Benign Prostatic Hyperplasia Cells Research Articles

Effect of Urinary Retention on the Satisfaction and Complication Rate in Benign Prostatic Hyperplasia Patients Undergoing Prostatectomy

A cohort study from January 2000 till January 2005 included all patients who underwent prostatic surgery with histopathology report of benign prostatic hyperplasia. Patients were divided into elective or retention group. Data of patient demographics, satisfaction and postoperative complications were collected. Total of 119 patients, retention (n = 30), elective group (n = 89). Retention rate was 25%. There was no eff ect of retention on postoperative complication (elective = 44%, retention = 41%) p value = 0.826, odds ratio 0.878, CI (0.363 - 2.124) nor any eff ect on patients' satisfaction (elective = 54%, retention = 59%) p value = 0.661, odds ration = 1.256, CI (0.520 - 3.034). Patients’ age and prostate size did not show any effect on postoperative outcome. While presence of infl ammatory cells in benign prostatic hyperplasia (BPH) histopathology showed positiveeff ect on satisfaction (BPH alone = 47%, BPH + infl ammation = 71%) p value = 0.037, and a protective eff ect on postoperative complications (BPH alone = 45%, +BPH with infl ammation = 18%) p value = 0.167. Conclusion: Retention rate is comparable to the international reports. The presence of infl ammatory cells in the benign prostatic hyperplasia histopathology showed a positive eff ect on postoperative satisfaction.

Acute and subacute toxicity of an ethanolic extract of Melandrii Herba in Crl:CD sprague dawley rats and cytotoxicity of the extract in vitro.

BackgroundMelandrii Herba, a medicinal plant, has been used in Korea for treatment of bacterial and fungal infection. However, the safety and toxicity of Melandrii Herba have not yet been established. Therefore, we investigated the acute and subacute toxicity of an ethanolic extract of Melandrii Herba (MHEE) in Crl:CD Sprague Dawley rats and cytotoxicity of MHEE in vitro.MethodsTo study acute toxicity, rats were treated with MHEE at single doses of 0, 500, 1000, and 2000 mg/kg administered by oral gavage, and body weight, clinical signs, and mortality were observed after dosing. To study subacute toxicity, rats were treated with MHEE at doses of 0, 500, 1000, and 2000 mg/kg administered once a day by gavage for 4 weeks. We measured clinical signs, mortality, gross pathological findings, body and organ weights, food consumption, serum biochemistry, and conducted hematology and urinalysis. The cytotoxicity of MHEE was assayed by measuring the viability of prostate cell lines including normal prostate stromal WPMY-1, normal prostate epithelial RWPE-1, and benign prostatic hyperplasia epithelial BPH-1 cells at various concentrations of MHEE in vitro.ResultsSingle oral doses of MHEE caused no significant difference in rat clinical signs, mortality, or body weight. The lethal dose of MHEE was considered to be >2000 mg/kg. Daily oral doses of MHEE for 4 weeks did not result in any significant changes in rat mortality, gross pathological findings, relative organ weights, food consumption, hematology, serum biochemistry, or urinalysis. At MHEE >1000 mg/kg/day, salivation was increased in both male and female rats. However, the salivation caused by the MHEE treatment was not accompanied by pathological changes in body weight or gross pathological findings, and we considered the salivation as a minor symptom. Therefore, no adverse effects were seen at 2000 mg/kg/day or less. MHEE showed no cytotoxic effects on either normal prostate or benign prostatic hyperplasia cell lines.ConclusionsAdministration of MHEE in Crl:CD Spradgue Dawley rats is nontoxic and is safe for at least a month.

Chemosensitization of Prostate Carcinoma Cells with a Receptor-directed Smac Conjugate.

Second mitochondrial activator of caspase (Smac) is a short mitochondrial peptide. When released from the mitochondria into the cytoplasm, it binds to inhibitor of apoptotic proteins (IAPs) within the cytoplasm and prevents them from inhibiting apoptosis. Delivery of external synthetic Smac peptide into the cytoplasm of malignant cells could greatly improve the efficiency of apoptosis-inducing chemotherapeutic agents. In our study different conjugates based on the seven N-terminal amino acids AVPIAQK of Smac (SmacN7) were produced to obtain a cytoplasm-directed Smac variant. SmacN7 and a point mutant (AVPKAQK) were coupled either to rhodamine alone or to both rhodamine and undecylic aldehyde, which is an antagonist of the Lily-of-the-valley fragrance receptor. The fifth conjugate consisted of rhodamine coupled only to undecylic aldehyde, without SmacN7. The uptake of these five conjugates into three different human cell lines was characterized and quantified by confocal laser scanning microscopy and flow cytometry. A caspase apoptosis assay was performed for cells incubated with the five different conjugates after induction of apoptosis. The coupling of undecylic aldehyde to SmacN7 increased the cellular uptake of the correct and mutant conjugates. Caspase 3/7 apoptosis tests after induction of apoptosis with staurosporine or UV irradiation showed that the coupling of SmacN7 with undecylic aldehyde resulted in a greatly increased adjuvant pro-apoptotic effect compared to the separate components and a mutant SmacN7 peptide sequence in the LNCaP prostate carcinoma cells compared to the benign prostate hyperplasia (BPH) cells and the human embryonal kidney (HEK) cells.

SU‐F‐SPS‐08: Measuring the Interaction Of DDR Cell Receptors and Extracellular Matrix Collagen in Prostate Cells

Purpose:Discoidin domain receptors (DDR) have recently been recognized as important players in cancer progression. DDRs are cell receptors that interact with collagen, an extracellular matrix (ECM) protein. However the detailed mechanism of their interaction is unclear. Here we attempted to examine their interaction in terms of structural (surface topography), mechanical (rupture force), and kinetic (binding probability) information on the single molecular scale with the use of atomic force microscopy (AFM).Methods:The Quantitative Nano‐mechanical property Mapping (QNM) mode of AFM allowed to assess the cells in liquid growth media at their optimal physiological while being viable. Human benign prostate hyperplasia (BPH‐1) cell line was genetically regulated to suppress DDR expression (DDR‐ cells) and was compared with naturally DDR expressing cells (DDR+).Results:Binding force measurements (n = 1000) were obtained before and after the two groups were treated with fibronectin (FN), an integrin‐inhibiting antibody to block the binding of integrin. The quantification indicates that cells containing DDR bind with collagen at a most probable force of 80.3–83.0 ±7.6 pN. The probability of them binding is 0.167 when other interactions (mainly due to integrin‐collagen binding) are minimized.Conclusion:Together with further force measurements at different pulling speeds will determine dissociation rate, binding distance and activation barrier. These parameters in benign cells provides some groundwork in understanding DDR's behavior in various cell microenvironments such as in malignant tumor cells.Funding supported by Richard Barber Interdisciplinary Research Program of Wayne State University

Regulation of Prostate Development and Benign Prostatic Hyperplasia by Autocrine Cholinergic Signaling via Maintaining the Epithelial Progenitor Cells in Proliferating Status.

SummaryRegulation of prostate epithelial progenitor cells is important in prostate development and prostate diseases. Our previous study demonstrated a function of autocrine cholinergic signaling (ACS) in promoting prostate cancer growth and castration resistance. However, whether or not such ACS also plays a role in prostate development is unknown. Here, we report that ACS promoted the proliferation and inhibited the differentiation of prostate epithelial progenitor cells in organotypic cultures. These results were confirmed by ex vivo lineage tracing assays and in vivo renal capsule recombination assays. Moreover, we found that M3 cholinergic receptor (CHRM3) was upregulated in a large subset of benign prostatic hyperplasia (BPH) tissues compared with normal tissues. Activation of CHRM3 also promoted the proliferation of BPH cells. Together, our findings identify a role of ACS in maintaining prostate epithelial progenitor cells in the proliferating state, and blockade of ACS may have clinical implications for the management of BPH.

Circulating Prostate Gland Cells in Benign Prostatic Hyperplasia and Adenocarcinoma in Patients of African Heritage: Diagnosis by Liquid Biopsy: Preliminary Results

Circulating Prostate Gland Cells in Benign Prostatic Hyperplasia and Adenocarcinoma in Patients of African Heritage: Diagnosis by Liquid Biopsy: Preliminary Results

New Multi-target Antagonists of α1A-, α1D-Adrenoceptors and 5-HT1A Receptors Reduce Human Hyperplastic Prostate Cell Growth and the Increase of Intraurethral Pressure.

Benign prostatic hyperplasia (BPH) is characterized by stromal cell proliferation and contraction of the periurethral smooth muscle, causing lower urinary tract symptoms. Current BPH treatment, based on monotherapy with α1A-adrenoceptor antagonists, is helpful for many patients, but insufficient for others, and recent reports suggest that stimulation of α1D-adrenoceptors and 5-hydroxytryptamine (serotonin) (5-HT)1A receptors contributes to cell proliferation. In this study, we investigated the potential of three N-phenylpiperazine derivatives (LDT3, LDT5, and LDT8) as multi-target antagonists of BPH-associated receptors. The affinity and efficacy of LDTs were estimated in isometric contraction and competition-binding assays using tissues (prostate and aorta) and brain membrane samples enriched in specific on- or off-target receptors. LDTs' potency was estimated in intracellular Ca(2+) elevation assays using cells overexpressing human α1-adrenoceptor subtypes. The antiproliferative effect of LDTs on prostate cells from BPH patients was evaluated by viable cell counting and 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide assays. We also determined LDTs' effects on rat intraurethral and arterial pressure. LDT3 and LDT5 are potent antagonists of α1A-, α1D-adrenoceptors, and 5-HT1A receptors (Ki values in the nanomolar range), and fully inhibited phenylephrine- and 5-HT-induced proliferation of BPH cells. In vivo, LDT3 and LDT5 fully blocked the increase of intraurethral pressure (IUP) induced by phenylephrine at doses (ED50 of 0.15 and 0.09 μg.kg(-1), respectively) without effect on basal mean blood pressure. LDT3 and LDT5 are multi-target antagonists of key receptors in BPH, and are capable of triggering both prostate muscle relaxation and human hyperplastic prostate cell growth inhibition in vitro. Thus, LDT3 and LDT5 represent potential new lead compounds for BPH treatment.

Study of Response to Zinc in Human Prostate Spheroids Using X-Ray Microfluorescence

Diseases of the prostate gland are the most frequently diagnosed in men after middle age. Spheroid cell culture is a useful technique for tissue engineering or regenerative medicine research, pharmacological and toxicological studies, and fundamental studies in cell biology. In this paper, we investigated zinc distribution in cell spheroids in benign prostate hyperplasia (BPH) and prostate cancer (PCa) (DU145) and analyzed the differences in the response to zinc (0–150 $\mu \text{M}$ ) treatment using $\mu $ XRF with synchrotron radiation. The results showed nonuniform distribution of Zn in all the spheroids analyzed. The differential response to zinc of BPH and DU145 cell spheroids suggests that zinc may have an important role in PCa.

Abstract 223: Generation of single cell-derived normal, benign and cancer mini-prostates from primary patient-derived tissues

Abstract Clinically effective next generation androgen deprivation therapy (ADT) and androgen receptor (AR)-directed therapies have been developed to treat castration-resistant prostate cancer (CRPC). However, disease progression inevitably occurs, and majority of relapsed tumors continue to overexpress AR. AR mutations and constitutively active AR splice variants (AR-Vs) are detected during ADT and may lead to drug resistance. Therefore, there is dire need to develop models that provide a better understanding of AR signaling and determine markers of ADT resistance in prostate cancer patient cells. Mouse models and existing human prostate cancer cell lines have inherent limitations for deciphering mechanisms of therapy resistance. Unlike the recently reported prostate tissue organoids that were derived at a low efficiency from 7/49 metastatic prostate cancer samples, we have developed a novel and exciting technology to generate normal, benign prostatic hyperplasia (BPH) and cancer “mini-prostates” from both early-stage and advanced prostate cancer patient samples. We have successfully rederived mini-prostates from 17 out of 19 primary prostatectomy tissues. We incorporated developmental epithelial and mesenchymal signals, stem-like cells and growth requirements in 3D cultures for maintaining primary normal, BPH and prostate cancer patient-derived cells as stem-like cell-derived organoids. Our preliminary data show the establishment of patient- single cell-derived normal, BPH and cancer mini-prostates comprised of organoids with glandular tissues that have a thick basement membrane, an outer mesenchymal and inner epithelial cells conveying AR signaling, expressing cytokeratins and secreting PSA. RNA sequencing, expression profiling and ADT revealed that mini-prostate cells maintain the genetic, chemoresistance, and growth rate features of the primary prostate. To model tumor heterogeneity, prostate cells from surgical tissues were isolated upon digital imaging and tissue mapping in mirror sections for AMACR, AR, AR-Vs, p63, Ki67, ERG expression and FISH analysis for TMPRSS2-ERG fusion. Living cells from mapped BPH and cancer clones are being utilized to derive single cells into mini-prostates. Our studies generated novel therapy sensitivity and drug discovery models that would allow for precision medicine and coclinical approaches to be enlisted to reveal chemo- or hormonal-resistance profiles in genetically defined mini-prostates, apply combinatory-targeted therapies and study therapy-induced clonal selection processes to ultimately guide future clinical trials. Citation Format: Monica Bartucci, Michele Patrizii, Eric Huselid, Shamila Yussuf, Nitu Bansal, Kathleen Flaherty, Denis Tolkunov, Hua Zhong, Mark N. Stein, Joseph Bertino, Robert DiPaola, Isaac Kim, Hatem E. Sabaawy. Generation of single cell-derived normal, benign and cancer mini-prostates from primary patient-derived tissues. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 223. doi:10.1158/1538-7445.AM2015-223

The GnRH Antagonist Degarelix Directly Inhibits Benign Prostate Hyperplasia Cell Growth.

Gonadotropin-releasing hormone receptors (GnRHR) have been found in extrapituitary tissues, including the prostate, where they might exert a local effect on tissue growth. Degarelix is a GnRHR antagonist approved for use in patients with prostate cancer (PCa) who need androgen deprivation therapy. The slowing of prostate cell growth is a common goal shared by PCa and benign prostate hyperplasia (BPH) patients, and the effect of degarelix on BPH cells has not yet been investigated. We wanted to evaluate the direct effect of degarelix on human BPH primary cell growth. Gene expression studies performed with BPH (n=11), stage 0 (n=15), and PCa (n=65) human specimens demonstrated the presence of GNRHR1 and GNRHR2 and their respective endogenous peptide ligands. BPH-isolated epithelial and stromal cells were either cultured alone or co-cultured (1:4 or 4:1 ratio of epithelial to stromal cells) and subsequently treated with increasing concentrations of degarelix. Degarelix treatment induced a decrease in cell viability and cell proliferation rates, which occurred in parallel to an increase in apoptosis. Both epithelial and stromal BPH cells are sensitive to degarelix treatment and, interestingly, degarelix is also effective when the cells were growing in a co-culture microenvironment. In contrast to degarelix, the GnRHR agonists, leuprolide and goserelin, exerted no effect on the viability of BPH epithelial or stromal cells. In conclusion, (i) prostate tissues express GNRHR and are a potential target for degarelix; and (ii) degarelix directly inhibits BPH cell growth through a decrease in cell proliferation and an increase in apoptosis. Supporting information for this article is available online at http://www.thieme-connect.de/products.

MP31-08 A TISSUE SPECIFIC ROLE FOR LIGAND INDEPENDENT ARV7 SIGNALING IN BENIGN PROSTATE PATHOGENESIS

You have accessJournal of UrologyBenign Prostatic Hyperplasia: Basic Research1 Apr 2015MP31-08 A TISSUE SPECIFIC ROLE FOR LIGAND INDEPENDENT ARV7 SIGNALING IN BENIGN PROSTATE PATHOGENESIS Tyler M. Bauman, Emily A. Ricke, Wei Huang, and William A. Ricke Tyler M. BaumanTyler M. Bauman More articles by this author , Emily A. RickeEmily A. Ricke More articles by this author , Wei HuangWei Huang More articles by this author , and William A. RickeWilliam A. Ricke More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2015.02.1363AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Androgen receptor variant 7 (ARv7) is a splice variant of the androgen receptor (AR) that is thought to act independent of androgens due to the absence of a ligand-binding domain. ARv7 is an important mediator of castration-resistant prostate cancer (CRPC), but the role of ARv7 in benign prostate pathology has not yet been elucidated. We hypothesized that protein expression of ARv7 would be increased in benign prostatic hyperplasia (BPH) compared to normal prostate tissue, representing a mechanism of androgen-independent growth in BPH. METHODS Multiplexed immunohistochemistry was performed using antibodies for full-length AR and ARv7 on a tissue microarray containing tumor-adjacent normal prostate (normal; n=96 cores from 48 patients) and BPH (n=48 cores from 24 patients), and an automated pathology platform was used to quantify protein expression, positivity, and co-localization. Epithelial-specific and stromal-specific expression and co-localization of AR and ARv7 were compared between BPH and normal prostate. RESULTS AR and ARv7 were observed in nuclei of stromal and epithelial cell from all normal and pathologic specimens. In epithelial tissue, nuclear expression and positivity of both ARv7 and AR were significantly increased in BPH compared to normal prostate tissue (p<0.0001). The proportion of double negative (AR-/ARv7-) cells was lower in BPH than normal prostate (p<0.0001), and the number of double positive (AR+/ARv7+) cells was increased in BPH (p<0.0001). In the stroma, nuclear expression (p<0.0001) and positivity (p=0.0009) of ARv7 were significantly increased in BPH compared to normal prostate tissue. AR expression and positivity were also higher in BPH (p<0.0001). The proportion of AR-/ARv7+ single positive cells was increased in BPH (p=0.006), as was the proportion of AR+/ARv7- single positive cells (p=0.002). AR+/ARv7+ double positive cells were three-fold higher in BPH compared to normal prostate (p<0.0001), and the proportion of AR-/ARv7- double negative cells was decreased in BPH (p<0.0001). The ratio of ARv7:AR was lower in BPH than normal prostate tissue (p=0.02) for both tissue compartments. CONCLUSIONS Both epithelial and stromal cells in BPH express higher levels of ARv7 than normal prostate tissue. Considerable increases in ARv7/AR nuclear co-localization in BPH may represent a novel mechanism driving hyperplasia. Taken together, these results underscore the potentiation for androgen-independent growth in BPH and may have significant consequences for men treated with 5α-reductase inhibitors that fail therapy. © 2015 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 193Issue 4SApril 2015Page: e358 Advertisement Copyright & Permissions© 2015 by American Urological Association Education and Research, Inc.MetricsAuthor Information Tyler M. Bauman More articles by this author Emily A. Ricke More articles by this author Wei Huang More articles by this author William A. Ricke More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

793 The role of macrophage migration inhibitory factor (MIF) in benign prostatic hyperplasia (BPH): Promote BPH cell proliferation via modulation of COX-2 and p53 signaling

793 The role of macrophage migration inhibitory factor (MIF) in benign prostatic hyperplasia (BPH): Promote BPH cell proliferation via modulation of COX-2 and p53 signaling

Study of the differential response to zinc of prostate cell spheroids using the synchrotron radiation X‐ray microfluorescence technique

X‐ray microfluorescence imaging technique has very often been used as a significant tool in biomedical research to obtain information regarding the distribution of elements in tissue and cell samples. This study investigated Zn distribution in benign prostate hyperplasia (BPH) and prostate cancer (DU145) cell spheroids and analyzed the different response to Zinc treatment using synchrotron X‐ray microfluorescence (μSRXRF). The μSRXRF measurements were performed at the XRF beam line at the Synchrotron Light National Laboratory (Campinas, Brazil). The results showed non‐uniform distribution of Zn in all the spheroids analyzed. The differential response of DU145 and BPH cell spheroids to zinc suggests that there are different mechanisms of Zn accumulation in tumor cells. Copyright © 2015 John Wiley &amp; Sons, Ltd.

DLEC1, a 3p tumor suppressor, represses NF-κB signaling and is methylated in prostate cancer.

Deleted in lung and esophageal cancer 1 (DLEC1), located at 3p22-p21.3, is involved in the carcinogenesis of multiple cancers, but its role in prostate cancer (PrCa) remains unclear. Here, we studied the epigenetic alteration of DLEC1 and its functions in prostate cancer. We found that DLEC1 was highly expressed in normal prostate tissues, normal prostatic epithelium cell line (RWPE-1), and benign prostatic hyperplasia cell line (BPH-1), but frequently downregulated by promoter methylation in PrCa cell lines. Pharmacologic demethylation could restore DLEC1 expression. DLEC1 was downregulated in prostate tumor tissues compared with their adjacent non-malignant tissues. DLEC1 was methylated in 76/110 primary tumors, but rarely in benign prostatic hyperplasia tissues. DLEC1 methylation was associated with higher PSA levels (p = 0.016), higher Gleason scores (p = 0.015), and more advanced tumor stages (p = 0.003). Furthermore, DLEC1 methylation was detected in 11/30 urine sediment samples from PrCa patients, but seldom in ones from BPH patients. Ectopic expression of DLEC1 inhibited the colony formation of PrCa cells, through inducing cell apoptosis. DLEC1 also suppressed PrCa cell migration. Moreover, DLEC1 inhibited NF-κB transcription activity in PrCa and HEK293 cells. Taken together, our data demonstrate that DLEC1 functions as a tumor suppressor but is frequently methylated in prostate cancer. DLEC1 methylation is associated with prostate cancer progression, which could be a non-invasive epigenetic biomarker for PrCa diagnosis. • Promoter methylation of DLEC1 is a potential prognostic biomarker for PrCa. • DLEC1, a functional tumor suppressor, is frequently methylated in PrCa. • DLEC1 suppresses prostate cancer growth and metastatic behavior. • DLEC1 mediates tumor-suppressive activities through NF-κB signaling.

The Differential Expression and Function of the Inflammatory Chemokine Receptor CXCR5 in Benign Prostatic Hyperplasia and Prostate Cancer.

Background: Chemokine and chemokine receptors could have played an important role in tumor angiogenesis and distant metastasis. The mechanism of inflammation, expression and function of chemokines and chemokine receptors in benign prostatic hyperplasia (BPH) and prostate cancer (PCa) remain unclear. The purpose of present study is to detect differential expression and function of chemokines and chemokine receptors (CCRs) in BPH and PCa.Methods: BPH-1 and peripheral blood mononuclear cells (PBMCs) were co-cultured in Transwell chambers, and human normal prostate (NP) tissues, BPH tissues and PCa tissues were collected. CCR gene-chips were used to analyze and compare the differential expression of CCRs in BPH-1 cells, BPH-1 cells co-cultured with PBMCs, and LNCaP cells. The differential expression of CCRs was detected and validated using real-time PCR, western blotting and immunofluorescence (IF). The proliferation of LNCaP cells was also investigated after the knockdown CXCR5.Results: Results of gene-chips indicated that there was low or no expression of CCR10, CXCR1, CXCR3 and CXCR5 in BPH-1 cells, whereas the expression of these receptors in BPH-1 cells was increased by PBMCs, and the expression was high in LNCaP cells. Furthermore, real-time PCR and western blotting confirmed the above mentioned results. IF verified no or low expression of CXCR1, CXCR3 and CXCR5 in NP tissues, low or moderate expression in BPH and high expression in PCa. However, CCR10 was not expressed at detectable levels in the three groups. The growth and proliferation of LNCaP cells was markedly inhibited after down-regulation of CXCR5.Conclusions: PCa cells expressed high levels of CCR10, CXCR1, CXCR3 and CXCR5. Although BPH cells did not express these factors, their expression was up-regulated when BPH-1 cells were incubated with inflammatory cells. Finally, down-regulation of CXCR5 inhibited the growth and proliferation of LNCaP cells.

Abstract 3432: A genome-wide shRNA screen for suppressors of prostate cancer cell invasion

Abstract Metastasis in prostate cancer is caused by genetic reprogramming leading to increased cell motility and the formation of invasive structures which allow cells to invade out of the primary tumour in a process known as intravasation. In order to search for novel genes inhibiting invasive phenotypes responsible for intravasation, a genome-wide high throughput short hairpin RNA screen was used to identify genes which suppress aggressive phenotypes in BPH (Benign Prostatic Hyperplasia) prostate epithelial cells when grown in 3D matrigel culture conditions. Using the The RNAi Consortium (TRC) lentiviral shRNA library that covers the entire human transcriptome (80,000 unique constructs), BPH cells were infected such that each cell would integrate a single shRNA construct that will target mRNA expression of a known human gene. Cells were plated at 4500 cells per cm2 until a 2.5X genome-wide coverage was achieved. In 3D matrigel culture conditions, BPH colonies exhibit normal epithelial (round) colony morphology. Hence, we screened for clonogenic BPH colonies that appeared fibroblastic and spindle-shaped. Collection of these “hits” yield gene identities that suppress intravasation. In this ongoing study, we have achieved 2.5x coverage of the TRC shRNA library in BPH cells. After 11 days of growth, 7 “hits” from the BPH screen that exhibit an invasive phenotype were isolated, and were validated again in 3D matrigel culture. These “hits” have revealed 12 potential invasion suppressors involved in pathways including ER stress, cortactin phosphorylation signalling, cell differentiation and genome stability. These hits are currently being re-validated with other shRNAs specific for each gene. Overall, we provide a high-throughput and high-content screen for discovering novel genes that inhibit prostate cancer progression. These functional genomics screens which focus on altered prostate cell colony phenotypes may open new doors for potential drug treatments and a greater understanding of prostate cancer and BPH. Genome-wide shRNA screen invasion suppressor genesHit #GeneSymbolCellular Function1Interleukin 19IL19Upregulates IL6 and TNFA1Polyhomeotic-like protein 3PHC3Part of polycomb group complex (repressor)2Zinc finger protein 397ZNF397Repressor/activator transcription factor2Transmembrane protein 189-ubiquitin-cojugating enzyme E2 variant 1TMEM189-UBE2V1Cytosolic ubiquitination protein3Nuclear protein localization 4 homologNPLOC4Involved in ER stress (retro-translocation of proteins for degradation)4Flavin adenine dinucleotide synthetase 1FADSConverts FMN to FADH25Selenoprotein SSELSInvolved in ER stress (retro-translocation of proteins for degradation)5Serine/threonine-protein kinase 1OSR1Phosphorylates PAK1 (interacts with cortactin)6Ecotropic viral integration site protein 2BEVI2BInvolved in melanocyte and keratinocyte differentiation6Coiled-coil domain containing 111CCDC111Involved in DNA integrity and genome stability7Serine hydrolase-like 1SERHLPeroxisomal protein7Growth factor-induced 1GFI1Transcriptional repressor; involved in hematopoesis and oncogenesis Citation Format: Sean J. Leith, Susan E. Kuruvilla, Jason Moffat, Ann F. Chambers, Eva A. Turley, Joseph L. Chin, Hon S. Leong. A genome-wide shRNA screen for suppressors of prostate cancer cell invasion. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3432. doi:10.1158/1538-7445.AM2014-3432

Immunohistochemical expression of heat shock proteins, p63 and androgen receptor in benign prostatic hyperplasia and prostatic carcinoma in the dog.

This study compared heat shock proteins Hsp60, Hsp72 and Hsp73, along with p63 and androgen receptor (AR) immunoexpression between 16 cases of benign prostatic hyperplasia (BPH) and 11 prostatic carcinomas (PCa) in dogs. The proportion of Hsp60-positive cells was higher in PCa compared with BPH (P = 0.033), whereas the frequency and intensity of Hsp73 immunostaining did not differ significantly between the two groups. Hsp72-immunostained nuclei formed a discontinuous layer along the basement membrane in BPH, whereas cells in this layer in PCa were negative or weakly positive. Hsp72 nuclear score showed significant positive associations with both p63 (P = 0.016) and AR (P = 0.009) scores. Double immunofluorescence revealed Hsp72-p63 and Hsp72-AR co-expressions in basal cell nuclei. Aberrant cytoplasmic p63 immunolabelling was observed in 3 of 11 PCa cases. These results suggest a role of the combined expression of Hsp72, p63 and AR in basal epithelial cells in canine BPH and PCa.

Lycopene induce apoptosis in human prostate cells and alters the expression of Bax and Bcl-2 genes

Prostate cancer is the most common malignancy in men and the second leading cause of cancer-related mortality in men of the Western world. Lycopene has received attention due to its potential to prevent cancer. Alterations of the balance between proliferation and apoptosis are associated with cancer. In the present study, we evaluated the influence of lycopene on cell viability and apoptosis of human prostate cancer (PCa) cells and benign prostate hyperplasia (BPH) cells. We have studied prostate cells treated with lycopene for 96 h. Using MTT assay, we observed a decrease of viable PCa cells after treatment with lycopene. Flow cytometer analysis revealed that lycopene promoted up to two-fold increase of apoptotic cells in PCa cells when compared to the control group. Using real time PCR assay, we found that lycopene promoted an up-regulation of Bax gene and a downregulation of Bcl-2 gene in PCa cells. Less expressive effects were observed in BPH cells. Taken together, the present study supports the proposal that lycopene may have a protective effect on prostate cancer.

Voltage-gated sodium channels were differentially expressed in human normal prostate, benign prostatic hyperplasia and prostate cancer cells.

Voltage-gated sodium channels (VGSCs) are expressed not only in excitable cells but also in numerous metastatic cells, particularly in certain types of cancer cells. In some types of cancer, including prostate cancer, the expression of VGSCs is associated with cancer migration, invasion and metastasis in vivo. However, the detailed expression profiles of VGSC α subunits in normal human prostate, in prostatic hyperplasia and prostatic cancer remain controversial. In the present study, quantitative polymerase chain reaction was used to systematically detect all subtypes of VGSC α subunits in normal human prostate, benign prostatic hyperplasia (BPH) and prostate cancer cells. The expression profile of VGSC α subunits was observed to differ between these cell types. Nav1.5 was the major isoform expressed in normal human prostate tissue, while Nav1.5 and Nav1.2 were the predominant isoforms in BPH tissue. However, in PC-3 and LNCaP cells, two typical prostate cancer cell lines, Nav1.6 and Nav1.7 were abundantly expressed. By comparing the relative expression levels of Nav1.5, Nav1.6 and Nav1.7 in these cells, the mRNA levels of Nav1.6 and Nav1.7 were identified to be 6- to 27-fold higher in PC-3 and LNCaP cells than in either normal or BPH samples (P<0.05); however, Nav1.5 mRNA levels were relatively lower compared with those of Nav1.6 or Nav1.7 in all cells analyzed. To confirm whether Nav1.6 and Nav1.7 expression in cancer cells was functional, a patch-clamp technique was used to record whole-cell currents. A tetrodotoxin-sensitive sodium current was successfully recorded in PC-3 cells, but not in LNCaP cells. It was concluded that although all types of VGSC α subunits exhibited low expression levels in normal prostate and BPH cells, both Nav1.6 and Nav1.7 were significantly upregulated in the prostate cancer cell lines, suggesting these subtypes may be potential diagnostic markers and therapeutic targets for certain types of prostate cancer in humans.

Evidence of TGF-β1 mediated epithelial-mesenchymal transition in immortalized benign prostatic hyperplasia cells

Expression of epithelial-mesenchymal transition (EMT) markers has been detected clinically in benign prostatic hyperplasia (BPH) tissues. To understand the molecular basis, we investigated the role of stromal microenvironment in the progression of EMT in BPH cells. First, we used cell culture supernatant from normal prostate stromal WPMY-1 cells to provide supernatant-conditioned medium (WSCM) to culture the BPH-1 cell line. Then, the morphological changes and migratory capacity were detected in BPH-1 cells. The expression of EMT markers was examined in BPH-1 cells by Western blot and immunofluorescent analysis. Finally, to investigate the role of transforming growth factor beta 1 (TGF-β1) in this process, the WSCM-cultured cells were treated with monoclonal antibody against TGF-β1 to study its effect on EMT. We found that the morphology of BPH-1 cells changed to a spindle-like shape after cultured in WSCM, and the levels of E-cadherin and cytokeratin 5/8 (CK5/8) were significantly lower than the cells cultured in ordinary medium. These BPH-1 cells were also tested positive for mesenchymal markers vimentin and a-smooth muscle actin (SMA) as well as Snail. We also found WSCM can increase the migratory capacity of BPH-1 cells. In addition, when they were treated with anti-TGF-β1, upregulation of E-cadherin and CK5/8 levels was observed but no expression of vimentin, alpha-SMA or Snail was detected. Furthermore, phosphorylated-Smad3 expression in WSCM-cultured BPH-1 cells was also suppressed by anti-TGF-β1 treatment. Our results demonstrated that stromal cell supernatant was able to induce EMT in BPH-1 cells, possibly through secreting TGF-β1 to activate Smad signaling. Our results suggest novel molecular targets for clinical treatment of BPH by modification of stromal microenvironment through inhibiting TGF-β1/Smad expression.