Normal Transition Zone Research Articles

MP17-14 DEPLETION OF PERIPHERAL SEROTONIN SYNTHESIS INDUCES BENIGN PROSTATIC GROWTH IN MICE: MORE EVIDENCE FOR THE NEW “NEUROENDOCRINE THEORY” IN BPH ETIOLOGY

You have accessJournal of UrologyBenign Prostatic Hyperplasia: Basic Research & Pathophysiology1 Apr 2017MP17-14 DEPLETION OF PERIPHERAL SEROTONIN SYNTHESIS INDUCES BENIGN PROSTATIC GROWTH IN MICE: MORE EVIDENCE FOR THE NEW “NEUROENDOCRINE THEORY” IN BPH ETIOLOGY Paulo Mota, Emanuel Carvalho-Dias, Alice Miranda, Olga Martinho, Cristina Nogueira-Silva, Natalia Alenina, Michael Bader, Riccardo Autorino, Estevão Lima, and Jorge Correia-Pinto Paulo MotaPaulo Mota More articles by this author , Emanuel Carvalho-DiasEmanuel Carvalho-Dias More articles by this author , Alice MirandaAlice Miranda More articles by this author , Olga MartinhoOlga Martinho More articles by this author , Cristina Nogueira-SilvaCristina Nogueira-Silva More articles by this author , Natalia AleninaNatalia Alenina More articles by this author , Michael BaderMichael Bader More articles by this author , Riccardo AutorinoRiccardo Autorino More articles by this author , Estevão LimaEstevão Lima More articles by this author , and Jorge Correia-PintoJorge Correia-Pinto More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2017.02.603AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Ageing and testosterone cause almost inexorably benign prostatic hyperplasia (BPH) in Human males, however the etiology of BPH is unknown. Serotonin (5-HT) is produced by neuroendocrine prostatic cells and is present in high concentration in normal prostatic transition zone. In BPH neuroendocrine cells and 5-HT are significantly decreased comparatively to normal prostatic transition zone. Previously, we have demonstrated in several in vitro models that 5-HT inhibitis non-malignant prostatic growth through androgen receptor down-regulation and we suggested a new Neuroendocrine Theory for BPH etiology. Here, we investigated in vivo the effects of peripheral inhibition of 5-HT synthesis on mice prostate gland. METHODS peripheral 5-HT synthesis is critical dependent of the presence of TPH1, so we used transgenic mice depleted from TPH1 (Tph1-/-) to study the in vivo effect of peripheral serotonin depletion. Male wild-type and Tph1-/- mice were sacrificed at different time points: 7, 12, 16 and 20 weeks-old. For pharmacological studies, wild-type and Tph1-/- mice with 19 week-old were treated with daily intraperitoneal injections of 0,9% saline or 5-HT (100 mg/Kg) during 10 consecutive days.Prostate gland mass was determined and proceeded for histology, western blotting, immunofluorescence and qRT-PCR of AR expression. RESULTS We showed that Tph1 knockout mice depleted from peripheral 5-HT have significant higher prostate mass comparatively to wild-type (p < 0,001) and 5-HT treatment of Tph1 knockout mice restores prostate mass to levels of wild-type (p < 0,001) (figure A and B). We demonstrated also that the benign prostatic growth in Tph1 knockout mice is associated with up-regulation of AR and 5-HT treatment restores de expression of AR (p<0,05) (Figure C and D). CONCLUSIONS In vivo, 5-HT depletion induces benign prostatic growth in mice trough AR up-regulation. 5-HT depletion in transition zone of aging human male could be the etiologic factor for BPH etiology. © 2017FiguresReferencesRelatedDetails Volume 197Issue 4SApril 2017Page: e216-e217 Advertisement Copyright & Permissions© 2017MetricsAuthor Information Paulo Mota More articles by this author Emanuel Carvalho-Dias More articles by this author Alice Miranda More articles by this author Olga Martinho More articles by this author Cristina Nogueira-Silva More articles by this author Natalia Alenina More articles by this author Michael Bader More articles by this author Riccardo Autorino More articles by this author Estevão Lima More articles by this author Jorge Correia-Pinto More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Patient-specific pharmacokinetic parameter estimation on dynamic contrast-enhanced MRI of prostate: Preliminary evaluation of a novel AIF-free estimation method.

To develop and evaluate a prostate-based method (PBM) for estimating pharmacokinetic parameters on dynamic contrast-enhanced (DCE) magnetic resonance imaging (MRI) by leveraging inherent differences in pharmacokinetic characteristics between the peripheral zone (PZ) and transition zone (TZ). This retrospective study, approved by the Institutional Review Board, included 40 patients who underwent a multiparametric 3T MRI examination and subsequent radical prostatectomy. A two-step PBM for estimating pharmacokinetic parameters exploited the inherent differences in pharmacokinetic characteristics associated with the TZ and PZ. First, the reference region model was implemented to estimate ratios of Ktrans between normal TZ and PZ. Subsequently, the reference region model was leveraged again to estimate values for Ktrans and ve for every prostate voxel. The parameters of PBM were compared with those estimated using an arterial input function (AIF) derived from the femoral arteries. The ability of the parameters to differentiate prostate cancer (PCa) from benign tissue was evaluated on a voxel and lesion level. Additionally, the effect of temporal downsampling of the DCE MRI data was assessed. Significant differences (P < 0.05) in PBM Ktrans between PCa lesions and benign tissue were found in 26/27 patients with TZ lesions and in 33/38 patients with PZ lesions; significant differences in AIF-based Ktrans occurred in 26/27 and 30/38 patients, respectively. The 75th and 100th percentiles of Ktrans and ve estimated using PBM positively correlated with lesion size (P < 0.05). Pharmacokinetic parameters estimated via PBM outperformed AIF-based parameters in PCa detection. J. Magn. Reson. Imaging 2016;44:1405-1414.

Sand body types of clastic shore deposits with delta backgrounds and their control over reservoir quality

The “Donghe sandstones” of the Tarim Basin are not simple shore deposits. They show distinct sedimentary characteristics and different types of sand bodies that are controlled by modes developed in different areas. This study provides an example from the Bachu Formation, which consists of the “Donghe sandstones” deposits located in the Donghetang Oilfield in the Tarim Basin, China. Based on analyses, cores, logs, and seismic data, a new depositional system is proposed for the Donghetang section as a wave-dominated shore deposit with delta backgrounds. The sedimentary characteristics of the transitional shoreland, called “Three types of bar, two types of beach and two types of sands in the transitional zone,” were summarized by the further classification of the shoreface bars, beaches, and transitional sands on the basis of their origin types and varying degrees of wave reworking. The classifications are bars from the reworked distributary channels and mouth bars, normal bars, beaches from the delta sands, normal beaches, sands from reworked delta sheet sands, and normal transitional zone sands. Meanwhile, the Donghetang section in the study area is interpreted as wave-dominated shore deposits with delta backgrounds rather than normal shore deposits, which has significant implications for the distribution of the favorable reservoirs. The shoreface bars, formed from reworked distributary channels and mouth bars, and the shoreface beaches, formed from delta sands, have better connectivity and constitute the additional important reservoirs.

Rapid quantitative T2 mapping of the prostate using three-dimensional dual echo steady state MRI at 3T.

To develop and evaluate a rapid three-dimensional (3D) quantitative T2 mapping method for prostate cancer imaging using dual echo steady state (DESS) MRI at 3T. In simulations, DESS-T2 mapping in the presence of T1 and B1+ variations was evaluated. In a phantom and in healthy volunteers (n = 4), 3D DESS-T2 mapping was compared with a two-dimensional turbo spin echo (TSE) approach. In volunteers and a pilot patient study (n = 29), quantitative T2 in normal prostate anatomical zones and in suspected cancerous lesions was evaluated. The simulated bias for DESS-T2 was < 2% (5%) for typically observed T1 ( B1+) variations. In phantoms and in vivo, high correlation of DESS-T2 and TSE-T2 (r2 = 0.98 and 0.88, P < 0.001) was found. DESS-T2 in the normal peripheral zone and transition zone was 115 ± 26 ms and 64 ± 7 ms, respectively, in healthy volunteers and 129 ± 39 ms and 83 ± 12 ms, respectively, in patients. In suspected cancerous lesions, DESS-T2 was 72 ± 14 ms, which was significantly decreased from the normal peripheral zone (P < 0.001) but not from the transition zone. Rapid 3D T2 mapping in the entire prostate can be performed in 1 min using DESS MRI. Magn Reson Med 76:1720-1729, 2016. © 2016 International Society for Magnetic Resonance in Medicine.

Oxidative stress promotes benign prostatic hyperplasia.

Benign prostatic hyperplasia (BPH) is characterized by increased tissue mass in the transition zone of the prostate, which leads to obstruction of urine outflow and significant morbidity in the majority of older men. Plasma markers of oxidative stress are increased in men with BPH but it is unclear whether oxidative stress and/or oxidative DNA damage are causal in the pathogenesis of BPH. Levels of 8-OH deoxyguanosine (8-OH dG), a marker of oxidative stress, were measured in prostate tissues from normal transition zone and BPH by ELISA. 8-OH dG was also detected in tissues by immunohistochemistry and staining quantitated by image analysis. Nox4 promotes the formation of reactive oxygen species. We therefore created and characterized transgenic mice with prostate specific expression of Nox4 under the control of the prostate specific ARR2PB promoter. Human BPH tissues contained significantly higher levels of 8-OH dG than control transition zone tissues and the levels of 8-OH dG were correlated with prostate weight. Cells with 8-OH dG staining were predominantly in the epithelium and were present in a patchy distribution. The total fraction of epithelial staining with 8-OH dG was significantly increased in BPH tissues by image analysis. The ARR2PB-Nox4 mice had increased oxidative DNA damage in the prostate, increased prostate weight, increased epithelial proliferation, and histological changes including epithelial proliferation, stromal thickening, and fibrosis when compared to wild type controls. Oxidative stress and oxidative DNA damage are important in the pathogenesis of BPH.

Nucleoporins but not septins at the transition zone of cilia in Paramecium and Tetrahymena

The transition zone (TZ) of locomotory cilia participates in docking young basal bodies to the surface, stabilizes the basal body-membrane connection, forms a diffusion barrier and modulates the intraciliary transport. In vertebrate ciliated cells, nucleoporins and septins were suggested to be present in the TZ of locomotory and primary cilia. Here we present data suggesting the presence of nucleoporins, at both nuclear pores and ciliary TZ, in Tetrahymena and Paramecium. Using the monoclonal 414, specific to FG-containing nucleoporins, and anti-tubulin antibodies, we detected labeling at the distal part of basal bodies and as a spotted pattern around nuclei. In previous studies on Tetrahymena, GFP-tagged septins (Sep1, Sep2 and Sep3) localized to mitochondrial/ER compartments but neither to basal bodies, cilia nor nuclei. Tetrahymena cells with knocked out septins display disrupted nuclear (Mac and Mic) membranes, abnormal mitochondria, but unaffected ciliary TZ. Similarly, in Paramecium, silencing of the SEP2 gene revealed affected nuclear membranes and mitochondria but normal TZ, and the mAb 414 and DAPI showed a normal labeling of the ciliary base, but patches of missing nuclear pores in the macronucleus. Our studies support the conclusion that some nucleoporins are present in both nuclear pores and ciliary TZ. In addition, the nuclear pore proteins seem to interact with septins, yet the lack of effect of septin silencing on the localization of the mAb 414 in the ciliary TZ suggests that nucleoporins may have different binding partners in nuclear pores and ciliary TZ.

Differentially Expressed Androgen-Regulated Genes in Androgen-Sensitive Tissues Reveal Potential Biomarkers of Early Prostate Cancer

BackgroundSeveral data favor androgen receptor implication in prostate cancer initiation through the induction of several gene activation programs. The aim of the study is to identify potential biomarkers for early diagnosis of prostate cancer (PCa) among androgen-regulated genes (ARG) and to evaluate comparative expression of these genes in normal prostate and normal prostate-related androgen-sensitive tissues that do not (or rarely) give rise to cancer.MethodsARG were selected in non-neoplastic adult human prostatic epithelial RWPE-1 cells stably expressing an exogenous human androgen receptor, using RNA-microarrays and validation by qRT-PCR. Expression of 48 preselected genes was quantified in tissue samples (seminal vesicles, prostate transitional zones and prostate cancers, benign prostatic hypertrophy obtained from surgical specimens) using TaqMan® low-density arrays. The diagnostic performances of these potential biomarkers were compared to that of genes known to be associated with PCa (i.e. PCA3 and DLX1).Results and DiscussionBy crossing expression studies in 26 matched PCa and normal prostate transitional zone samples, and 35 matched seminal vesicle and PCa samples, 14 genes were identified. Similarly, 9 genes were overexpressed in 15 benign prostatic hypertrophy samples, as compared to PCa samples. Overall, we selected 8 genes of interest to evaluate their diagnostic performances in comparison with that of PCA3 and DLX1. Among them, 3 genes: CRYAB, KCNMA1 and SDPR, were overexpressed in all 3 reference non-cancerous tissues. The areas under ROC curves of these genes reached those of PCA3 (0.91) and DLX1 (0.94).ConclusionsWe identified ARG with reduced expression in PCa and with significant diagnostic values for discriminating between cancerous and non-cancerous prostatic tissues, similar that of PCA3. Given their expression pattern, they could be considered as potentially protective against prostate cancer. Moreover, they could be complementary to known genes overexpressed in PCa and included along with them in multiplex diagnostic tools.

Human intraretinal myelination: Axon diameters and axon/myelin thickness ratios

Purpose:Human intraretinal myelination of ganglion cell axons occurs in about 1% of the population. We examined myelin thickness and axon diameter in human retinal specimens containing myelinated retinal ganglion cell axons.Materials and Methods:Two eyes containing myelinated patches were prepared for electron microscopy. Two areas were examined in one retina and five in the second retina. Measurements were compared to normal retinal and optic nerve samples and the rabbit retina, which normally contains myelinated axons. Measurements were made using a graphics tablet.Results:Mean axon diameter of myelinated axons at all locations were significantly larger than unmyelinated axons (P ≤ 0.01). Myelinated axons within the patches were significantly larger than axons within the optic nerve (P < 0.01). The relationship between axon diameter/fiber diameter (the G-ratio) seen in the retinal sites differed from that in the nerve. G-ratios were higher and myelin thickness was positively correlated to axon diameter (P < 0.01) in the retina but negatively correlated to axon diameter in the nerve (P < 0.001).Conclusion:Intraretinally myelinated axons are larger than non-myelinated axons from the same population and suggests that glial cells can induce diameter changes in retinal axons that are not normally myelinated. This effect is more dramatic on intraretinal axons compared with the normal transition zone as axons enter the optic nerve and these changes are abnormal. Whether intraretinal myelin alters axonal conduction velocity or blocks axonal conduction remains to be clarified and these issues may have different clinical outcomes.

Microvascularity in transition zone prostate tumors resembles normal prostatic tissue

The objective of this study was comparison of characteristics of the microvasculature in transition zone tumor (TZT) and benign nodular hyperplasia (BPH) with normal prostatic transition zone (NTZ), applying accurate and objective quantification based on digital image analysis. Results of this study may increase understanding of prostate dynamic contrast enhanced (DCE) MRI analysis. Radical prostatectomy specimens of 28 patients containing TZT ranging from pT2-pT4 were used. In 11 patients a concomitant peripheral zone tumor (PZT) was present. Microvessels were visualized by CD31 immunohistochemistry. Specimens were scanned using a computer-controlled microscope with automatic recognition of microvessels. Pseudocolor maps were produced displaying microvessel density, perimeter, and area of an entire prostate transection. Mean, 75th percentile (p75) and coefficient of variation (CV) were calculated automatically in manually indicated areas of the tumor and corresponding contralateral normal tissue, and BPH. Large variability was seen in TZT microvascular parameters, indicating presence of patients having both hypo and hypervascularized tumors compared to NTZ. In contrast, areas of BPH showed a more consistent increase in vascular parameters, with decreased CV. Analysis of PZT confirmed results of our previous study, with mean and p75 of all vascular parameters being significantly increased and a decrease in CV. No correlation was found for clinicopathological parameters and microvascular parameters. Microvasculature of transition zone tumor showed increased heterogeneity compared to BPH and peripheral zone tumors, possibly explaining the difficulty of TZT detection on DCE-MRI.

Seismic signatures of an altered crust and a normal transition zone structure beneath the Godavari rift

The Godavari rift valley in the south Indian shield, located over a paleosuture between the Dharwar and Singhbhum cratons is a very less studied region in terms of its deep seismic structure. In the present study a total of 1559 receiver functions from 777 earthquakes registered by a 10 station network are utilized to investigate crust and mantle structure. Broadly, the crustal architecture underneath the rift appears very complex with the presence of intracrustal layers and a weak Moho compared to a sharp Moho and a simple crust outside it. Detailed modelling of receiver functions at 6 stations using the Nearest Neighbourhood Algorithm approach reveals a 40km thick crust within the rift compared to a 33km thick crust in the adjacent Dharwar craton. Also, the lower crustal character is found to be distinctly different, with a faster lower crust beneath the rift. A weak Moho beneath the rift may be due to magmatic underplating. The transition zone (660–410km) structure inferred from the common conversion point stacks of receiver functions can be interpreted in terms of the absence of (a) thermal anomalies within and above the mantle transition zone (MTZ) (b) thin lithosphere or high temperatures in the uppermost mantle. Magmatic underplating within the crust and a normal MTZ argue in favour of a rift formed due to lithospheric stretching without the influence of a mantle plume.

FOXA1 Promotes Tumor Progression in Prostate Cancer and Represents a Novel Hallmark of Castration-Resistant Prostate Cancer

Forkhead box protein A1 (FOXA1) modulates the transactivation of steroid hormone receptors and thus may influence tumor growth and hormone responsiveness in prostate cancer. We therefore investigated the correlation of FOXA1 expression with clinical parameters, prostate-specific antigen (PSA) relapse-free survival, and hormone receptor expression in a large cohort of prostate cancer patients at different disease stages. FOXA1 expression did not differ significantly between benign glands from the peripheral zone and primary peripheral zone prostate carcinomas. However, FOXA1 was overexpressed in metastases and particularly in castration-resistant cases, but was expressed at lower levels in both normal and neoplastic transitional zone tissues. FOXA1 levels correlated with higher pT stages and Gleason scores, as well as with androgen (AR) and estrogen receptor expression. Moreover, FOXA1 overexpression was associated with faster biochemical disease progression, which was pronounced in patients with low AR levels. Finally, siRNA-based knockdown of FOXA1 induced decreased cell proliferation and migration. Moreover, in vitro tumorigenicity was inducible by ARs only in the presence of FOXA1, substantiating a functional cooperation between FOXA1 and AR. In conclusion, FOXA1 expression is associated with tumor progression, dedifferentiation of prostate cancer cells, and poorer prognosis, as well as with cellular proliferation and migration and with AR signaling. These findings suggest FOXA1 overexpression as a novel mechanism inducing castration resistance in prostate cancer.

P16/CDKN2A and Ki-67 Enhance the Detection of Anal Intraepithelial Neoplasia and Condyloma and Correlate With Human Papillomavirus Detection by Polymerase Chain Reaction

The classification of anal intraepithelial neoplasia (AIN) in mucosal biopsies is subject to considerable interobserver variability. Previous studies have shown that Ki-67 and p16/CDKN2A immunostains aid detection of dysplasia in biopsy samples from the uterine cervix. The aim of this study was to determine whether Ki-67 and p16/CDKN2A immunolabeling enhanced diagnostic accuracy in the assessment of anal mucosal biopsies from patients with suspected human papillomavirus (HPV) infection. The study consisted of 75 cases that were originally interpreted to represent normal anal transitional zone (n=15), fibroepithelial polyp (n=10), condyloma accuminatum (n=10), low-grade AIN (AIN1, n=20), and high-grade AIN (n=20), including 10 cases each of AIN2 and AIN3. The histologic features of all cases were re-reviewed and categorized based upon consensus agreement, which resulted in reclassification of 16 cases. Thus, the final study group consisted of 17 samples of normal anal transition zone, 14 fibroepithelial polyps, 6 condylomata accuminata, and 38 cases of AIN (11 AIN1, 16 AIN2, and 11 AIN3). Each case was tested for the presence of HPV DNA by a SPF10 polymerase chain reaction and LiPA25 genotyping assay and immunostained for Ki-67 and p16/CDKN2A. A positive Ki-67 result was defined as the presence of a cluster of at least 2 strongly stained epithelial nuclei in the upper two-thirds of the epithelial thickness. A positive result for p16/CDKN2A was defined as diffuse moderate-to-strong cytoplasmic and nuclear staining. None of the anal transition zone samples or fibroepithelial polyps showed Ki-67 or p16/CDKN2A staining. All condylomata and samples of AIN contained HPV DNA and showed positive Ki-67 labeling. All cases of high-grade AIN showed positive p16/CDKN2A staining. We conclude that Ki-67 labeling detects anal HPV-related changes with a high degree of sensitivity and specificity, whereas increased p16/CDKN2A staining is strongly associated with high-grade squamous neoplasia. These results indicate that a combination of these markers may aid interpretation of anal mucosal biopsy samples.

Spectro-IRM de prostate avec antennes externes : faisabilité en pratique clinique ?

To determine the clinical feasibility of MR spectroscopy (MRS) of prostate cancer using external multi-channel surface coils at 1.5T. Materials and methods. Retrospective study of 31 patients with prostate cancer who underwent MRS as part of the staging work-up prior to radical prostatectomy. The ratio of the three main metabolites ([choline + creatine]/citrate) were measured along with spectral analysis of different regions of interest (ROI) placed in areas of normal tissue and cancer using the surgical specimen as the standard of reference. One hundred and eighty-three voxels were analyzed. Qualitative visual analysis identified pathological spectra in 88.5% of cancer ROI, 11.7% of normal transitional zone ROI and 1.6% of normal peripheral zone ROI. The ratios of normal prostate tissue were significantly more elevated (p<0.0001) in the transition zone (0.41 +/- 0.24) than in the peripheral zone (0.22 +/- 0.36). Tumor containing voxels had significantly higher ratios (2.84 +/- 2.74) than normal tissue containing voxels of the transition zone (p<0.0001) and peripheral zone (p<0.0001). Prostate MRS using external surface coils appears routinely feasible at 1.5T, even though it presents some limitations.

Gene expression of forkhead transcription factors in the normal and diseased human prostate

OBJECTIVE To assess the expression of forkhead transcription factors (FOX) in normal prostate and prostate diseases, as since the first FOX was identified, its family members have been implicated in a variety of cellular processes, including embryonic development and disease. MATERIAL AND METHODS We analysed a set of 12 different FOX genes by quantitative reverse transcription-polymerase chain reaction in prostate zones, prostate cancer, lymph node metastases, benign prostatic hyperplasia (BPH), xenografts and several prostate cell lines. RESULTS There were striking differences among the expression of various FOX family members; most prominent were the high expression of FOXF1 and FOXF2 in the normal prostate transition zone and BPH, and their decreased expression in prostate cancer. Interestingly, although the FOXF genes are stroma-specific, some of the androgen-independent prostate cancer xenografts uniquely express these two genes. FOXD1 and FOXD2 were more highly expressed in prostate cancer and lymph node metastases. FOXA1 and FOXC1 have an opposite expression pattern for androgen-dependent growth of prostate cancer cell lines and xenografts. CONCLUSIONS Various members of the FOX family are differentially expressed in the zones of the normal prostate and in benign and malignant outgrowths. The expression profiles of FOXF1 and FOXF2 suggest a role in epithelial to mesenchymal transition, while FOXA1 and FOXC1 expression is linked to androgen-associated growth status of cancer.

Tumor formation of prostate cancer cells influenced by stromal cells from the transitional or peripheral zones of the normal prostate

This study was designed to investigate the different involvements of prostatic stromal cells from the normal transitional zone (TZ) or peripheral zone (PZ) in the carcinogenesis of prostate cancer (PCa) epithelial cells (PC-3) in vitro and in vivo co-culture models. Ultra-structures and gene expression profiles of primary cultures of human prostatic stromal cells from the normal TZ or PZ were analyzed by electron microscopy and microarray analysis. In vitro and in vivo co-culture models composed of normal TZ or PZ stromal cells and human PCa PC-3 cells were established. We assessed tumor growth and weight in the in vivo nude mice model. There are morphological and ultra-structural differences in stromal cells from TZ and PZ of the normal prostate. In all, 514 differentially expressed genes were selected by microarray analysis; 483 genes were more highly expressed in stromal cells from TZ and 31 were more highly expressed in those from PZ. Co-culture with PZ stromal cells and transforming growth factor-beta1 (TGF-beta1) increased the tumor growth of PC-3 cells in vitro and in vivo, as well as Bcl-2 expression. On the other hand, stromal cells of TZ suppressed PC-3 cell tumor growth in the mouse model. We conclude that ultra-structures and gene expression differ between the stromal cells from TZ or PZ of the normal prostate, and stroma-epithelium interactions from TZ or PZ might be responsible for the distinct zonal localization of prostate tumor formation.

Oesophageal peristaltic transition zone defects: real but few and far between

This study analysed the association between oesophageal transition zone (TZ) defects [characterized by a delay and/or spatial gap between the terminus of the proximal oesophageal (striated muscle) contraction and the initiation of the distal oesophageal (smooth muscle) contraction] and dysphagia in a large patient cohort. Four hundred consecutive patients (178 with dysphagia) and 75 controls were studied with 36-channel high-resolution manometry (HRM). The resultant pressure topography plots were first analysed for impaired oesophagogastric junction (OGJ) relaxation, distal segment contractile abnormalities, and proximal contractile abnormalities using normal values from the 75 controls. If these aspects of oesophageal motility were deemed normal, the TZ was characterized by length and duration between the proximal and distal contractions using a 20 mmHg isobaric contour to establish the segment boundaries. Patients were then classified according to whether or not they exhibited TZ defects (spatial separation or delay) and the occurrence of unexplained dysphagia. Of the 400 patients, 267 were suitable for TZ analysis and of these 55 had a spatial or temporal TZ measurement exceeding the 95th percentile of the controls (2 cm, 1 s). Exactly 34.6% of the patients (n = 19) with spatial and/or temporal TZ defects had unexplained dysphagia, which was significantly more than seen with normal TZ dimensions (19.8%). Although far less common than distal peristaltic or OGJ abnormailites, TZ defects may be related to dysphagia in a minority of patients (<4% in this series) and should be considered a distinct oesophageal motility disorder.

Elevated Epithelial Expression of Interleukin-8 Correlates with Myofibroblast Reactive Stroma in Benign Prostatic Hyperplasia

Numerous inflammatory diseases display elevated interleukin (IL)-8, and most are associated with a reactive stroma. IL-8 expression is also elevated in benign prostatic hyperplasia (BPH), yet little is known about reactive stroma in BPH. Whether a reactive stroma response exists in BPH, whether this correlates with elevated IL-8, and whether IL-8 can induce a reactive stroma phenotype have not been determined. This study was designed to specifically address these issues. Normal prostate transition zone tissue and BPH specimens, as identified by the Baylor College of Medicine pathology department, were examined by quantitative immunohistochemistry to correlate IL-8, smooth muscle alpha-actin, vimentin, calponin, and tenascin-C. In addition, human prostate stromal cell cultures were used to evaluate the effect of IL-8 on the expression of reactive stroma biomarkers. BPH nodules exhibited elevated epithelial IL-8 immunoreactivity, and this correlated with elevated smooth muscle alpha-actin, reduced calponin, and altered deposition of tenascin-C, relative to the normal prostate transition zone tissue (P <0.05). Multiple vimentin-positive prostate stromal fibroblast cultures were induced by IL-8 to also co-express smooth muscle alpha-actin and tenascin-C, typical of a reactive stroma myofibroblast phenotype. These data show that BPH reactive stroma is fundamentally different from normal prostate fibromuscular stroma and is typified by the emergence of a reactive stroma myofibroblast phenotype. This reactive stroma pattern correlated spatially with IL-8 elevation in adjacent epithelium. Additionally, IL-8 induced expression of myofibroblast markers in human prostate fibroblasts in vitro. These results suggest that IL-8 acts as a regulator of BPH reactive stroma and is therefore a potential therapeutic target.

Distinctive gene expression of prostatic stromal cells cultured from diseased versus normal tissues

To obtain a comprehensive view of the transcriptional programs in prostatic stromal cells of different histological/pathological origin, we profiled 18 adult human stromal cell cultures from normal transition zone (TZ), normal peripheral zone (PZ), benign prostatic hyperplasia (BPH), and prostate cancer (CA) using cDNA microarrays. A hierarchical clustering analysis of 714 named unique genes whose expression varied at least threefold from the overall mean abundance in at least three samples in all 18 samples demonstrated that cells of different origin displayed distinct gene expression profiles. Many of the differentially expressed genes are involved in biological processes known to be important in the development of prostatic diseases including cell proliferation and apoptosis, cell adhesion, and immune response. Significance Analysis of Microarrays (SAM) analysis identified genes that showed differential expression with statistical significance including 24 genes between cells from TZ versus BPH, 34 between BPH versus CA, and 101 between PZ versus CA. S100A4 and SULF1, the most up- and downregulated genes in BPH versus TZ, respectively, showed expression at the protein level consistent with microarray analysis. In addition, sulfatase assay showed that BPH cells have lower SULF1 activity compared to TZ cells. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis confirmed differential expression of ENPP2/autotoxin and six other genes between PZ versus CA, as well as differential expression of six genes between BPH versus CA. Our results support the hypothesis that prostatic stromal cells of different origin have unique transcriptional programs and point towards genes involved in actions of stromal cells in BPH and CA.

Expression of different genes in transitional zone and peripheral zone of human normal prostate

To detect the expression of different genes in the transitional zone and peripheral zone of human normal prostate. Seventeen specimens of normal prostate gland were obtained from dead kidney donors and patients undergoing total cystectomy. The transitional zone and peripheral zone were isolated. Twenty specimens of benign prostate hypertrophy (BPH) were obtained from patients undergoing prostatic enucleation. The periurethral significantly hypertrophic nodules were isolated. BioDoor Chip-40S containing 6700 clones of targeting gene cDNA was used to detect the gene expression. The peripheral zone RNA samples were labeled with cy3, and the transitional zone RNA samples labeled with Cy5. The genes with the ratio average (RA) < 0.5 or > 2 were identified as differentially expressed genes, and those with a RA < 0.2 or > 4 were identified as the most significantly differentially expressed genes. RT-PCR was used to investigate semi-quantitatively the expression of epidermal growth factor (EGF) mRNA in the 17 specimens of transitional zone and peripheral zone from the 17 normal prostate glands and the 20 specimens of periurethral zone from BPH. A total of 640 genes differentially expressed in transitional zone and peripheral zone were screened out (comprising 269 unknown genes and 371 known genes), of which 294 genes were expressed in the transitional zone and 346 in the peripheral zone. Fourteen genes were identified as markedly differentially expressed, among which the EGF gene was highly expressed in the central zone, and the OSF-1 (osteoblast stimulating factor-1) gene was highly expressed in the peripheral zone. RT-PCR proved that in comparison with the expression of the internal marker beta-actin gene, the expression levels of EGF in the normal transitional zone and in the periurethral zone from BPH were markedly higher than that in the peripheral zone. There are differently expressed genes between the peripheral zone and central zone of normal human prostate, which may be the background of different biological behavior between these zones. EGF may markedly contribute to the growth of central zone of normal prostate.

Analysis of microdissected prostate tissue with ProteinChip® arrays - a way to new insights into carcinogenesis and to diagnostic tools

Prostate carcinomas are one of the most common malignancies in western societies. The pathogenesis of this tumor is still poorly understood. These tumors present with two characteristic features: epithelial-mesenchymal interactions, which play a pivotal role for tumor development and most of clinically manifest cancers arise in prostate proper compared to a minority of tumors which develop in the transitional zone. Deciphering the epithelial-mesenchymal cross talk and identification of molecular pecularities of the sub-populations of cells in different zones can therefore help understanding carcinogenesis and development of new, non-invasive tools for the diagnosis and prognosis of prostate carcinomas which has remained a challenge until today. A ProteinChip array technology (SELDI = surface enhanced laser desorption ionization) has been developed recently by Ciphergen Biosystems enabling analysis and profiling of complex protein mixtures from a few cells. This study describes the analysis of approximately 500-1000 freshly obtained prostate cells by SELDI-TOF-MS (surface enhanced laser desorption ionization time-of-flight mass spectrometry). Pure cell populations of stroma, epithelium and tumor cells were selected by laser assisted microdissection. Multiple specific protein patterns were reproducibly detected in the range from 1.5 to 30 kDa in 28 sub-populations of 4 tumorous prostates and 1 control. A specific 4.3 kDa peak was increased in the prostate tumor stroma compared to normal prostate proper and transitional zone stroma and increased in prostate tumor glands compared to normal prostate proper and transitional zone glands. Coupling laser assisted microdissection with SELDI provides tremendous opportunities to identify cell and tumor specific proteins to understand molecular events underlying prostate carcinoma development. It underlines the vast potential of this technology to better understand pathogenesis and identify potential candidates for new specific biomarkers in general which could help to screen for and distinguish disease entities, i.e. between clinically significant and insignificant carcinomas of the prostate.