PCA3 mRNA Research Articles

Diagnostic potential value of circulating PCA3 mRNA in plasma and urine of prostate cancer patients

Current reports show that most of the routinely available prostate specific antigen (PSA) derived biomarkers of prostate cancer (PCa) have failed to demonstrate sufficient sensitivity and specificity in clinical use for diagnosis and prognosis of PCa. Consequently, a number of molecular markers including the prostate cancer gene 3 (PCA3) are being considered for clinical use. This study was therefore designed to determine the levels of PCA3 mRNA expression in urine and plasma of prostate cancer patients and to assess its diagnostic characteristics in Nigerian population with prostate cancer. A total of 90 participants comprising 30 participants each with PCa, BPH and 30 apparently healthy adults as controls were enrolled into this case-control study. Blood and urine samples were collected from study participants and dispensed into PaxGene RNA protecting fluid, lithium heparin bottles and nuclease free bottle as appropriate. Anthropometric indices were obtained using standard methods, while the plasma levels of PSA, testosterone and estrogen were determined using enzyme linked immunosorbent assay (ELISA). The PCA3 mRNA expression in urine and plasma was determined using Quantitative Polymerase Chain Reaction. Comparison of groups was done using analysis of variance (ANOVA) followed by Tukey Post-Hoc test. The sensitivity and specificity of PCA3 mRNA in the diagnosis of PCa was evaluated through estimation of the area under the receiver operating characteristic (ROC) curve. 'The mean waist circumference (WC) and (HC) was significantly higher in the BPH group (43.94 ± 2.32) compared to both the control (36.04 ± 0.80) and the PCa group (38.50 ± 0.94) (p < 0.05). The prostate cancer group had a significantly higher levels (p < 0.05) of tPSA (47.58 ± 12.23) and an increased expression of PCA3 mRNA in urine (10.46 ± 1.68) and plasma (14.89 ± 1.37) when compared with control values [tPSA (0.67 ± 0.27), urine (2.22 ± 0.22) and plasma (1.45 ± 0.13) expression of PCA3 mRNA]. A negative correlation was found to exist between Plasma PCa3 mRNA and plasma levels of estradiol (r=-0.425, p = 0.024) and testosterone (r=-0.423, p = 0.025) in PCa group. A similar trend was observed between urine PCA3 mRNA, estrogen (r=-0.459, p = 0.014) and testosterone (r=-0.407, p = 0.031). The ROC curve shows the tPSA (p = 0.03) is the most sensitive marker of detection followed by plasma PCA3 mRNA and subsequently the urine PCA3 mRNA. The curve further shows the urine PCA3 mRNA (p = 0.006) is the most specific for detection of prostate cancer and the less and least sensitive are the plasma PCA3 mRNA and tPSA. It could be concluded from this study that there is reduced levels of estradiol and testosterone and elevated levels of tPSA and PCA3 mRNA expression in urine and blood in patients with benign prostatic hyperplasia and prostate cancer. Hence, determination of plasma or urine expression of PCA3 mRNA is valueable in the diagnosis and treatment monitoring of prostate cancer.

Prostatic aspirated cellular RNA analysis enables fast diagnosis and staging of prostate cancer

ObjectiveThe aim of this study is to investigate the potential application of prostatic aspirated cellular RNA analysis technique for fast diagnosing and staging prostate cancer. MethodsProstatic aspirated cells were obtained immediately after transrectal ultrasound (TRUS)-guided needle biopsy. Cellular RNA such as glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA, prostate specific antigen (PSA) mRNA and prostate-specific RNA (PCA3) mRNA were analyzed by using Reverse Transcription-Polymerase Chain Reaction (RT-PCR). PCA3 score was calculated as the ratio of PCA3 mRNA to PSA mRNA expression. Diagnostic performance of the fast-aspirated cellular RNA analysis technique for determining prostate cancer and metastatic status were evaluated by developing receiver operating characteristic curves (ROC), and the correlation between aspirated cellular RNA mRNA expressions and risk grouping was calculated, to investigate the underlying potential for PCa staging. ResultsPCA3 score was significantly higher in prostatic aspirated cells obtained from cancerous tissues than noncancerous tissues. The median aspirated cellular PCA3 score was higher in patients with PCa compared to BPH, and presenting an area under the ROC curve (AUC) of 0.87 (95%CI: 0.79–0.94) for PCa diagnosis. Multivariate regression analysis revealed that baseline median aspirated cellular PCA3 score (OR=9.316, 95%CI: 1.045–83.033, P<0.05) was an independent predictive factor for metastatic status in PCa patients. ConclusionThe ease of use and minimal complexity of fast prostatic aspirated cellular RNA analysis may be a valuable diagnostic technique, providing urgent diagnostic information for accurate PCa diagnosing and staging.

Identification of Clinically Significant Prostate Cancer by Combined PCA3 and AMACR mRNA Detection in Urine Samples.

PurposePreclinical evaluation of PCA3 and AMACR transcript simultaneous detection in urine to diagnose clinical significant prostate cancer (prostate cancer with Gleason score ≥7) in a Russian cohort.Patients and MethodsWe analyzed urine samples of patients with a total serum PSA ≥2 ng/mL: 31 men with prostate cancer scheduled for radical prostatectomy, 128 men scheduled for first diagnostic biopsy (prebiopsy cohort). PCA3, AMACR, PSA and GPI transcripts were detected by multiplex reverse transcription quantitative polymerase chain reaction, and the results were used for scores for calculation and statistical analysis.ResultsThere was no significant difference between clinically significant and nonsignificant prostate cancer PCA3 scores. However, there was a significant difference in the AMACR score (patients scheduled for radical prostatectomy p=0.0088, prebiopsy cohort p=0.029). We estimated AUCs, optimal cutoffs, sensitivities and specificities for PCa and csPCa detection in the prebiopsy cohort by tPSA, PCA3 score, PCPT Risk Calculator and classification models based on tPSA, PCA3 score and AMACR score. In the clinically significant prostate cancer ROC analysis, the PCA3 score AUC was 0.632 (95%CI: 0.511–0.752), the AMACR score AUC was 0.711 (95%CI: 0.617–0.806) and AUC of classification model based on the PCA3 score, the AMACR score and total PSA was 0.72 (95%CI: 0.58–0.83). In addition, the correlation of the AMACR score with the ratio of total RNA and RNA of prostate cells in urine was shown (tau=0.347, p=6.542e–09). Significant amounts of nonprostate RNA in urine may be a limitation for the AMACR score use.ConclusionThe AMACR score is a good predictor of clinically significant prostate cancer. Significant amounts of nonprostate RNA in urine may be a limitation for the AMACR score use. Evaluation of the AMACR score and classification models based on it for clinically significant prostate cancer detection with larger samples and a follow-up analysis is promising.

The Percentage of Free PSA and Urinary Markers Distinguish Prostate Cancer from Benign Hyperplasia and Contribute to a More Accurate Indication for Prostate Biopsy.

The main advantage of urinary biomarkers is their noninvasive character and the ability to detect multifocal prostate cancer (CaP). We have previously implemented a quadruplex assay of urinary markers into clinical practice (PCA3, AMACR, TRPM8 and MSMB with KLK3 normalization). In this study, we aimed to validate it in a larger cohort with serum PSA 2.5–10 ng/mL and test other selected transcripts and clinical parameters, including the percentage of free prostate-specific antigen (PSA) (% free PSA) and inflammation. In the main cohort of 299 men, we tested the quadruplex transcripts. In a subset of 146 men, we analyzed additional transcripts (CD45, EPCAM, EZH2, Ki67, PA2G4, PSGR, RHOA and TBP). After a prostate massage, the urine was collected, RNA isolated from a cell sediment and qRT-PCR performed. Ct values of KLK3 (i.e., PSA) were strongly correlated with Ct values of other genes which play a role in CaP (i.e., PCA3, AMACR, TRPM8, MSMB and PSGR). AMACR, PCA3, TRPM8 and EZH2 mRNA expression, as well as % free PSA, were significantly different for BPH and CaP. The best combined model (% free PSA plus PCA3 and AMACR) achieved an AUC of 0.728 in the main cohort. In the subset of patients, the best AUC 0.753 was achieved for the combination of PCA3, % free PSA, EPCAM and PSGR. PCA3 mRNA was increased in patients with inflammation, however, this did not affect the stratification of patients indicated for prostate biopsy. In conclusion, the percentage of free PSA and urinary markers contribute to a more accurate indication for prostate biopsy.

&lt;p&gt;Plasma Messenger RNAs Identified Through Bioinformatics Analysis are Novel, Non-Invasive Prostate Cancer Biomarkers&lt;/p&gt;

AimTo identify new biomarkers of prostate cancer (PCa) for the diagnosis and prediction of clinical outcomes.Materials and MethodsExisting microarray data of PCa tissues in the Oncomine database were analyzed and candidate differentially expressed genes (DEGs) that may be novel and noninvasive biomarkers were obtained. On this basis, plasma mRNA was extracted from PCa patients and healthy donors. Furthermore, plasma mRNA expression of DEGs was evaluated by qRT-PCR. Finally, the diagnostic power of the biomarkers was evaluated in comparison to the clinical characteristics of the patients.ResultsIn this study, the top five significantly overexpressed mRNA (AMACR, PPP1R14b, PCA3, DLX1, and RPL22L1) and the top five significantly underexpressed mRNA (DUOX1, EFS, GSTP1, S100A16, and NCRNA00087) were selected for further validation in PCa patients and healthy donors by qRT-PCR. The results showed that AMACR, DLX1, PCA3, DUOX1, and GSTP1 mRNA were stably amplified in plasma. Additionally, DLX1, PCA3, DUOX1, and GSTP1 mRNA expression was significantly different between PCa circulating free mRNA samples and healthy donors. These mRNAs may be useful biomarkers for PCa diagnosis.ConclusionAnalysis of the expression of genes in the Oncomine database showed that DLX1, PCA3, and DUOX1 expressions have a cancer specific pattern in PCa. Collectively, DLX1, PCA3, and DUOX1 may be useful candidate biomarkers for PCa diagnosis.

Перспективы совершенствования диагностики рака предстательной железы на основе анализа экспрессии гена РСА3

Prostate cancer is the second leading cause of cancer death. The widespread introduction into the clinical practice of test for prostate specific antigen (PSA) led to an increase in the number of prostate biopsies performed. At the same time, a decrease in the threshold of age-specific PSA standards has resulted in an increase in the number of unnecessary biopsies. In this regard, a need has arisen for new prostate cancer biomarkers. PCA3 is a non-coding mRNA that is exclusively expressed by prostate cells. Currently, three generations of test diagnostic systems based on the quantitative analysis of the PCA3 mRNA in the urine or its cell sediment has already developed, and they differ in the type of material studied and the method for estimating the amount of PCA3 mRNA. Clinical studies of the developed test systems have shown that a high level of PCA3 expression in the patients urine correlates with the probability of detecting prostate cancer. PCA3 test has higher positive and negative predictive values than previously used PSA test. These data are repeatedly confirmed by studies conducted in different clinics. Thus, the introduction of the method of quantitative determination of PCA3 in clinical practice can significantly improve the efficiency of diagnosis of prostate cancer and reduce the number of unnecessary biopsies.

Ultrasensitive and Reversible Nanoplatform of Urinary Exosomes for Prostate Cancer Diagnosis.

Prostate cancer cell-derived exosomes in urine have been extensively studied recently and regarded as novel biomarkers for cancer diagnosis and prognosis, which presents wide prospects in clinical applications. Sensitive detection and specific capture methods are essential for exosomes analysis. Herein, a dual functional platform composed of superparamagnetic conjunctions and molecular beacons (SMC-MB) is reported. The SMC-MB platform is designed based on aptamer immunoaffinity with ultrasensitive detection efficiency and reversible isolation capacity, which, respectively, profit from nonenzymatic amplification methods and magnetic separation along with restriction cleavage. It is noteworthy that exosomes quantification was exactly amplified and transformed into single strand DNA detection. Correlated measurements evidence that the limit of detection of SMC-MB is as low as ∼100 particles/μL in urine, and a linear relationship meets between the logarithmic concentration of exosomes and fluorescence intensity of the molecular beacon. Furthermore, employing prostate specific membrane antigen (PSMA) aptamer, the platform adapted to detect and capture PMSA-positive exosomes from urine samples provides excellent diagnostic efficiency for prostate cancer (PCa). The expression of typical biomarkers of PCa, i.e., PSA and PCA3 mRNA, is significantly higher in PSMA-positive exosomes. Altogether, the platform and strategy described in this paper are promising in urinary exosomes analysis and prostate cancer detection.

Urine PCA3 mRNA level in diagnostic of prostate cancer.

The objective of this study is to evaluate the diagnostic effects of urine PCA3 mRNA level in patients with prostate cancer (PC). Twenty four patients with pathologically confirmed PC, 40 patients with benign prostatic hyperplasia (BPH), and 13 patients with urolithiasis were recruited in this study. The urine level of PCA3 mRNA was determined with real-time polymerase chain reaction and compared among the three groups. With the reference parameter of urine PCA3 mRNA, the diagnostic sensitivity, specificity, and area under the receiver operating characteristic curve (AUC) were calculated. The mRNA level in urine of PC, BPH and urolithiasis were 60.2 ± 32.2, 17.1 ± 12.9, and 6.2 ± 3.1, respectively, indicating that mRNA level in PC was significantly higher than that of in other two groups (P < 0.05). The AUC was 0.90 while the sensitivity and specificity were 87.5% and 79.2% with the cutoff value of 33.86. The urine PCA3 mRNA could be a biomarker for diagnosing patients with PC.

Evaluation of Combined Quantification of PCA3 and AMACR Gene Expression for Molecular Diagnosis of Prostate Cancer in Moroccan Patients by RT-qPCR

Prostate cancer (PCa) remains one of the most widespread and perplexing of all human malignancies. Assessment of gene expression is thought to have an important impact on cancer diagnosis, prognosis and therapeutic decisions. In this context, we explored combined expression of PCa related target genes AMACR and PCA3 in 126 formalin fixed paraffin embedded prostate tissues (FFPE) from Moroccan patients, using quantitative real time reverse transcription-PCR (RT-qPCR). This quantification required data normalization accomplished using stably expressed reference genes (RGs). A panel of twelve RG was assessed, data being analyzed using GenEx V6 based on geNorm, NormFinder and statistical methods. Accordingly, the hnRNP A1 gene was identified and selected as the most stably expressed RG for reliable and accurate gene expression quantification in prostate tissues. The ratios of both PCA3 and AMACR gene expression relative to that of the hnRNP A1 gene were calculated and the performance of each target gene for PCa diagnosis was evaluated using receiver-operating characteristics. PCA3 and AMACR mRNA quantification based on RT-qPCR may prove useful in PCa diagnosis. Of particular interesting, combining PCA3 and AMACR quantification improved PCa prediction by increasing sensitivity with retention of good specificity.

Altered PCA3 and TMPRSS2-ERG expression in histologically benign regions of cancerous prostates: a systematic, quantitative mRNA analysis in five prostates.

BackgroundPCA3 and TMPRSS2-ERG are commonly overexpressed biomarkers in prostate cancer, but reports have emerged demonstrating altered expression also in areas outside the tumour foci in cancerous prostates. Our aim was to measure PCA3 and TMPRSS2-ERG expression systematically in all regions of prostate cross-sections, matching the data to corresponding tissue morphology.MethodsTMPRSS2-ERG and PCA3 mRNA levels were measured with quantitative reverse-transcription PCR assays in 270 samples from cross-sections of five radical prostatectomy specimens. ERG expression was examined by immunohistochemistry.ResultsTMPRSS2-ERG mRNAs were detected in three patients and in 15 tissue samples in total. These included two carcinoma samples and 13 histologically benign samples, eight of which were located next to malignant tumours or PIN (prostatic intraepithelial neoplasia) lesions and five of which did not reside in the vicinity of any evident carcinoma foci. ERG protein expression was limited to areas of TMPRSS2-ERG mRNA expression, but did not identify all of them. PCA3 expression was detected in all five cross-sections, with statistically significant, three-fold higher expression in carcinoma regions.ConclusionsTMPRSS2-ERG expression was detected in carcinoma foci, regions next to them, and in samples not adjacent to carcinoma foci. Claimed as a cancer-associated phenomenon, this fusion gene measurement could, if validated with a larger cohort, be utilized as an addition to histological analysis to predict current or future cancer risk in men with negative biopsies. Molecular changes outside the carcinoma foci are also indicated for PCA3, as its expression was only moderately increased in the carcinoma regions.

Expression of PRUNE2 mRNA and its positive correlation with non-coding RNA PCA3 in leiomyosarcoma

Our previous study shows that PURNE2 mRNA plays an important role in the differential diagnosis of leiomyosarcoma and gastrointestinal stromal tumor (GIST). Non-coding RNA PCA3 locates in the intron of PRUNE2 and may play a role in PRUNE2 expression. The aim of this study was to explore the expression of PCA3 mRNA and PRUNE2 in leiomyosarcoma and their correlation. The expression of PRUNE2 mRNA was analyzed by agilent gene expression microarray CHIP in 31 leiomyosarcomas and 37 GISTs, and the correlation of the PRUNE2 expression and prognosis of leiomyosarcoma was predicted. Real-Time PCR assay was used to detect the mRNA levels of PCA3 and PRUNE2 in 13 leiomyosarcomas and to investigate their correlation. Seven prostate cancer tissues were used as control of PCA3. The level of PRUNE2 mRNA expression was significantly higher in the 31 leiomyosarcomas than that in the 37 GISTs, and the level of PRUNE2 mRNA expression was correlated with survival of the leiomyosarcoma patients. Compared with prostate cancer, the non-coding RNA PCA3 expression level was significantly lower in leiomyosarcoma, and it had no correlation with the prognosis of leiomyosarcoma. Most importantly, the PRUNE2 and PCA3 mRNA expressions were both upregulated in leiomyosarcoma and showed a significant positive correlation. Our findings demonstrate for the first time that PRUNE2 expression is correlated with the survival of leiomyosarcoma patients. Furthermore, non-coding RNA PCA3, which locates in the intron of PRUNE2, has a significant positive correlation with PRUNE2 and may play an important role in the pathogenesis of leiomyosarcoma.

KLK3, PCA3, and TMPRSS2-ERG expression in the peripheral blood mononuclear cell fraction from castration-resistant prostate cancer patients and response to docetaxel treatment.

To monitor systemic disease activity, the potential of circulating tumor cells (CTCs) bears great promise. As surrogate for CTCs we measured KLK3, PCA3, and TMPRSS2-ERG messenger RNA (mRNA) in the peripheral blood mononuclear cell (PBMC) fraction from a castration-resistant prostate cancer (CRPC) patient cohort and three control groups. Moreover, biomarker response to docetaxel treatment was evaluated in the patient group. Blood samples from 20 CRPC patients were analyzed at four different time points (prior to docetaxel treatment, at 9 weeks, 27 weeks, and 2 months after treatment). Blood was drawn once from three control groups (10 age-matched men, 10 men under 35 years of age, 12 women). All samples were analyzed for KLK3, PCA3, and TMPRSS2-ERG mRNA by using a quantitative nucleic acid amplification assay with gene-specific primers in the complementary DNA synthesis. At baseline, mRNA for KLK3 was detected in 17 (89%, 95% CI 76-100%), PCA3 in 10 (53%, 95% CI 30-75%), and TMPRSS2-ERG in seven of 19 evaluable patients (37%, 95% CI 15-59%). In contrast, the blood samples from all 32 healthy volunteers were reproducible negative for all markers. In response to docetaxel treatment, KLK3 levels decreased in 80% (95% CI 60-100%), PCA3 in 89% (95% CI 68-100%), and TMPRSS2-ERG in 86% (95% CI 60-100%) of patients. The feasibility of a highly sensitive modified nucleic acid amplification assay to assess KLK3, PCA3, and TMPRSS2-ERG mRNA in the PBMC fraction from CRPC patients was demonstrated. Moreover, response of these markers to systemic treatment was shown.

MP78-08 RACIAL/ETHNIC PATTERNS IN THE PERFORMANCE OF MULTIPLEX URINE ERG AND PCA3 FOR PROSTATE CANCER DETECTION

You have accessJournal of UrologyProstate Cancer: Epidemiology & Natural History III1 Apr 2014MP78-08 RACIAL/ETHNIC PATTERNS IN THE PERFORMANCE OF MULTIPLEX URINE ERG AND PCA3 FOR PROSTATE CANCER DETECTION Yongmei Chen, Jennifer Cullen, Aminia Ali, David G. McLeod, and Shiv Srivastava Yongmei ChenYongmei Chen More articles by this author , Jennifer CullenJennifer Cullen More articles by this author , Aminia AliAminia Ali More articles by this author , David G. McLeodDavid G. McLeod More articles by this author , and Shiv SrivastavaShiv Srivastava More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2014.02.2491AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Introduction: Considering the limitations of PSA in prostate cancer (CaP) detection, a number of noninvasive and high specific tests have been developed for improving the prediction accuracy (PA) of CaP diagnosis. However, the racial/ethnic interactions with these biomarkers remain poorly understood. This study reports on the racial/ethnic patterns in the performance of urine ERG and PCA3 for CaP detection. METHODS Material and Methods: Data were collected from 306 men who underwent TRUS 12 core prostate biopsy. PCA3-, ERG-, and PSA-mRNA concentrations were quantified using the DTS® 400 system assay derived from first-catch urine samples following a DRE. The association of ERG score with clinical parameters were evaluated in whole biopsy cohort, as well as stratified by race/ethnicity; Secondly, PA estimates of biopsy result were quantified using area under the curve (AUC) of the receiver operator characteristic (ROC) analysis in multivariate logistic regression models, and assessed using cross-validation, in further, repeat these analyses in racial groups separately; Finally, quartile cutoffs were adopted to assess cancer risk in white and black men with extremes of ERG and PCA3 scores, respectively. RESULTS Of the 306 men in the study, 174 (57%) were diagnosed with CaP; 116 (38%) self-reported as black race/ethnicity. Median ERG score was significantly higher in white men (59 vs. 30, p = 0.03). Men diagnosed with CaP had much higher ERG score (67 vs. 24, p < .001) than men with a negative biopsy. This difference was widened among white biopsy group (85 vs. 20, p < .001). This difference in ERG score was not observed in black biopsy group (29 vs.32, p = 0.84). The combination of ERG and PCA3 score had a significant increased 15% of AUC as compared to serum PSA alone (0.68 vs. 0.53, p = 0.0062) for predicting CaP in white men. In contrast, the biomarker panel performed more poorly than serum PSA alone in black patients (AUC = 0.49 vs. 0.61, p = 0.0770). CONCLUSIONS Post-DRE urine ERG and PCA3 mRNA expression levels were identified as novel and informative markers for increasing the PA for CaP diagnosis among white men. This panel, in combination with race/ethnicity and PSA, may provide an opportunity to help patients and physicians make informed decisions about early detection, biopsy, and management of prostate cancer. © 2014FiguresReferencesRelatedDetails Volume 191Issue 4SApril 2014Page: e923 Advertisement Copyright & Permissions© 2014MetricsAuthor Information Yongmei Chen More articles by this author Jennifer Cullen More articles by this author Aminia Ali More articles by this author David G. McLeod More articles by this author Shiv Srivastava More articles by this author Expand All Advertisement Advertisement PDF DownloadLoading ...

Expressed Prostatic Secretion Biomarkers Improve Stratification of NCCN Active Surveillance Candidates: Performance of Secretion Capacity and TMPRSS2:ERG Models

Active surveillance is a viable patient option for prostate cancer provided that a clinical determination of low risk and presumably organ confined disease can be made. To standardize risk stratification schemes the NCCN (National Comprehensive Cancer Network®) provides guidelines for the active surveillance option. We determined the effectiveness of expressed prostatic secretion biomarkers for detecting occult risk factors in NCCN active surveillance candidates. Expressed prostatic secretion specimens were obtained before robot-assisted radical prostatectomy. Secretion capacity biomarkers, including total RNA and expressed prostatic secretion specimen volume, were measured by standard techniques. RNA expression biomarkers, including TXNRD1 mRNA, prostate specific antigen mRNA, TMPRSS2:ERG fusion mRNA and PCA3 mRNA, were measured by quantitative reverse-transcription polymerase chain reaction. Of the 528 patients from whom expressed prostatic secretions were collected 216 were eligible for active surveillance under NCCN guidelines. Variable selection on logistic regression identified 2 models, including one featuring types III and VI TMPRSS2:ERG variants, and one featuring 2 secretion capacity biomarkers. Of the 2 high performing models the secretion capacity model was most effective for detecting cases in this group that were up-staged or up-staged plus upgraded. It decreased the risk of up-staging in patients with a negative test almost eightfold and decreased the risk of up-staging plus upgrading about fivefold while doubling the prevalence of up-staging in the positive test group. Noninvasive expressed prostatic secretion testing may improve patient acceptance of active surveillance by dramatically reducing the presence of occult risk factors among those eligible for active surveillance under NCCN guidelines.

Разработка диагностической тест-системы для ранней неинвазивной диагностики рака простаты, основанной на количественной детекции мРНК гена PCA3 в осадке мочи методом ОТ-ПЦР в режиме реального времени

The wide introduction of prostatic specific antigen (PSA) determination into clinical practice has resulted in a larger number of prostate biopsies, while the lower age threshold for PSA has led to a larger number of unnecessary prostate biopsies. Hence, there is a need for new biomarkers that can detect prostate cancer. PCA3 is a noncoding messenger ribonucleic acid (mRNA) that is expressed exclusively in prostate cells. The aim of the study has been to develop a diagnostic test system for early non-invasive detection of prostate cancer based on PCA3 mRNA levels in urine sediment using quantitative reverse transcription polymerase chain reaction (qRT-PCR). As part of the study, a laboratory diagnostic test system prototype has been designed, an application methodology has been developed and specificity and sensitivity data of the method has been assessed. The diagnostic system has demonstrated its ability to detect significantly elevated levels of PCA 3/KLK 3 in samples from prostate cancer (PCa) patients compared with those from healthy men. The findings have shown relatively high diagnostic sensitivity, specificity and negative-predictive values for an early non-invasive screening of prostate cancer

Gene expression of PCA3 in peripheral blood and urine and the significance of urine PCA3 score indiagnosis of prostate cancer

Objective To detect the gene expression of PCA3 and PSA in peripheral blood and urine simultaneously to investigate whether PCA3 combining PSA gene could become new markers for diagnosis of Pca. Methods From June 2009 to December 2009,the initial urine after prostatic massage and the peripheral blood specimens were collected from 37 patients with PCa and 68 patients with BPH that were pathologically confirmed,g patients with urinary stone were used as normal control,the expression of PCA3 and PSA mRNA of mononuclear cells in urine sediments and peripheral blood were detected by fluorescence real-time quantitative PCR,with β-actin mRNA as internal control. Results The sensitivity and specificity of the expression of PCA3 mRNA in peripheral blood for diagnosis of prostate cancer were 48.6％ and 100％ respectively.ROC curve analysis was performed for the PCA3 score and the area under the ROC curve was 0.908.Using 64.6 as the cutoff,the sensitivity was 81.1％ and the specificity was 86.8％.In group with serum tPSA value ＜4 pg/L,the positive rate and negative rate of urinary PCA3 score for diagnosing prostate cancer were 80％ (4/5) and 89.4％ (20/22) respectively.In group with serum tPSA value 4 - 10 μg/L,the positive rate and negative rate of urinary PCA3 score were 66.7％ ( 2/3 ) and 84.2％(16/19) respectively.In group with serum tPSA value ＞ 10 μg/L,the positive rate and negative rate of urinary PCA3 score were 82.8％ (24/27) and 81.5％ (22/27) respectively.The sensitivity of simultaneous detection of PCA3 mRNA in peripheral blood and urinary PCA3 score was 86.5％. Conclusions The expression of PCA3 mRNA in peripheral blood was a specific marker for the diagnosis of PCa.The simultaneous detection of PCA3 mRNA in peripheral blood and urinary PCA3 score could increase the sensitivity for the diagnosis of PCa. Key words: PCA3; PSA; Urinary PCA3 score; PCa; Real-time quantitative PCR

2112 CORRELATION OF URINE PCA3 SCORE AND TMPRS2:ERG SCORE WITH CLINICALLY SIGNIFICANT TUMOR VOLUME IN RADICAL PROSTATECTOMY SPECIMENS: INTERIM RESULTS FROM A MULTI-CENTER PROSPECTIVE STUDY

You have accessJournal of UrologyProstate Cancer: Markers I1 Apr 20122112 CORRELATION OF URINE PCA3 SCORE AND TMPRS2:ERG SCORE WITH CLINICALLY SIGNIFICANT TUMOR VOLUME IN RADICAL PROSTATECTOMY SPECIMENS: INTERIM RESULTS FROM A MULTI-CENTER PROSPECTIVE STUDY John Davis, Patricia Troncoso, Sarah Meyer, John Day, Jack Groskopf, Sheila Aubin, Yvonne Rodriguez, Dan W. Lin, Funda Vakar-Lopez, Cristina Magi-Galluzzi, and Ryan Berglund John DavisJohn Davis Houston, TX More articles by this author , Patricia TroncosoPatricia Troncoso Houston, TX More articles by this author , Sarah MeyerSarah Meyer San Diego, CA More articles by this author , John DayJohn Day San Diego, CA More articles by this author , Jack GroskopfJack Groskopf San Diego, CA More articles by this author , Sheila AubinSheila Aubin San Diego, CA More articles by this author , Yvonne RodriguezYvonne Rodriguez San Diego, CA More articles by this author , Dan W. LinDan W. Lin Seattle, WA More articles by this author , Funda Vakar-LopezFunda Vakar-Lopez Seattle, WA More articles by this author , Cristina Magi-GalluzziCristina Magi-Galluzzi Cleveland, OH More articles by this author , and Ryan BerglundRyan Berglund Cleveland, OH More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2012.02.2280AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Previous studies in men scheduled for prostate biopsy or radical prostatectomy (RP) have shown that urine PCA3 levels correlated with positive versus negative biopsies and total tumor volume in RP specimens (Nakanishi et al J Urol 2008). Given the need for novel markers to select patients for active surveillance, we report interim findings from a large, prospective, multi-institutional validation study. METHODS The primary goal was to correlate PCA3 score with well-characterized tumor volume from men undergoing RP. A secondary goal included assessing the additive prediction from a novel urine assay of TMPRSS2:ERG (T2:ERG). Post-DRE urine samples were collected prior to RP. PCA3 mRNA copies were quantified using a transcription-mediated amplification assay and normalized to PSA mRNA copies to calculate a PCA3 score. T2:ERG mRNA copies were quantified and normalized to PSA mRNA copies to calculate a T2:ERG score. PCA3 score, T2:ERG score and serum PSA were assessed individually and combined for their ability to predict prostatectomy indicators of cancer aggressiveness: significant tumor volume (insignificant tumor volume defined as <0.5cc, Gl ≤ 6, and pT2x), pathologic Gleason Score and non-organ confined disease. RESULTS As of 11/1/2011, 250 of 300 planned participants have been enrolled, and 163 subjects have complete data for interim analysis. Insignificant tumor volume was found in 22 (14%) of 163. The AUC analysis showed statistically significant (P<0.05) power to predict insignificant tumor volume for serum PSA (sPSA), PCA3, and T2:ERG0.72, 0.71, and 0.62 respectively. Other AUC analyses demonstrated statistically significant prediction of Gleason score > 6 by all 3 markers, tumor volume > 0.5 for PCA3 and sPSA, and organ confined disease by only sPSA. At a high cutoff of 70, PCA3 predicted 26% of significant cancer at a specificity of 95% while T2:ERG at a cutoff of 160 predicted 22% of significant cancer at a specificity of 100%. Combining PCA3 and T2:ERG at these cutoffs provided 40% (95% CI, 31 - 49%) prediction of significant cancer at 95% specificity (95% CI, 75–100%). In a logarithmic analysis of sPSA, PCA3, and T2:ERG, the prediction of significant cancer was 60% sensitivity at 90% specificity. CONCLUSIONS In an interim analysis of a validation study, PCA3 and T2:ERG show additive prediction of clinically significant tumor volume, that may be useful in the selection and monitoring of active surveillance candidates. © 2012 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 187Issue 4SApril 2012Page: e852-e853 Advertisement Copyright & Permissions© 2012 by American Urological Association Education and Research, Inc.MetricsAuthor Information John Davis Houston, TX More articles by this author Patricia Troncoso Houston, TX More articles by this author Sarah Meyer San Diego, CA More articles by this author John Day San Diego, CA More articles by this author Jack Groskopf San Diego, CA More articles by this author Sheila Aubin San Diego, CA More articles by this author Yvonne Rodriguez San Diego, CA More articles by this author Dan W. Lin Seattle, WA More articles by this author Funda Vakar-Lopez Seattle, WA More articles by this author Cristina Magi-Galluzzi Cleveland, OH More articles by this author Ryan Berglund Cleveland, OH More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Controversies in using urine samples for prostate cancer detection: PSA and PCA3 expression analysis

Prostate cancer (PCa) is one of the most commonly diagnosed malignancies in the world. Although PSA utilization as a serum marker has improved prostate cancer detection it still presents some limitations, mainly regarding its specificity. The expression of this marker, along with the detection of PCA3 mRNA in urine samples, has been suggested as a new approach for PCa detection. The goal of this work was to evaluate the efficacy of the urinary detection of PCA3 mRNA and PSA mRNA without performing the somewhat embarrassing prostate massage. It was also intended to optimize and implement a methodological protocol for this kind of sampling. Urine samples from 57 patients with suspected prostate disease were collected, without undergoing prostate massage. Increased serum PSA levels were confirmed by medical records review. RNA was extracted by different methods and a preamplification step was included in order to improve gene detection by Real-Time PCR. An increase in RNA concentration with the use of TriPure Isolation Reagent. Despite this optimization, only 15.8% of the cases showed expression of PSA mRNA and only 3.8% of prostate cancer patients presented detectable levels of PCA3 mRNA. The use of a preamplification step revealed no improvement in the results obtained. This work confirms that prostate massage is important before urine collection for gene expression analysis. Since PSA and PCA3 are prostate specific, it is necessary to promote the passage of cells from prostate to urinary tract, in order to detect these genetic markers in urine samples.

Abstract 290: Use of serum and tissue biomarker analysis embedded in a phase II clinical trial of cytarabine in castration-refractory prostate cancer to investigate prostate cancer biology

Abstract Background: Other than the androgen receptor, the TMPRSS2-ERG genomic aberrations in prostate cancer provide the first recent opportunity to target therapy in castration refractory prostate cancer (CRPC). We initiated a phase 2 clinical trial of cytarabine in docetaxel refractory CRPC on the basis of microarray, in vitro and case report evidence that cytarabine may be particularly effective in men harbouring abnormalities of the ERG oncogenes. Embedded in this clinical trial was the first use of blood mRNA levels of prostate cancer related genes as biomarkers of response and prognosis. Methods: Patients with docetaxel refractory progressive CRPC received intravenous cytarabine at doses between 1g/m2- 0.25 g/m2 q3 weekly. Responses were defined according to PCWG2C. 10 patients were enrolled between June 2007 and January 2010. TMPRSS2:ERG, PSA and PCA3 mRNA copies in whole blood collected with PAXgene tubes at the beginning of each cycle and at trial termination were quantified using transcription-mediated amplification assays. The prototype TMPRSS2:ERG assay detects the gene fusion isoform TMPRSS2 exon1 to ERG exon4. Results: No patients demonstrated a serum PSA response (PCWG2C). The average number of cycles administered was 2.6. Significant toxicities including grade 3-4 thrombocytopenia (2) and grade 3-4 neutropenia (3). These toxicities necessitated several dose reductions in the protocol, however most patients were removed from trial for serum PSA progression alone. PCA3 and PSA mRNAs were detectable in 8/10 and 9/10 cases, respectively; there was no correlation between serum PSA and PCA3 or PSA mRNA copy levels in blood. Testing for TMPRSS2:ERG in blood was able to predict the presence or absence of the TMPRSS2-ERG rearrangement in 9/10 cases when compared to 3 colour FISH carried out on baseline biopsies/ prostatectomies (2/10 positive for Exon 4:Exon 1 deletion). Conclusions: Cytarabine administation is ineffective in docetaxel refractory CRPC. Blood mRNA levels of prostate cancer genes reveal novel aspects of prostate cancer biology and have implications for the understanding of circulating tumour cells Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 290. doi:10.1158/1538-7445.AM2011-290

Utility of PCA3 urine assay in Japanese men undergoing prostate biopsy.

50 Background: Studies have shown the high specificity of PCA3 urine assay for biopsy detection of prostate cancer in European and American. We determined the diagnostic performance of PCA3 urine test in predicting prostate cancer in Japanese men. Methods: The study group included 261 men who underwent extended prostate biopsy at our institutions. 79 men of them had repeated biopsy. In all cases, race was Asian. Urine specimens were collected after digital rectal examination. On PROGENSA PCA3 assay, PCA3 score was calculated using PCA3 mRNA copy divided by PSA mRNA copy. PCA3 score was correlated with biopsy outcome and compared with serum PSA and free-total PSA. Results: 257 urine samples collected were successfully analyzed (i.e., the informative specimen rate was 98%). Biopsy revealed prostate cancer in 104 men (40%). There was no significant relationship between serum PSA and PCA3 score. PCA3 score in prostate cancer was significantly higher than that in negative (median 54.5 vs 17.0, p&lt;0.001). The probability of positive biopsy increased as PCA3 score increased. Using a PCA3 cutoff value of 35.0, sensitivity, specificity and diagnostic accuracy were 65%, 75% and 71%, respectively (p&lt;0.01). Area under the curve (AUC) for PCA3 score was 0.757 and AUC for serum PSA was 0.599 in all men. There was a significant difference in AUC between PCA3 score and serum PSA (p&lt;0.001). In men with serum PSA 4 to 10ng/ml, AUCs of PCA3 score, free-total PSA, and serum PSA were 0.759, 0.681, and 0.542, respectively. Conclusions: The specificity of PCA3 urine assay was high for detection of prostate cancer in Japanese men. PCA3 urine assay could improve the prediction for prostate cancer and help better decision making for prostate biopsy. No significant financial relationships to disclose.