Increased risk of prostate cancer and benign prostatic hyperplasia associated with transforming growth factor-beta 1 gene polymorphism at codon10

Transforming growth factor-beta 1 (TGF-beta1) plays a significant role in regulating the proliferation and apoptosis of prostate epithelial and stromal cells. We explored the association between the T (Leu) to C (Pro) polymorphism at codon10 of the TGF-beta1 gene (TGFB1) and the risk of prostate cancer (PCa) or benign prostatic hyperplasia (BPH) in 351 PCa patients, 221 BPH patients and 303 male controls in Japan. There were significant differences in the CC versus TC + TT genotype distribution between PCa patients and male controls (P=0.008), and between BPH patients and male controls (P=0.041). Males with the TC or TT genotype had a 1.62-fold increased risk of PCa [95% confidence interval (95% CI)=1.14-2.30, P=0.007] and a 1.51-fold increased risk of BPH (95% CI=1.02-2.24, P=0.041) compared with those with the CC genotype, therefore suggesting the dominant effect of the TGFB1 T allele on development of PCa and BPH. There were no significant differences in the TGFB1 genotype distribution between different groups of tumor grades and stages in the PCa patients and no significant differences when PCa patients were stratified by the age of onset. The results suggest that the codon10 polymorphism in TGFB1 may have a significant influence on the development of PCa and BPH, therefore underscoring the importance of the TGF pathway in the development of these prostatic diseases. However, it appeared to have no impact on the disease status or age of onset of PCa.     

Real-time quantitative RT-PCR assessment of PIM-1 and hK2 mRNA expression in benign prostate hyperplasia and prostate cancer

To investigate the expressions of PIM-1 and hK2 mRNA in normal prostate, benign prostatic glandular hyperplasia (BPH), and prostate cancer (PCa), and to explore the association of PIM-1 and hK2 expressions with PCa progression. The samples were harvested from 37 patients with BPH, 23 patients with PCa, and three with normal prostate tissues. Total RNA was extracted from their prostate tissues and analyzed for PIM-1 and hK2 mRNA levels using SYBR green I-based quantitative real-time RT-PCR (QRT-PCR) assays and Southern blot analysis. The differences of gene expressions were calculated based on standard curve. Quantitative expressions of PIM-1 and hK2 mRNA in normal prostate, BPH, and PCa were 1.05 ± 0.04, 2.57 ± 0.74, 4.45 ± 0.63, and 1.02 ± 0.03, 2.264 ± 0.46, 5.905 ± 0.78, respectively. PIM-1 and hK2 were expressed higher in PCa than those in BPH and normal prostate tissues, the differences among which had statistic significance (P < 0.05). Our results support the hypothesis that PIM-1 and hK2 play a significant role in the growth of PCa and the detection of PIM-1 and hK2 mRNA expressions by QRT-PCR provided more reliable and helpful information on diagnosis, treatment, and prognosis of PCa. Hui-chan He and Xue-cheng Bi contributed equally to this article.     

加兰肽在前列腺癌组织中的表达及其意义

 目的　研究加兰肽（GLA）在前列腺癌组织中的表达及其临床意义。方法　采用免疫组织化学法检测GLA在50例前列腺增生、50例前列腺癌和30例发生骨转移的前列腺癌组织中的表达。结果　GLA在前列腺增生、前列腺癌及骨转移前列腺癌组织中阳性表达率分别为18 %（9／50）、68 %（34／50）和80 %（24／30）,前列腺癌及骨转移组阳性表达率高于前列腺增生组,组间比较差异有统计学意义（χ2值分别为25.5、29.74,均P＜0.01）,但骨转移组与前列腺癌组阳性表达率比较差异无统计学意义（χ2＝1.35,P＞0.05）。结论　GLA在前列腺癌组织中有较高表达,可作为良恶性前列腺疾病的鉴别指标,为前列腺癌的诊断及预测预后提供一定依据。     

Progression of prostate cancer: diagnostic and prognostic utility of prostate-specific antigen, alpha2-macroglobulin, and their complexes

We previously reported cases of advanced prostate cancer (PCa) in which serum alpha2-macroglobulin (alpha2M) levels were markedly decreased to less than approximately 50 mg/dl whereas serum prostate-specific antigen (PSA) levels were remarkably increased. These cases were not complicated with disseminated intravascular coagulation (DIC). In this study, we measured serum PSA and alpha2M in 108 patients with either benign prostatic hyperplasia (BPH) or PCa to elucidate the relationship between PSA, i.e. the serum protease derived from the prostatic tissue, and alpha2M, i.e. the protease inhibitor that was the most abundantly contained in serum. alpha2M was determined by ELISA, total PSA and PSA-alpha1-antichymotrypsin (PSA-ACT) by EIA, and free-PSA by RIA in 44 patients with untreated BPH and 64 patients with untreated PCa. The ready association of alpha2M and PSA was assessed using Western blotting to identify complexes of the two. Levels of total serum PSA correlated positively with those of PSA-ACT in PCa (r = 0.99, p < 0.001), and both levels increased with advancing stage of disease. In contrast, the serum-free PSA/total PSA ratio (free/total PSA) and alpha2M levels decreased as the disease progressed. However, only the free/total PSA ratio attained significant difference for localized cancer in stage T1,2 versus BPH (p < 0.05). In stage M1b PCa, in which serum PSA levels were very high, there was a negative correlation between the total PSA and alpha2M values (r = -0.57, p < 0.05). In addition, serum alpha2M levels tended to decrease with progression of PCa. Serum total PSA levels correlated tightly with serum PSA-ACT levels. It is suggested that PSA is usually complexed with ACT in the serum. Free/total PSA was useful for differential diagnosis between early cancer and BPH. Levels of serum alpha2M of less than 50 mg/dl in PCa patients may indicate a possibility of bone metastases.     

Use of multiple biomarkers for a molecular diagnosis of prostate cancer

The identification of biomarkers capable of providing a reliable molecular diagnostic test for prostate cancer (PCa) is highly desirable clinically. We describe here 4 biomarkers, UDP-N-Acetyl-alpha-D-galactosamine transferase (GalNAc-T3; not previously associated with PCa), PSMA, Hepsin and DD3/PCA3, which, in combination, distinguish prostate cancer from benign prostate hyperplasia (BPH). GalNAc-T3 was identified as overexpressed in PCa tissues by microarray analysis, confirmed by quantitative real-time PCR and shown immunohistochemically to be localised to prostate epithelial cells with higher expression in malignant cells. Real-time quantitative PCR analysis across 21 PCa and 34 BPH tissues showed 4.6-fold overexpression of GalNAc-T3 (p = 0.005). The noncoding mRNA (DD3/PCA3) was overexpressed 140-fold (p = 0.007) in the cancer samples compared to BPH tissues. Hepsin was overexpressed 21-fold (p = 0.049, whereas the overexpression for PSMA was 66-fold (p = 0.047). When the gene expression data for these 4 biomarkers was combined in a logistic regression model, a predictive index was obtained that distinguished 100% of the PCa samples from all of the BPH samples. Therefore, combining these genes in a real-time PCR assay represents a powerful new approach to diagnosing PCa by molecular profiling. (Supplemental material for this article can be found on the International Journal of Cancer website at http://www.interscience.wiley.com/jpages/0020-7136/suppmat/index.html.)     

Epidermal growth factor and transforming growth factor alpha concentrations in BPH and cancer of the prostate: their relationships with tissue androgen levels.

We measured immunoreactive EGF and TGF alpha in prostate tissue extracts obtained from 19 patients with benign prostatic hyperplasia (BPH) and 19 with cancer of the prostate (CaP). Whilst both BPH and CaP expressed EGF (BPH = 195.61 +/- 19.94 ng g-1 protein; CaP = 235.60 +/- 24.45 ng g-1 protein) and TGF alpha (BPH = 92.57 +/- 7.60 ng g-1 protein; CaP = 100.73 +/- 15.47 ng g-1 protein) in equal concentrations, the levels of EGF in any tissue extract were on average twice those of TGF alpha. Furthermore analysis of the individual growth factor data revealed a direct correlation between EGF and TGF alpha in both BPH (r = 0.72, P < 0.001) and CaP (r = 0.69, P < 0.001). When the tumours were classified according to their Gleason score, a slight but significant increase in growth factor concentrations was noted as the tumour became less differentiated. We also measured the concentrations of testosterone and dihydrotestosterone (DHT) in prostate extracts with a view of elucidating the relationship between androgen and growth factors in this gland. There was a small positive correlation only between testosterone and EGF (r = 0.62, P < 0.05) and testosterone and TGF alpha (r = 0.61, P < 0.05) in CaP. The absence of any similar correlation in BPH where DHT becomes the predominant hormone may suggest an indirect role for testosterone in the regulation of growth factor production.     

Prostate stem cell antigen (PSCA) expression in human prostate cancer tissues: implications for prostate carcinogenesis and progression of prostate cancer

OBJECTIVE: Prostate stem cell antigen (PSCA) is a recently defined homolog of the Thy-1/Ly-6 family of glycosylphosphatidylinositol (GPI)-anchored cell surface antigens. The objective of the present study was to examine the expression status of PSCA protein and mRNA in clinical specimens of human prostate cancer (PCa) and to validate it as a potential molecular target for diagnosis and treatment of PCa. METHODS: Immunohistochemical (IHC) and in situ hybridization (ISH) analyses of PSCA expression were simultaneously performed on paraffin-embedded sections of 20 benign prostatic hyperplasia (BPH), 20 prostatic intraepithelial neoplasm (PIN) and 48 prostate cancer (PCa) tissues, including 9 androgen-independent prostate cancers. The level of PSCA expression was semiquantitatively scored by assessing both the percentage and intensity of PSCA-positive staining cells in the specimens. We then compared the PSCA expression between BPH, PIN and PCa tissues and analyzed the correlations of PSCA expression level with pathological grade, clinical stage and progression to androgen-independence in PCa. RESULTS: In BPH and low grade PIN, PSCA protein and mRNA staining were weak or negative and less intense and uniform than that observed in high grade PIN (HGPIN) and PCa. Moderate to strong PSCA protein and mRNA expression were noted in 8 of 11 (72.7%) HGPIN and in 40 of 48 (83.4%) PCa specimens examined by IHC and ISH analyses, and their statistical significance was compared with BPH (20%) and low-grade PIN (22.2%) specimens (P < 0.05). The expression level of PSCA increased with a higher Gleason grade, advanced stage and progression to androgen-independence (P < 0.05). In addition, IHC and ISH staining revealed a high degree of correlation between PSCA protein and mRNA overexpression. CONCLUSIONS: Our data demonstrate that PSCA as a new cell surface marker is overexpressed in a majority of cases of human PCa. PSCA expression correlates positively with adverse tumor characteristics, such as increasing pathological grade (poor cell differentiation), worsening clinical stage and androgen-independence and speculatively with prostate carcinogenesis. PSCA may possess prognostic utility and may be a promising molecular target for diagnosis and treatment of PCa.     

Increased expression of fibroblast growth factor 13 in prostate cancer is associated with shortened time to biochemical recurrence after radical prostatectomy

Fibroblast growth factor homologous factors (FHFs) belong to the fibroblast growth factor (FGF) superfamily, which plays an important role in prostate cancer (PCa). Mining of public database suggests that FGF13 (FHF2) mRNA expression is altered in over 30% of PCa cases. This study examined the FGF13 expression pattern in human PCa specimens and evaluated its potential as a biomarker for patient outcome after radical prostatectomy (RP). Immunohistochemistry (IHC) showed that FGF13 was detectable in the majority of human PCa samples, and FGF13 IHC scores were higher in high‐grade prostatic intraepithelial neoplasia, in primary PCa and in metastatic PCa than in benign prostatic tissue. There was a significant association between high cytoplasmic FGF13 staining and a risk of biochemical recurrence (BCR) after RP. This was also evident in the intermediate to high‐risk patient groups. In contrast, positive nuclear FGF13 staining along with low cytoplasmic FGF13 group showed a decreased BCR risk. Multivariate regression analysis indicated that high cytoplasmic FGF13 staining was associated with BCR and that this could serve as an independent prognostic marker in PCa. Several PCa cell lines showed increased FGF13 expression at the mRNA and protein levels compared to the immortalized prostate epithelial cell line PNT1a. Analysis of co‐labeled cells suggested a possible interaction of FGF13 with α‐tubulin and the voltage‐gated sodium channel proteins (Na_Vs/VGSCs). Our data indicate that, for PCa patients after RP, FGF13 serves as a potential novel prognostic marker that improves prediction of BCR‐free survival, in particular if combined with other clinical parameters.     

Differential blood-based diagnosis between benign prostatic hyperplasia and prostate cancer: miRNA as source for biomarkers independent of PSA level,Gleason score,or TNM status

Since the benefit of prostate-specific antigen (PSA) screening remains controversial, new non-invasive biomarkers for prostate carcinoma (PCa) are still required. There is evidence that microRNAs (miRNAs) in whole peripheral blood can separate patients with localized prostate cancer from healthy individuals. However, the potential of blood-based miRNAs for the differential diagnosis of PCa and benign prostatic hyperplasia (BPH) has not been tested. We compared the miRNome from blood of PCa and BPH patients and further investigated the influence of the tumor volume, tumor-node-metastasis (TNM) classification, Gleason score, pretreatment risk status, and the pretreatment PSA value on the miRNA pattern. By microarray approach, we identified seven miRNAs that were significantly deregulated in PCa patients compared to BPH patients. Using quantitative real time PCR (qRT-PCR), we confirmed downregulation of hsa-miR-221* (now hsa-miR-221-5p) and hsa-miR-708* (now hsa-miR-708-3p) in PCa compared to BPH. Clinical parameters like PSA level, Gleason score, or TNM status seem to have only limited impact on the overall abundance of miRNAs in patients’ blood, suggesting a no influence of these factors on the expression of deregulated miRNAs.     

Predictive Value of the Platelet-To-Lymphocyte Ratio in Diagnosis of Prostate Cancer

Purpose: To predict prostatic carcinoma using a logistic regression model on prebiopsy peripheral bloodsamples. Materials and Methods: Data of a total of 873 patients who consulted Urology Outpatient Clinics of FatihSultan Mehmet Training and Research Hospital between February 2008 and April 2014 scheduled for prostatebiopsy were screened retrospectively. PSA levels, prostate volumes, prebiopsy whole blood cell counts, neutrophiland platelet counts, neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), biopsy resultsand Gleason scores in patients who had established diagnosis of prostate cancer (PCa) were evaluated. Results:This study was performed on a total of 873 cases, with an age range 48-76 years, divided into three groups asfor biopsy results. with diagnoses of benign prostatic hyperplasia (BPH) (n=304, 34.8 %), PCa (n=265, 30.4 %)and histological prostatitis (n=304; 34.8 %). Intra- and intergroup comparative evaluations were performed.White blood cell and neutrophil counts in the histological prostatitis group were significantly higher than thoseof the BPH and PCa groups (p=0.001; p=0.004; p<0.01). A statistically significant intergroup difference wasfound for PLR (p=0.041; p<0.05) but not lymphocyte count (p>0.05). According to pairwise comparisons, PLRwere significantly higher in the PCa group relative to BPH group (p=0.018, p<0.05, respectively). Though notstatistically significant, higher PLR in cases with PCa in comparison with the prostatitis group was remarkable(p=0.067, and p>0.05, respectively). Conclusions: Meta-analyses showed that in patients with PSA levels over4 ng/ml, positive predictive value of PSA is only 25 percent. Therefore, novel markers which can both detectclinically significant prostate cancer, and also prevent unnecessary biopsies are needed. Relevant to this issuein addition to PSA density, velocity, and PCA3, various markers have been analyzed. In the present study, PLRw ere found to be the additional predictor of prostatic carcinoma.     

Matrix metalloproteinases 1 and 3, tissue inhibitor of metalloproteinase-1 and the complex of metalloproteinase-1/tissue inhibitor in plasma of patients with prostate cancer

We analyzed blood plasma concentrations of matrix metalloproteinase-1 and -3 (MMP-1; MMP-3), the tissue inhibitor of metalloproteinase-1 (TIMP-1) and the complex MMP-1/TIMP-1, and looked for any correlation with prostate cancer stage. These components were measured by ELISA tests specific for these proteins in healthy male controls (n = 35), and in patients with benign prostatic hyperplasia (BPH; n = 29), with prostate cancer (PCa) without metastasis (T2,3pN0M0; n = 29) and with PCa with metastatic disease (T2,3,4pN1,2M1; n = 18). Mean values of MMP-1 and of the complex MMP-1/TIMP-1 were not different among the 4 groups studied. The mean MMP-3 and especially TIMP-1 concentrations were significantly higher in PCa patients with metastases compared with controls, BPH and PCa patients without metastases. Ten of these 18 patients had TIMP-1 concentrations higher than the upper reference limit. TIMP-1 concentrations were correlated with staging but not with grading. Our results point towards plasma TIMP-1 concentration as a potential marker of malignant progression of PCa. Int. J. Cancer 74:220-223, 1997. © 1997 Wiley-Liss, Inc.     

Comparative Expression of RAGE and SOX2 in Benign and Malignant Prostatic Lesions

Background: Prostate cancer (PCa) is a common health problem in elderly. RAGE (Receptor for advanced glycationend products) is overexpressed in multiple human cancers. SOX2 (Sex-determining region Y box 2) also functions as anoncoprotein and promotes cancer progression but the mechanisms involved remain largely unknown. Aim: The currentstudy investigated the expression patterns of RAGE and SOX2 in benign and malignant prostate samples in correlationwith the histopathological findings in order to evaluate their role as prognostic markers or therapeutic targets. Methods:Immunohistochemical staining for RAGE and SOX2 antibodies was applied on 87 prostatic biopsies [16 of prostatitis, 20of benign prostatic hyperplasia (BPH) and 51 of PCa]. Results: Expression of RAGE and SOX2 (percentage of positivecells) was significantly higher in PCa lesions compared with prostatitis (p<0.01) and BPH (p<0.0001) and was alsosignificantly higher in prostatitis compared with BPH lesions (p<0.01). Also, percentage of positive RAGE and SOX2cells showed a significant stepwise increase from Gleason Grade 3 to Grade 5 and were significantly higher in highGleason Scores (≥8) compared to lower Scores (≤7) with statistical significance (p=0.001). Conclusion: RAGE andSOX2 were up-regulated in prostate cancer lesions, mainly in advanced grades, suggesting an active role of both antigensin the development and progression of prostate cancer and expecting the possibility of their use as therapeutic targets.     

Plasma Vascular Endothelial Growth Factors A and C inPatients undergoing Prostatic Biopsy and TURP for SuspectedProstatic Neoplasia

Background: Formation of new blood vessels is necessary for the development and spread of neoplasms morethan 1 mm3 in volume, angiogenesis being responsible for formation of new from pre-existing blood vessels.Vascular endothelial growth factor (VEGF) is pivotal and the best studied angiogenic factor in all humancancers. Therefore we designed this study to investigate the role of VEGF-A and VEGF-C in prostate cancer incomparison with BPH controls in a north Indian population. Methods: In this case-control study a total of 100subjects were included on the basis of confirmed histopathological reports, out of which 50 were prostate cancerpatients and the other 50 were BPH patients with PSA levels >2 ng/ml and abnormal digital rectal examination(DRE) findings during September 2009 to August 2011 from the Department of Urology, KGMU, Lucknow, India.Plasma levels of VEGF were determined using quantitative immunoassay (ELISA- enzyme linked immunosorbentassay). Statistical analysis was carried out using SPSS 15.0 version. Results: The mean age of prostate cancer(67.6±5.72) patients was significantly higher (p=0.005) than BPH (63.6±7.92) patients. Expression of VEGF-Awas not significantly higher in disease stage C1 than D1 or D2 and A or B (p=0.13) while the level of VEGF-Awas significantly higher (p=0.04) in prostate cancer as compared to BPH subjects (PCa=13.0 pg/ml, BPH=6.8pg/ml). Levels of VEGF-C were similar in both groups (PCa=832.6 pg/ml, BPH=823.7 pg/ml). In ROC curve,the area under curve (AUC) was 0.70 (95%CI: 0.60-0.80) and the cut-off value for which a higher proportion ofpatients was correctly classified (20%) was 26.0 pg/mL. Conclusion: Although VEGF-A is increased in cancerprostate patients a statistically significant correlation could not be established in this study. VEGF-C was notfound to be a useful biomarker.     

Frequency and Type Distribution of Human Papilloma Virus in Patients with Prostate Cancer,Kerman, Southeast of Iran

Prostatic cancer is the second cause of cancer-related death among men worldwide. The human papilloma viruses (HPVs) are a family of sexually transmitted viruses which have may have roles in the ethiology of in ammation in prostate leading to benign prostatic hyperplasia (BPH) and prostate cancer (PCa). In this study, we evaluated the frequency of different HPV types in prostatic cancer and benign prostatic hyperplasia (BPH) in Kerman province, southeast of Iran, using real-time PCR techniques. The aim of the present research was to clarify any association with prostatic carcinogenesis. Real Time PCR showed that HPV DNA was found in 20% of 200 PCa samples, 80 percent of these with high-risk HPV types, 40% with type-16,18, 30 % type-31,33 and 10% type 54. High risk HPV DNA was detected in only 2% of BPH samples. Values for low risk types were much higher. Our study provided a support for the role of high risk HPV infection in prostatic disease in Iranian patients, and association between presence of HPV DNA and prostate carcinoma. In particular, HPV 16 and18 might have an important role in prostate cancer.     

Expression and significance of microsomal prostaglandin synthase-1 (mPGES-1) and Beclin-1 in the development of prostate cancer

The aim of this study was to investigate the expression and significance of microsomal prostaglandin synthase-1 (mPGES-1) and Beclin-1 in the development of prostate cancer (PCa). Immunohistochemistry was performed on paraffin-embedded sections with rabbit polyclonal against mPGES-1 and Beclin-1 in 40 PCa, 40 benign prostatic hyperplasia (BPH) and 10 normal prostate specimens for this purpose. Quantitative real-time polymerase chain reaction (qRT-PCR) was applied for mRNA expression of mPGES-1 and Beclin-1, while MTT assays were used to ascertain the best working concentration of the mPGES-1 inhibitor (CAY10526). The effect of CAY10526 treatment on expression of Beclin-1 in DU-145 cells was studied using Western blot analysis. Localization of Beclin-1 and mPGES-1 was in endochylema. Significant differences in expression was noted among PCa, BPH and normal issues (P<0.05). Beclin-1 expression inversely correlated with mPGES-1 expression in PCa tissue (P<0.05). CAY10526 could significantly block mPGES-1 expression and the proliferation of DU-145 cells (P<0.05), while increasing Beclin-1 levels (P<0.05). Overexpression of mPGES-1 could decrease the autophagic PCa cell death. Inhibiting the expression of mPGES-1 may lead to DU-145 cell death and up-regulation of Beclin-1. The results suggest that inhibition of mPGES-1 may have therapeutic potential for PCa in the future.