53 expression by transforming growth factor-β3 in prostate cell lines

Transforming growth factor-β (TGF-β) is the prototype of a family of secreted polypeptide growth factors. These cytokines play very important roles during development, as well as in normal physiological and disease processes, by regulating a wide array of cellular processes, such as cell growth, differentiation, migration, apoptosis, and extracellular matrix production. TGF-β utilizes a multitude of intracellular signalling pathways in addition to Smads with actions that are dependent on circumstances, including dose, target cell type, and context. The aims of this research were (i) to verify the effects of dose-dependent TGF-β3 treatment on YY1 and p53 expression, in BPH-1 cell line, human benign prostate hyperplasia, and two prostate cancer cell lines, LNCaP, which is androgen-sensitive, and DU-145, which is androgen-non responsive, (ii) establish a correlation between p53 and YY1 and (iii) determine the expression of a number of important intracellular signalling pathways in TGF-β3-treated prostate cell lines. The expression of YY1, p53, PI3K, AKT, pAKT, PTEN, Bcl-2, Bax, and iNOS was evaluated through Western blot analysis on BPH-1, LNCaP, and DU-145 cultures treated with 10 and 50ng/ml of TGF-β3 for 24h. The production of nitric oxide (NO) was determined by Griess reagent and cell viability through MTT assay. The results of this research demonstrated profound differences in the responses of the BPH-1, LNCaP, and DU-145 cell lines to TGF-β3 stimulation. We believe that the findings could be important because of the clinical relevance that they may assume and the therapeutic implications for TGF-β treatment of prostate cancer.

Intermediate-Conductance Ca2+-Activated K+ Channel, KCa3.1, as a Novel Therapeutic Target for Benign Prostatic Hyperplasia

Recently, a new experimental stromal hyperplasia animal model corresponding to clinical benign prostatic hyperplasia (BPH) was established. The main objective of this study was to elucidate the roles of the intermediate-conductance Ca(2+)-activated K(+) channel (K(Ca)3.1) in the implanted urogenital sinus (UGS) of stromal hyperplasia BPH model rats. Using DNA microarray, real-time polymerase chain reaction, Western blot, and/or immunohistochemical analyses, we identified the expression of K(Ca)3.1 and its transcriptional regulators in implanted UGS of BPH model rats and prostate needle-biopsy samples and surgical prostate specimens of BPH patients. We also examined the in vivo effects of a K(Ca)3.1 blocker, 1-[(2-chlorophenyl)diphenylmethyl]-1H-pyrazole (TRAM-34), on the proliferation index of implanted UGS by measurement of UGS weights and proliferating cell nuclear antigen immunostaining. K(Ca)3.1 genes and proteins were highly expressed in implanted UGS rather than in the normal host prostate. In the implanted UGS, the gene expressions of two transcriptional regulators of K(Ca)3.1, repressor element 1-silencing transcription factor and c-Jun, were significantly down- and up-regulated, and the regulations were correlated negatively or positively with K(Ca)3.1 expression, respectively. Positive signals of K(Ca)3.1 proteins were detected exclusively in stromal cells, whereas they were scarcely immunolocalized to basal cells of the epithelium in implanted UGS. In vivo treatment with TRAM-34 significantly suppressed the increase in implanted UGS weights compared with the decrease in stromal cell components. Moreover, significant levels of K(Ca)3.1 expression were observed in human BPH samples. K(Ca)3.1 blockers may be a novel treatment option for patients suffering from BPH.

Abstract 433: Altered expression of lumican in prostate cancer is identified by microarray analysis

Abstract Prostate cancer is one of the principal medical problems facing men worldwide; fortunately mortality rates from prostate cancer have been decreasing in many developed countries, as a result of improved treatments and early detection. Developing biomarkers that correlate to prostate cancer will facilitate the delivery and development of treatment therapies for patients. To accomplish this we performed a comprehensive genome-wide microarray analysis of genes differentially up- or down-regulated in PSACre;PtenloxP/loxP mutant mice and examined novel differentially expressed genes in to identify novel genes that could potentially serve as biomarkers for disease tumor characterization. Total RNAs were collected form the prostate glands of tumor bearing PSACre;PtenloxP/loxP mutant and wild type mice. cDNAs were generated and were then hybridized onto Affymetrix microarray chips and analyzed with Agilent's Gene Spring software. Microarray analysis revealed that lumican was significantly upregulated in Pten mutant mice compared to wildtype controls. qRT-PCR validation was performed and correlated to the microarray data. Immunohistochemical staining was performed on mouse prostates at various stages of the disease process. Two staining patterns were observed and tabulated, 1) prostate epithelium and 2) stroma. Mouse epithelial cell tended to stain negative in normal glands. Expression increased in moderate and poorly differentiated tumor. The highest expression patterns were observed in the prostates from castrate resistant tumors. More notable was the increased an intense staining seen in the stromal component, especially in the thickened basement membranes cancerous glands. We also examined the expression patterns of lumican in 10 cases of human prostate cancer and benign prostatic hyperplasia (BPH). Epithelial expression in human was moderate to high in benign prostatic hyperplasia, Gleason score 6, 9 and metastatic tumors and moderate in Gleason score 7 tumors. Stromal staining was low in BPH and gradually increased to moderate as the Gleason score increased and high in metastatic lesions. The staining pattern of lumican was very similar to that of vimentin in both human and mouse tissues. Our data has shown that lumican may be differentially expressed in various stages of prostate cancer, however, larger number of clinical need to be evaluated to establish a clear correlation to clinicopathological features. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 433. doi:10.1158/1538-7445.AM2011-433

Expression and significance of estrogen receptor α and β in prostate cancer and peri-cancer tissue

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. Key words: Estrogen receptor alpha; Estrogen receptor beta; Prostatic neoplasms; Carcinoma; Prostatic hyperplasia

Abstract 2167: Identification of aberrant expression of HOXA10 in prostate cancer

Abstract Background: Prostate cancer progression follows a series of defined states, which include prostatic intraepithelial neoplasia (PIN), prostate adenocarcinoma in situ, locally invasive adenocarcinoma and eventually metastatic cancer. One pathway with a prominent role in prostate cancer is the PI3K/Akt/mTOR pathway. Current estimates suggest that PI3K/Akt/mTOR signaling is upregulated in 30-50% of prostate cancers, often through loss of PTEN. In order to identify novel genes potentially regulated by Pten in prostate cancer cells, we performed a comprehensive genome-wide microarray analysis of genes differentially up- or down-regulated in PSACre;PtenloxP/loxP mutant mice. Methods: Total RNAs were collected form the prostate glands of tumor bearing PSACre;PtenloxP/loxP mutant and wild type mice. cDNAs were generated and were then hybridized onto Affymetrix microarray chips and analyzed with Agilent's Gene Spring software. Differentially expressed genes were analyzed and candidate genes were selected and validated by qRT-PCR. Protein expression levels in mouse prostates were analyzed using western blot and immunohistochemical methods. Protein expression levels in human tissues were determined by immunohistochemical methods in archived formalin fixed paraffin embedded tissues. Results: Microarray analysis revealed that HOXA10 was significantly downregulated in Pten mutant mice compared to wildtype controls. qRT-PCR results from mice at different stage of tumor development revealed a different pattern of expression. HOXA10 is a member of the homeobox gene family and is a transcription factors primarily involved in embryonic development. Therefore, we examined the expression of HOXA10 at the protein level by immunohistochemical staining of mouse prostate cancer ranging from normal to highly metastatic cancer. Or observations revealed that expression is high in normal and hyperplastic glands and gradually decreases as tumor grade increases. However, HOXA10 expression rose focally in poorly differentiated tumors and was highly expressed in metastatic tumors. We examined the expression patterns of HOXA10 in 10 cases of human prostate cancer and benign prostatic hyperplasia. As in mice, expression was high in normal and benign glands; moderate in Gleason score 6; low in Gleason score 7-8; low with nest of moderate to strong stained cells in Gleason score 9; and high in metastasis. Immunohistochemical hierarchical cluster analysis showed an inverse pattern of expression between HOXA10 and vimentin. Conclusion: Our results have shown that HOXA10 is aberrantly expressed in both human and mouse prostate cancer and appears to be involved in the carcinogenesis process, particularly in the development of metastasis. Further work is currently ongoing to elucidate the role HOXA10 plays in prostate cancer. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2167. doi:10.1158/1538-7445.AM2011-2167

A cyclooxygenase-2 inhibitor reduces serum prostatic-specific antigen levels in men with benign prostatic hyperplasia and lower urinary tract symptoms and helps differentiate chronic inflammation from prostate cancer

Emerging evidence indicates that prostatic inflammation may contribute to prostatic growth in benign prostatic hyperplasia (BPH) and neoplastic changes (prostate cancer). The Medical Therapy of Prostatic Symptoms study showed that men with inflammation had a significantly higher risk of BPH progression and acute urinary retention. Evidence also shows that a cyclooxygenase-2 (COX-2) inhibitor can increase apoptotic activity in human BPH tissue. COX-2 inhibitors in combination with alpha-adrenergic blockers may increase the effectiveness of therapy for lower urinary tract symptoms (LUTS) secondary to BPH without significant side effects. COX-2 inhibitors may decrease the serum prostatic-specific antigen level and act as a biomarker to differentiate chronic inflammation from prostate cancer in patients with LUTS suggestive of BPH who do not have a palpable prostate nodule but have a serum prostatic-specific antigen level higher than 4 ng/mL. The results might provide a simple way to initially differentiate chronic inflammation from prostatic cancer. If chronic inflammation of the prostate can be reduced by COX-2 inhibitor treatment, bothersome LUTS as well as voiding conditions might be improved.

Proteomics-based signature for human benign prostate hyperplasia and prostate adenocarcinoma

Prostate adenocarcinoma often presents at a late stage, due to a lack of early clinical symptoms and lack of accurate objective markers. This study aimed to identify and validate proteomics-based biomarkers useful for prostate cancer diagnosis and to establish a marker-panel for prostate cancer and benign prostate hyperplasia (BPH). Global protein expression patterns in fresh tissue specimens from 8 patients with prostate carcinoma and 16 with BPH were analyzed by two-dimensional gel electrophoresis. Differentially expressed proteins were identified by MALDI-TOF mass spectrometry. We compared our results with those of published studies and defined a set of common biomarkers. We identified 22 differentially expressed proteins between BPH and prostate carcinomas. The up-regulated proteins in cancer compared to BPH included protein disulfide-isomerase, 14-3-3-protein, Enoyl CoA-hydrase, prohibitin and B-tubulin β-2. Keratin-II, desmin, HSP71, ATP-synthase-β-chain and creatine kinase-β-chain were down-regulated. Survey of the literature showed that 15 of our 22 identified proteins have been previously reported to differ in their expression levels between BPH and prostate cancer by other laboratories. The expression patterns of these biomarkers could successfully cluster BPH and adenocarcinomas as well as prostate cancer of low and high Gleason scores. This study validates protein-biomarkers that can be useful for accurate diagnosis and prognostic monitoring of prostate adenocarcinoma. Despite varied prevalence of the disease between different ethnic populations (i.e., high in Sweden, low in Saudi Arabia); the biomarkers indicate that BPH and prostate cancers are biologically 'homogeneous' in their protein expression patterns across wide geographical regions.

Expression of leukemia/lymphoma related factor (LRF/Pokemon) in human benign prostate hyperplasia and prostate cancer

Leukemia/lymphoma related factor (LRF), also known as Pokemon, is a protein that belongs to the POK family of transcriptional repressors. It has an oncogenic role in many different solid tumors. In this study, the expression of LRF was evaluated in benign prostate hyperplastic (BPH) and prostate cancer (PC) tissues. The functional expression of LRF was studied using multiple cellular and molecular methods including RT-PCR, western blotting, immunohistochemistry, and immunofluorescence. Paraffin-embedded human tissues of BPH and PC were used to examine LRF expression. Histological staining of the BPH and PC tissue sections revealed nuclear expression of LRF with minimal expression in the surrounding stroma. The semi-quantitative RT-PCR and western immunoblot analyses demonstrated significantly higher mRNA transcripts and protein expression in PC than BPH. High expression of LRF suggests that it may have a potential role in the pathogenesis of both BPH and prostate cancer. Further studies will help elucidate the mechanisms and signaling pathways that LRF may follow in the pathogenesis of prostate carcinoma.

Selective estrogen receptor modulators regulate stromal proliferation in human benign prostatic hyperplasia by multiple beneficial mechanisms—action of two new agents

The existing drugs for benign prostatic hyperplasia (BPH) are partially effective with undesirable side-effects; hence new agents acting by different mechanism(s) are required as supplements. Modulation of estrogen receptor signaling using selective estrogen receptor modulators (SERMs) offers an alternative approach for BPH management. Using human BPH-derived stromal cells and tissue explants in culture we evaluated two SERMs, DL-2-[4-(2-piperidinoethoxy)phenyl]-3-phenyl-2 H-1-benzopyran (BP) and Ormeloxifene (Orm) in comparison to Tamoxifen (Tam) and 4-hydroxytamoxifen (OHT). BP, OHT and Tam were more effective than Orm in reducing stromal cell proliferation of human BPH. BP was either equipotent or more effective than OHT and Tam in increasing estrogen receptor(ER)-ß, TGFß1, Fas and FasL, and in decreasing ER-α, AR, EGF-R and IGF-I expressions in BPH stromal cells. BP, Tam and Orm (1.0 mg/Kg) reduced rat prostate weights by almost same extent as Finasteride (Fin, 5.0 mg/Kg); however combination treatment (SERM+Fin) was more effective. BP was exceptionally efficient in reducing IGF-1 and cleaving PARP while combination treatments more effectively increased bax:bcl-2 ratio. Fin reduced acinar diameter and prostatic DHT level but increased testosterone, estradiol (E(2)) and E(2)/T+DHT ratio. SERMs, especially BP, reduced epithelial cell height drastically without significantly altering steroid hormone levels and E(2)/T+DHT ratio. Combination treatment reduced both acinar diameter and epithelial cell height with modest increase in E(2), T and E(2)/T+DHT. The study reveals the potential of SERMs per se for BPH management, and more effectively in combination with a 5α-reductase inhibitor. BP appears promising for further evaluation as a drug candidate for BPH and prostate cancer.

Abstract B46: The effect of sulforaphane on protein expression in benign prostatic hyperplasia tissue

Abstract Background: Benign prostatic hyperplasia (BPH) is a common condition that occurs in over 25% of men aged 50+, increasing to 50% amongst those over 80. Although the etiology of the condition has not been ascertained, the majority of BPH specimens contain inflammatory infiltrates indicating inflammation plays an important role in the progression of the disease. Furthermore, a role for inflammation has also been suggested in prostate cancer (PCa). BPH is not a pre-cursor of PCa but using BPH as a model of inflammation could result in findings that may apply and add to our understanding of PCa. Previous work has demonstrated that the consumption of broccoli can alter inflammatory pathways in the human prostate, and this may be due to the action of the isothiocyanate sulforaphane (SF), a phytochemical derived from broccoli. This dietary compound has a range of biological activities, including reducing inflammation. Hypothesis: Sulforaphane reduces inflammatory pathways and beneficially alters protein expression in BPH tissue. Methods: We obtained and cultured human BPH tissue for 24 hours with or without 10µM SF and collected tissue and media for further analysis using two distinct approaches. Firstly, in a targeted approach, known inflammatory markers were measured in the media of five tissue samples by commercially available ELISA. Secondly, a global proteomic analysis was carried out to give an unbiased view of the effect of SF on the protein expression within the tissue. Initially, protein was extracted from the tissue of three individuals, run on 2D gels and spots of interest excised from the gel and in-gel trypsin digested. The resulting peptides were analyzed on an LTQ-Orbitrap Mass Spectrometer and the proteins identified by Mascot. Selected proteins were then further quantified in additional tissue samples. Results: Analysis of the effect of SF on inflammatory markers revealed that SF significantly reduced levels of secreted IL-6 and to a lesser extent IL-8 and FGF-2. Global proteomic analysis demonstrated that there was, as expected, considerable variation between individuals, and that SF altered expression of several proteins in BPH tissue. Reduction of GRP94, an endoplasmic reticulum chaperone protein observed in one individual, was further verified in independent BPH samples. Initial immunohistochemistry in sections of BPH tissue suggests that SF reduces the nuclear localization of this protein. Further experiments are currently ongoing to investigate these effects in more individuals and also to look at the biological consequences. Conclusion: Natural variation present within humans highlights the need to use a larger number of samples in order to understand the complex effects of SF on prostate tissue. The results suggest that SF can reduce inflammatory markers and alter proteins within the human prostate indicating a possible protective role for SF in pathogenesis of the prostate. Citation Information: Cancer Prev Res 2010;3(12 Suppl):B46.

Receptors for luteinizing hormone‐releasing hormone (LHRH) in benign prostatic hyperplasia (BPH) as potential molecular targets for therapy with LHRH antagonist cetrorelix

The majority of men will develop symptoms of benign prostatic hyperplasia (BPH) after 70 years of age. Various studies indicate that antagonists of LHRH, such as cetrorelix, exert direct inhibitory effects on BPH mediated by specific LHRH receptors. Our aim was to investigate the mRNA for LHRH and LHRH receptors and the expression of LHRH receptors in specimens of human BPH. The expression of mRNA for LHRH (n=35) and LHRH receptors (n=55) was investigated by RT-PCR in surgical specimens of BPH, using specific primers. The characteristics of binding sites for LHRH on 20 samples were determined by ligand competition assays. The LHRH receptor expression was also examined in 64 BPH specimens by immunohistochemistry. PCR products for LHRH were found in 18 of 35 (51%) BPH tissues and mRNA for LHRH receptors was detected in 39 of 55 (71%) BPH specimens. Eighteen of 20 (90%) samples showed a single class of high affinity binding sites for [D-Trp(6) ]LHRH with a mean K(d) of 4.04 nM and a mean B(max) of 527.6 fmol/mg membrane protein. LHRH antagonist cetrorelix showed high affinity binding to LHRH receptors in BPH. Positive immunohistochemical reaction for LHRH receptors was present in 42 of 64 (67%) BPH specimens. A high incidence of LHRH receptors in BPH supports the use of LHRH antagonists such as cetrorelix, for treatment of patients with lower urinary tract symptoms from BPH.