Glyphosate Exposure Induces Cytotoxicity, Mitochondrial Dysfunction and Activation of ERα and ERß Estrogen Receptors in Human Prostate PNT1A Cells.

Glyphosate, the active ingredient of several broad-spectrum herbicides, is widely used throughout the world, although many adverse effects are known. Among these, it has been recognized as an endocrine disruptor. This work aimed to test the effects and potential endocrine disrupting action of glyphosate on PNT1A human prostate cells, an immortalized non-tumor epithelial cell line, possessing both ERα and ERß estrogen receptors. The results showed that glyphosate induces cytotoxicity, mitochondrial dysfunction, and rapid activation of ERα and ERß via nuclear translocation. Molecular analysis indicated a possible involvement of apoptosis in glyphosate-induced cytotoxicology. The apoptotic process could be attributed to alterations in mitochondrial metabolism; therefore, the main parameters of mitochondrial functionality were investigated using the Seahorse analyzer. Impaired mitochondrial function was observed in glyphosate-treated cells, with reductions in ATP production, spare respiratory capacity, and proton leakage, along with increased efficiency of mitochondrial coupling. Finally, the results of immunofluorescence analysis demonstrated that glyphosate acts as an estrogen disruptor determining the nuclear translocation of both ERs. Nuclear translocation occurred independent of dose, faster than the specific hormone, and persisted throughout treatment. In conclusion, the results collected show that in non-tumor prostate cells glyphosate can cause cell death and acts as a xenoestrogen, activating estrogen receptors. The consequent alteration of hormonal functions can have negative effects on the reproductive health of exposed animals, compromising their fertility.

Unraveling the Metabolic Alterations Induced by Zika Infection in Prostate Epithelial (PNT1a) and Adenocarcinoma (PC-3) Cell Lines.

The outbreak of Zika virus infection in 2016 led to the identification of its presence in several types of biofluids, including semen. Later discoveries associated Zika infection with sexual transmission and persistent replication in cells of the male reproductive tract. Prostate epithelial and carcinoma cells are favorable to virus replication, with studies pointing to transcriptomics alterations of immune and inflammation genes upon persistence. However, metabolome alterations promoted by the Zika virus in prostate cells are unknown. Given its chronic effects and oncolytic potential, we aim to investigate the metabolic alterations induced by the Zika virus in prostate epithelial (PNT1a) and adenocarcinoma (PC-3) cells using an untargeted metabolomics approach and high-resolution mass spectrometry. PNT1a cells were viable up to 15 days post ZIKV infection, in contrast to its antiproliferative effect in the PC-3 cell lineage. Remarkable alterations in the PNT1a cell metabolism were observed upon infection, especially regarding glycerolipids, fatty acids, and acylcarnitines, which could be related to viral cellular resource exploitation, in addition to the over-time increase in oxidative stress metabolites associated with carcinogenesis. The upregulation of FA20:5 at 5 dpi in PC-3 cells corroborates the antiproliferative effect observed since this metabolite was previously reported to induce PC-3 cell death. Overall, Zika virus promotes extensive lipid alterations on both PNT1a and PC-3 cells, promoting different outcomes based on the cellular metabolic state.

Bisphenol A modulates proliferation, apoptosis, and wound healing process of normal prostate cells: Involvement of G2/M-phase cell cycle arrest, MAPK signaling, and transcription factor-mediated MMP regulation.

Bisphenol A (BPA) is commonly used to produce epoxy resins and polycarbonate plastics. BPA is an endocrine-disrupting chemical that is leaked from the polymer and absorbed into the body to disrupt the endocrine system. Although BPA may cause cytotoxicity in the prostate, a hormone-dependent reproductive organ, its underlying mechanism has not yet been elucidated. Here, we investigated the effects of BPA on cell proliferation, apoptosis, and the wound healing process using prostate epithelial cells (RWPE-1) and stromal cells (WPMY-1). Observations revealed that BPA induced G2/M cell cycle arrest in both cell types through the ATM-CHK1/CHK2-CDC25c-CDC2 signaling pathway, and the IC50 values were estimated to be 150 µM. Furthermore, BPA was found to induce caspase-dependent apoptosis through initiator (caspase-8 and -9) and executioner (caspase-3 and -7) caspase cascades. In addition, BPA interfered with the wound healing process through inhibition of MMP-2 and - 9 expression, accompanied by reductions in the binding activities of AP-1 as well as NF-κB motifs. Phosphorylation of MAPKs was associated with the BPA-mediated toxicity of prostate cells. These results suggest that BPA exhibits prostate toxicity by inhibiting cell proliferation, inducing apoptosis, and interfering with the wound healing process. Our study provided new insights into the precise molecular mechanisms of BPA-induced toxicity in human prostate cells.

Pyruvate-lactate exchange and glucose uptake in human prostate cancer cell models. A study in xenografts and suspensions by hyperpolarized [1-13 C]pyruvate MRS and [18 F]FDG-PET.

Reprogramming of energy metabolism in the development of prostate cancer can be exploited for a better diagnosis and treatment of the disease. The goal of this study was to determine whether differences in glucose and pyruvate metabolism of human prostate cancer cells with dissimilar aggressivenesses can be detected using hyperpolarized [1-13 C]pyruvate MRS and [18 F]FDG-PET imaging, and to evaluate whether these measures correlate. For this purpose, we compared murine xenografts of human prostate cancer LNCaP cells with those of more aggressive PC3 cells. [1-13 C]pyruvate was hyperpolarized by dissolution dynamic nuclear polarization (dDNP) and [1-13 C]pyruvate to lactate conversion was followed by 13 C MRS. Subsequently [18 F]FDG uptake was investigated by static and dynamic PET measurements. Standard uptake values (SUVs) for [18 F]FDG were significantly higher for xenografts of PC3 compared with those of LNCaP. However, we did not observe a difference in the average apparent rate constant kpl of 13 C label exchange from pyruvate to lactate between the tumor variants. A significant negative correlation was found between SUVs from [18 F]FDG PET measurements and kpl values for the xenografts of both tumor types. The kpl rate constant may be influenced by various factors, and studies with a range of prostate cancer cells in suspension suggest that LDH inhibition by pyruvate may be one of these. Our results indicate that glucose and pyruvate metabolism in the prostate cancer cell models differs from that in other tumor models and that [18 F]FDG-PET can serve as a valuable complementary tool in dDNP studies of aggressive prostate cancer with [1-13 C]pyruvate.

Potential role of CFTR in bisphenol A-induced malignant transformation of prostate cells via mitochondrial apoptosis.

Bisphenol A (BPA) is an environmental endocrine disruptor and a risk factor for prostate cancer. The cystic fibrosis transmembrane conductance regulator (CFTR) is proposed to be a prostate cancer suppressor in some recent researches. However, the potential role and mechanism of CFTR in BPA-induced prostate cancer cells has not been well identified. In this study, BPA decreased the viability of human normal prostate RWPE-1 cells detected with a CCK-8 kit. The capacity of the cell line on soft agar colony formation, wound healing, and transwell invasion indicated malignant transformation induced by BPA. Western blot analysis demonstrated that the levels of CFTR and Bcl-2 decreased, whereas Bax level increased, and ELISA detection showed a decreased ATP level in BPA-exposed cells. Cell apoptosis was analyzed with Annexin V-FITC Detection Kit by flow cytometry. However, no significant difference was observed in cell viability and apoptosis rates compared to normal RWPE-1 cells. Our research revealed a potential role of CFTR in BPA-induced malignant transformation via mitochondrial apoptosis of normal prostate cells.

Hyaluronan-Metal Gold Nanoparticle Hybrids for Targeted Tumor Cell Therapy.

In this study, a novel multifunctional nanoplatform based on core-shell nanoparticles of spherical gold nanoparticles (AuNPs) capped with low and high molecular weight (200 and 700 kDa) hyaluronic acid (HA), was assembled via a green, one-pot redox synthesis method at room temperature. A multitechnique characterization approach by UV-visible spectroscopy, dynamic light scattering and atomic force microscopy pointed to the effective 'surface decoration' of the gold nanoparticles by HA, resulting in different grafting densities of the biopolymer chains at the surface of the metal nanoparticle, which in turn affected the physicochemical properties of the nanoparticles. Specifically, the spectral features of the gold plasmonic peak (and the related calculated optical size), the hydrodynamic diameter and the nanoparticle stability were found to depend on the molecular weight of the HA. The CD44-targeting capability of HA-functionalized gold nanoparticles was tested in terms of antibacterial activity and cytotoxicity. An enhanced inhibitory activity against both Gram-negative Escherichia coli and Gram-positive Staphylococcus aureus was found, with a HA molecular weight (MW)-dependent trend for the HA-capped AuNPs compared to the bare, glucose-capped AuNPs. Cell viability assays performed on two CD44-positive cell models, namely normal human umbilical vein endothelial (HUVEC) and prostate tumor (PC-3) cells, in comparison with neuroblastoma cells (SH-SY5Y), which do not express the CD44 receptor, demonstrated an increased cytotoxicity in neuroblastoma compared to prostate cancer cells upon the cellular treatments by HA-AuNP compared to the bare AuNP, but a receptor-dependent perturbation effect on cytoskeleton actin and lysosomal organelles, as detected by confocal microscopy. These results highlighted the promising potentialities of the HA-decorated gold nanoparticles for selective cytotoxicity in cancer therapy. Confocal microscopy imaging of the two human tumor cell models demonstrated a membrane-confined uptake of HA-capped AuNP in the cancer cells that express CD44 receptors and the different perturbation effects related to molecular weight of HA wrapping the metallic core of the plasmonic nanoparticles on cellular organelles and membrane mobility.

Comparative Cytotoxicity induced by Zinc Oxide Nanoparticles in Human Prostate Cells.

Despite increasing biomedical applications of zinc oxide nanoparticles (ZnO NPs), there is a lack of information concerning the biological effects of ZnO NPs on human cells. The purpose of this study was to assess the comparative cytotoxicity in human prostate cells (PC-3 and RWPE-1) exposure to 20-nm ZnO NPs. Exposure to concentrations from 0 to 50 µg/mL of ZnO NPs reduced cell viability of PC-3 cells in a dose- and time-dependent manner; whereas it did not affect RWPE-1 cells. A dose-dependent increase in LDH leakage and intracellular reactive oxygen species was observed in PC-3 cells but not in RWPE-1 cells exposure to ZnO NPs at concentrations of 8 ˜ 50 µg/mL for 24 h (P < 0.05). That the percentage of apoptotic cells increased significantly was observed in PC-3 cells induced by ZnO NPs at 10 µg/mL exposure for 8 h. Our results showed that ZnO NPs induced in vitro preferential cytotoxicity in the human prostate cancer cells. We indicated that the different cytotoxicity of ZnO NPs is likely due to the different cell-nanoparticle interaction and response behavior rather than to hydrodynamic sizes of particles. It is suggested that ZnO NPs are expected to find a very promising targeting therapeutic application for human prostate cancer.

Prediction model for early biochemical recurrence after radical prostatectomy based on the Cancer of the Prostate Risk Assessment score and the presence of secondary circulating prostate cells.

OBJECTIVE: To establish a prediction model for early biochemical recurrence based on the Cancer of the Prostate Risk Assessment (CAPRA) score and the presence of secondary circulating prostate cells (CPCs). PATIENTS AND METHODS: We conducted a prospective single-centre study of men who underwent radical prostatectomy as monotherapy for prostate cancer. Clinicopathological findings were used to calculate the CAPRA score. At 90 days after surgery, blood was taken for CPC detection, mononuclear cells were obtained using differential gel centrifugation, and CPCs were identified using immunocytochemistry. A CPC was defined as a cell expressing prostate-specific antigen (PSA) but not CD45. The CPC test results were defined as positive or negative. Patients were followed up for up to 5 years and biochemical recurrence was defined as a PSA level >0.2 ng/mL. The validity of the CAPRA score was calibrated using partial validation, and Cox proportional hazard regression to build three models: a CAPRA score model, a CPC model and a CAPRA/CPC combined model. RESULTS: A total of 321 men, with a mean age of 65.5 years, participated in the study. After 5 years of follow-up the biochemical recurrence-free survival rate was 98.55%. For the model that included CAPRA score there was a hazard ratio (HR) of 7.66, for the CPC model there was an HR of 34.52 and for the combined model there were HRs of 2.60 for CAPRA score and 22.5 for CPC. Using the combined model, 23% of men changed from the low-risk to the high-risk category, or vice versa. CONCLUSION: The incorporation of CPC detection significantly improved the model's discriminative ability in establishing the probability of biochemical recurrence; patients in the high-risk group according to CAPRA score who are negative for CPCs have a much better prognosis. The addition of CPC detection gives clinically significant information to aid the decision on who may be eligible for adjuvant therapy.

Convergent Effects of Resveratrol and PYK2 on Prostate Cells.

Resveratrol, a dietary polyphenol, is under consideration as chemopreventive and chemotherapeutic agent for several diseases, including cancer. However, its mechanisms of action and its effects on non-tumor cells, fundamental to understand its real efficacy as chemopreventive agent, remain largely unknown. Proline-rich tyrosine kinase 2 (PYK2), a non-receptor tyrosine kinase acting as signaling mediator of different stimuli, behaves as tumor-suppressor in prostate. Since, PYK2 and RSV share several fields of interaction, including oxidative stress, we have investigated their functional relationship in human non-transformed prostate EPN cells and in their tumor-prone counterpart EPN-PKM, expressing a PYK2 dead-kinase mutant. We show that RSV has a strong biological activity in both cell lines, decreasing ROS production, inducing morphological changes and reversible growth arrest, and activating autophagy but not apoptosis. Interestingly, the PYK2 mutant increases basal ROS and autophagy levels, and modulates the intensity of RSV effects. In particular, the anti-oxidant effect of RSV is more potent in EPN than in EPN-PKM, whereas its anti-proliferative and pro-autophagic effects are more significant in EPN-PKM. Consistently, PYK2 depletion by RNAi replicates the effects of the PKM mutant. Taken together, our results reveal that PYK2 and RSV act on common cellular pathways and suggest that RSV effects on prostate cells may depend on mutational-state or expression levels of PYK2 that emerges as a possible mediator of RSV mechanisms of action. Moreover, the observation that resveratrol effects are reversible and not associated to apoptosis in tumor-prone EPN-PKM cells suggests caution for its use in humans.

Potential impacts of bisphenols on prostate cells: An overview of cytotoxicity, proliferation, oxidative stress, apoptosis, and ER-stress response activation.

This study aimed to evaluate the toxic effects of Bisphenol A (BPA), Bisphenol F (BPF) and Bisphenol S (BPS) on PNT1A and PC-3 cells, focusing on their effects on endoplasmic reticulum (ER) stress and related pathways. PNT1A and PC-3 were treated with BPA, BPF and BPS at concentrations of 0.1, 1 and 10 µM for 48 h cytotoxicity, BrdU cell proliferation, ROS generation, apoptosis detection, gene expression analysis and Western blot analysis were performed. BPA induced proliferation and late apoptosis in PNT1A cells, whereas it induced both late apoptosis and early apoptosis in PC-3 cells. BPF and BPS induced late apoptosis in PC-3 cells. Increased ROS levels were observed in PNT1A cells exposed to 1-10 µM BPA. BPA, BPF and BPS increased the expression levels of ER stress-related genes in PNT1A cells. Furthermore, exposure to BPA increased the expression of ER stress-related CHOP/DDIT3 protein in PNT1A cells. These findings highlight the potential health risks associated with BPA, BPF and BPS exposure and emphasize the importance of investigating the underlying mechanisms by which these chemicals may affect human health. Further research is required to comprehensively understand the role of ER stress pathways in cellular responses to these substances.

In Vitro Modelling of Chlamydia trachomatis Infection in the Etiopathogenesis of Male Infertility and Reactive Arthritis.

Chlamydia trachomatis is an obligate, intracellular bacterium responsible for a range of diseases of public health importance, since C. trachomatis infection is often asymptomatic and, hence, untreated, leading to chronic complications, including prostatitis, infertility, and reactive arthritis. The ample spectrum of diseases caused by C. trachomatis infection is reflected in its ability to infect and multiply within a wide range of different cell types. Cervical epithelial cells, to date, have been the most studied cellular infection model, highlighting the peculiar features of the host-cell inflammatory and immune responses to the infection. Herein, we provide the up-to-date evidence on the interaction between C. trachomatis and human prostate epithelial, Sertoli and synovial cells.

RBE variation in prostate carcinoma cells in active scanning proton beams: In-vitro measurements in comparison with phenomenological models.

PURPOSE: In-vitro radiobiological studies are essential for modelling the relative biological effectiveness (RBE) in proton therapy. The purpose of this study was to experimentally determine the RBE values in proton beams along the beam path for human prostate carcinoma cells (Du-145). RBE-dose and RBE-LETd (dose-averaged linear energy transfer) dependencies were investigated and three phenomenological RBE models, i.e. McNamara, Rørvik and Wilkens were benchmarked for this cell line. METHODS: Cells were placed at multiple positions along the beam path, employing an in-house developed solid phantom. The experimental setup reflected the clinical prostate treatment scenario in terms of field size, depth, and required proton energies (127.2-180.1 MeV) and the physical doses from 0.5 to 6 Gy were delivered. The reference irradiation was performed with 200 kV X-ray beams. Respective (α/ß) values were determined using the linear quadratic model and LETd was derived from the treatment planning system at the exact location of cells. RESULTS AND CONCLUSION: Independent of the cell survival level, all experimental RBE values were consistently higher in the target than the generic clinical RBE value of 1.1; with the lowest RBE value of 1.28 obtained at the beginning of the SOBP. A systematic RBE decrease with increasing dose was observed for the investigated dose range. The RBE values from all three applied models were considerably smaller than the experimental values. A clear increase of experimental RBE values with LETd parameter suggests that proton LET must be taken into consideration for this low (α/ß) tissue.

Detecting acid phosphatase enzymatic activity with phenol as a chemical exchange saturation transfer magnetic resonance imaging contrast agent (PhenolCEST MRI).

Phenol contains an exchangeable hydroxyl proton resonant at 4.8 ppm from the resonance frequency of water in the 1H nuclear magnetic resonance (1H NMR) spectrum, enabling itself to be detected at sub-mM concentration by either chemical exchange saturation transfer magnetic resonance imaging (CEST MRI) or exchange-based T2 relaxation enhancement (T2ex) effect under acidic and basic conditions, respectively. We recently investigated the T2ex effects of phenol and its derivatives, but the CEST characteristics of phenols are unknown in detail, and no study on using the natural CEST MRI effects of phenol for detecting enzymatic activity has been conducted. Herein, on the basis of the inherent CEST MR property of phenol, namely phenolCEST, we developed the first MRI approach to detect acid phosphatase (AcP) enzymatic activity. Upon the activity of AcP at pH = 5.0, non-CEST-detectable enzyme substrate phenyl phosphate was converted to CEST-detectable phenol, providing a simple way to quantify AcP activity directly without the need for a second signalling probe. We showed the application of this phenolCEST biosensor for measuring AcP activity in both enzyme solutions and cell lysates of prostate cells. This work opens a door for the utilization of phenolCEST MRI technique in sensor design and development.

Predictive Value of Neutrophil to Lymphocyte Ratio in the Diagnosis of Significant Prostate Cancer at Initial Biopsy: A Comparison with Free Percent Prostate Specific Antigen, Prostate Specific Antigen Density and Primary Circulating Prostate Cells.

INTRODUCTION: An elevated serum PSA is the only biomarker routinely used in screening for prostate cancer to indicate a prostate biopsy. However, it is not specific for prostate cancer and the neutrophil/lymphocyte ratio has been suggested as an alternative. We present a prospective study of men with an elevated PSA and compare the neutrophil/lymphocyte ratio, free percent PSA, PSA density and the presence of circulating prostate cells to detect clinically significant prostate cancer at first biopsy. PATIENTS AND METHODS: Prospective study of consecutive men with a PSA 4-10 ng/ml referred for initial prostate biopsy, the results were compared with the neutrophil/lymphocyte ratio, free percent PSA and PSA density. Circulating prostate cells (CPCs) were detected using immunocytochemistry. The blood sample was taken immediately before the prostate biopsy. RESULTS: 1,223 men participated, 38% (467) of whom had prostate cancer detected, of these 322 were clinically significant. The area under the curves were for neutrophil/lymphocyte ratio, free percent PSA, PSA density and CPC detection were 0.570, 0.785, 0,620 and 0.844 respectively. Sensitivity/specificity were 0.388/0.685, 0.419/0.897, 0.598/0.624 and 0.966/0.786 respectively. The neutrophil/lymphocyte ratio did not differentiate between benign and malignant disease. CONCLUSIONS: The neutrophil/lymphocyte ratio did not discriminate between benign and malignant prostatic disease in patients with a PSA between 4-10ng/ml.

[Primary circulating prostate cells, age and prostatic volumen predict the presence of prostate cancer in men with indication of a second prostate biopsy.] / Las células prostáticas circulantes primarias, edad y volumen prostático predicen el cáncer prostático en sujetos con indicación de una segunda biopsia prostática. Primary circulating prostate cells, age and prostatic volumen predict the presence of prostate cancer in men with indication of a second prostate biopsy.

INTRODUCTION: The limitations of serumPSA as a screening test to detect prostate cancer remainproblematic, especially after an initial negative prostatebiopsy. Detection of primary circulating prostate cells(CPCs) has been reported to be useful in the detectionof prostate cancer in men with a serum PSA>4.0ng/ml.We present a prospective study comparing the detectionof CPCs, total PSA, percent free PSA, digital rectal examination(DRE) and prostate volumen (PV) to establisha predictive model for the detection of prostate cancerin men with an indication for a second prostate biopsy. OBJECTIVE: To establish a predictive model for the detectionof prostate cancer using the number of CPCs detectedper sample, DRE, age, total serum PSA, percentfree PSA and PV in men with an indication for a secondprostate biopsy. METHODS AND PATIENTS: A prospective, observationalstudy carried out in the Hospital de Carabineros deChile, between 2006 and 2014 including 199 menundergoing a second prostate biopsy. The variables,number of CPCs detected (nCPC), DRE, age, PSA, percentfree PSA and PV were registered for each patientand based on these findings and comparing them withthe results of the prostate biopsy a multivariate logistic regressionanalysis incorporating forward predictors. Themodel was evaluated for co-lineal tendency, reliabilityand error specificity, and analyzed using non-parametricreceiver operating characteristics and area under thecurve decision for the combined model and for eachvariable separately. RESULTS: The single variable nCPC had a superior predictivevalue with an area under the curve of 0.89 (95%CI 0.83-0.94). The final model incorporated nCPC (OR:2.03 95% CI 1.63-2.53) age (OR: 1.1 95% CI 1.04-1.17) and PV (OR: 0.96 95% CI 0.93-0.99) with anarea under the curve for the combined model of 0.92(95% CI 0.88-0.97). The combined model performedbetter than the variables used alone. CONCLUSIONS: The model incorporating nCPC, ageand PV had a greater diagnostic yield for the predictingprostate cancer at second biopsy.

INTRODUCCIÓN: Las limitaciones del antígeno prostático específico (PSA) total como examen de pesquisa de cáncer prostático sigue problemático, especialmente después de una biopsia prostática inicial negativa. El uso de la detección de células prostáticascirculantes primarias (CPCs) ha sido reportado de ser útil en la detección de cáncer prostático en hombres con un APE >4,0 ng/ml.OBJETIVO: Presentamos un estudio prospectivo utilizando la detección de CPCs, el PSA, el porcentaje libre del PSA, el tacto rectal (TR) y el volumen prostático (VP) para establecer un modelo predictivo para la detección de cáncer prostático en pacientes con indicación de una segunda biopsia prostática.MÉTODOS Y PACIENTES: En Hospital de Carabineros de Chile entre 2006 y 2014, se realiza un estudio observacional, consignando para 199 sujetos, en forma previa a segunda biopsia de próstata, lo siguiente: nCPC, TR, edad, PSA, porcentaje libre de PSA y VP. Apartir de estas variables y en relación a resultado de neoplasia en estudio histológico de próstata, se efectúa una regresión logística multivariante con incorporación "forward" de predictores. En el modelo seleccionado se efectúa una evaluación del ajuste, co-linealidad predictores y especificación del error. Además se realizaanálisis de "Receiver Operating Characteristic" (ROC) no paramétrico y análisis de curva de decisión (ACD) para el modelo obtenido y también para los predictores en forma separada.RESULTADOS: Como variable separado el nCPC tuvo un rendimiento superior con un área ROC de 0,89 (CI 95% 0,83 a 0,94). El modelo final incorpora nCPC (OR: 2,03; CI95%: 1,63 a 2,53), edad (OR: 1,1; CI95%: 1,04 a 1,17) y VP (OR: 0,96; CI95%: 0,93 a 0,99) con un rendimiento en área ROC (0,92; CI95%: 0,88 a 0,97) y ACD mayor al resto de las variables por separado.CONCLUSIÓN: El modelo que incorpora el NCPC, edad y VP, presenta un rendimiento mayor en la predicción de la presencia de malignidad, en sujetos que realizan una segunda biopsia prostática.

[Circulating prostate cells and bone marrow micrometastasis are determinant in risk and time to biochemical progression in localized prostate cancer.] / Células prostáticas circulantes y micro-metástasis en médula ósea determinantes en el riesgo y en el tiempo de progresión bioquímica en cáncer prostático localizado.

INTRODUCTION: Minimal residual disease (MRD) is that which remains after curative therapy for prostate cancer. It has the potential for growth and later cause metastasis. After radical prostatectomy, the detection of circulating prostate cells (CPCs) and bone marrow micro-metastasis could represent different types of MRD. We proposed to determine the biochemical failure free survival rates, the time to biochemical failure after 10 years of follow-up and the presence of CPCs and micro-metastasis in patients treated with RP for pathologically organ confined prostate cancer. METHODS AND PATIENTS: One month after RP monotherapy for prostate cancer, blood and bone marrow samples were taken to detect CPCs and micro-metastasis. Men were classified as: group A (CPC negative and micro-metastasis negative), group B (CPC negative and micro-metastasis positive), group C (CPC positive and micro-metastasis negative), and group D (CPC positive and micro-metastasis positive). All subjects were followed with serial total PSA levels, recording the time at which failure occurred defined as a serum PSA > 0.2ng/ ml on two separate occasions. After ten years of follow- up for each group Kaplan-Meier survival curves were determined and using an adjusted flexible parametric model (FP), the Restricted Mean Survival Times for groups A, B, C and D were calculated. RESULTS: 191 men participated, 10-year biochemical failure survival rates were; group A (N=114) with a Kaplan-Meier of 98.7%; group B (N=39) 65.1%; group C (N=12) 10.4% and in group D (N=28) 12.8%. The Restricted Mean Survival Times (years) were group A: 9.95; group B: 9.45, group C: 5.11 and group D: 6.18 (p-value <0.001 between groups: A versus C, Aversus D, B versus C and B versus D). Frequency and time to failure was dependent on the type of MRD, those men CPC positive had a significantly higher failure rate and early failure. Those men only micro-metastasis positive had lower failure rate and late failure when compared with men negative for MRD. CONCLUSIONS: CPC positive men have a more aggressive disease with increased early failure; those men who are only positive for micro-metastasis are at risk for late or delayed failure. These two forms of measuring MRD represent different stages in the disease progression and may be used to guide clinical treatment decisions before increases in PSA levels.

INTRODUCCIÓN: La enfermedad mínima residual es aquella que persiste tras un tratamiento curativo. Tiene el potencial para crecer y causar metástasis. Tras una prostatectomía radical, la detección de células prostáticas circulantes (CPCs) y micrometástasis en médula ósea pueden representar diferentes tipos de la enfermedad mínima residual. Proponemos determinar la tasa de supervivencia libre de recidiva bioquímica, el tiempo hasta la recidiva bioquímica y la presencia de CPCs y micrometástasis en pacientes sometidos a prostatectomia por cáncer prostático localizado.MÉTODOS Y PACIENTES: Se tomaron muestras de sangre y médula ósea un mes después de la cirugía, para detectar CPCs y micro-metástasis. Los sujetos fueron clasificados en 4 grupos: Grupo A (CPCs negativo, micro-metástasis negativo), Grupo B (CPCs negativo, micro-metástasis positivo), Grupo C (CPCs positivo, micro-metástasis negativo), Grupo D (CPCs positivo, micro-metástasis positivo). Los sujetos se siguieron con PSA sérico; la recidiva bioquímica se definió como un PSA > 0,2ng/ml. Tras 10 años de seguimiento se determinó la curva se sobrevida de Kaplan-Meier y mediante un modelo paramétrico flexible ajustado, se calculó el tiempo de supervivencia medio restringido para todos los grupos.RESULTADOS: Participaron 191 hombres. La tasa de supervivencia a 10 años (KaplanMeier) en Grupo A (N=114) fue 98,7%, Grupo B (N=39) 65,1%, Grupo C (N=12) 10,4% y Grupo D (N=28) 12,8%. El tiempo de supervivencia medio restringido (años) fue; GrupoA 9,95, Grupo B 9,45, Grupo C 5,11 y Grupo D fue 6,18 (valor p<0,001 entre A versus C, A versus D,  B versus C y B versus D). La frecuencia y el tiempo hasta la recidiva fue dependiente del tipo de enfermedad mínima residual, sujetos CPC positivos tuvieron una tasa de recidiva significativamente mayor y más temprana. Sujetos que solo presentaron micro-metástasis tuvieron menor tasa de recidiva y más tardía que aquellos sin enfermedad mínima residual.CONCLUSIONES: Hombres CPC positivas presentaron una enfermedad más agresiva con recidiva temprana. Hombres con solo micro-metástasis tienen riesgo de recidiva tardía. Estas dos formas de medir la enfermedad mínima residual representan diferentes entidades clínicas y podría ser utilizada como una guía para la toma de decisiones clínicas previo al aumento del PSA sérico.

The presence of secondary circulating prostate tumour cells determines the risk of biochemical relapse for patients with low- and intermediate-risk prostate cancer who are treated only with external radiotherapy.

INTRODUCTION: The classification of patients with prostate cancer is used to determine treatments based on risk factors. The presence of secondary circulating prostate tumour cells (CPCs) detected in peripheral blood after a curative treatment has been associated with a worse prognosis. We present a prospective study of CPC detection post radiotherapy and the oncological results. PATIENTS AND METHODS: All of the patients classified as low and intermediate risk that were treated with radiotherapy were included. Three months after finishing treatment, an 8-ml blood sample was taken to detect CPCs. Mononuclear cells were obtained using gel centrifugation, and CPCs were identified using immunocytochemistry with anti-prostate-specific antigen. Patients were classified as low-risk CPC positive or negative and intermediate-risk CPC positive or negative. The biochemical relapse-free survival analysis was determined based on a follow-up of up to 15 years using the Kaplan-Meier and Cox regression models. Biochemical failure was defined according to the Pheonix II criteria. RESULTS: Of 241 patients, 181 (75.1%) were classified as low risk and 60 (24.9%) as intermediate risk. Biochemical failure was observed in 27.1% (49/181) of the low-risk prostate cancer participants and in 53.3% (32/60) of intermediate-risk participants after 15 years of follow-up. 20.4% (37/181) of the low-risk cancer participants had detectable CPCs in comparison with 43.3% (26/60) of the intermediate-risk cancer participants (p < 0.001 overall risk 2.98, confidence interval (CI) 95% 1.59-5.56; relative risk 2.12, CI 95% 1.41-3.19). Positive CPC patients had a worse prognosis, and a shorter time period until biochemical relapse, regardless of risk group. The biochemical relapse-free survival curves show that intermediate-risk participants who were CPC negative had a higher survival rate and slower disease progression than those participants who were low risk but CPC positive. CONCLUSIONS: CPC detection is a risk factor for biochemical relapse and could be useful in identifying patients that will need additional treatment.

Combining the Prostate Cancer Risk Index (PRIX) with the Presence of Secondary Circulating Prostate Cells to Predict the Risk of Biochemical Failure after Radical Prostatectomy for Prostate Cancer

Introduction: The use of pre- and post-surgery variables has been used to create nomograms in order to identify patients at high risk of treatment failure. The PRIX nomogram is one such device; we compare the PRIX nomogram with the presence of secondary circulating prostate cells to predict those men who will undergo treatment failure. Methods and Patients: Men who underwent radical prostatectomy for prostate cancer entered the study. The PRIX score was calculated from the total serum PSA pre-surgery, the biopsy Gleason score and clinical stage. Circulating prostate cells were detected from venous blood one month after surgery, using differential gel centrifugation and standard immunocytochemistry with anti-PSA. A test was considered positive when 1 CPC/blood sample was detected. Patients were followed up for five years and biochemical failure was defined as a serum PSA >0.2ng/ml. Kaplan-Meier and Cox proportional models were used to calculate survival curves. Results: 321 men participated, of whom 131 (40.8%) underwent biochemical failure within 5 years. A higher PRIX score was associated with increased failure risk, as was the presence of CPCs. The predictive power of CPCs was significantly higher than the PRIX score. Combining the two methods, for equal PRIX scores, scores but CPC positive had a worse biochemical failure free survival than men with high PRIX scores but CPC negative. For men with PRIX scores of ≥4 the use of CPC detection did not aid in the clinical decision making process. For those with PRIX scores of 0 and 1, CPC detection identified men with a high risk of treatment failure. Conclusions: The combined PRIX/CPC score improved the predictive values of men at high risk of biochemical failure. Both are simple systems that could be incorporated in a general hospital. Further multicenter studies are warranted to confirm these results.

Minimum Residual Disease in Patients Post Radical Prostatectomy for Prostate Cancer: Theoretical Considerations, Clinical Implications and Treatment Outcome

Introduction: Minimal residual disease (MRD) remaining after curative therapy for prostate cancer has the potential for growth and can result in metastasis. Circulating prostate cells (CPCs) and bone marrow micro-metastasis (mM) could represent different types of MRD. We here determined; biochemical failure free survival rates; time to BF after 10 years follow-up; and the presence of CPCs and mM in patients treated with radical prostatectomy (RP) for prostate cancer. Methods and Patients: One month after RP, blood and bone marrow were sampled for assessment of CPCs and mM. Cases were classified as: group A, CPC negative and mM negative; group B, CPC negative and mM positive; Group C, CPC positive and mM negative; and Group D, CPC positive and mM positive. Subjects were followed with serial determination of PSA levels, recording the time at which BF occurred defined as a serum PSA >0.2ng/ml. After ten years of follow-up Kaplan-Meier survival curves were generated and the restricted mean survival time (RMST) for each group calculated. Results: A total of 321 men participated, 140 in group A with survival of 92.7% (86.3 to 96.2), 39 in group B with 55.8% (37.2 to 70.9); 54 in group C with 6.41% (1.19 to 18.21) and 88 in group D with 4.96%(1.64 to 11.13%. The RMST (in years) were: group A, 9.47 (9.24 to 9.69); group B, 9.23 (8.87 to 9.58); group C, 4.62 (4.46 to 4.77); and group D, 3.57 (3.52 and 3.63) (p-value<0.001 between groups: A versus C and D, B versus C and D). Conclusions: CPC positive men have more aggressive disease, with increased early failure; men who are only positive for mM are at greater risk of late failure. These two forms of MRD represent different clinical entities with respect to biochemical failure and could be used to guide clinical treatment decisions.

The presence of primary circulating prostate cells is associated with upgrading and upstaging in patients eligible for active surveillance.

Active surveillance (AS) is a considered treatment option for men with low or very low-risk prostate cancer. However, on repeat biopsy some 25% were upgraded and recommended for active treatment. We compare the presence or absence of primary circulating prostate cells (CPCs) with the clinical pathological findings after radical prostatectomy in men fulfilling the criteria for active surveillance and the risk of reclassification for active observation (AO). METHODS AND PATIENTS: A single centre observational study was done involving 102 men who fulfilled the Epstein criteria for AS and underwent radical prostatectomy as mono-therapy for prostate cancer. The patients were classified according to the presence or absence of CPCs detected immediately before the prostate biopsy. Mononuclear cells were obtained by differential gel centrifugation of 8 mL of venous blood and CPCs identified using immunocytochemistry with anti-PSA and anti-P504S. A positive CPC test was defined as at least 1 PSA (+), P504S (+) cell detected/blood sample. The surgical specimen was analysed for Gleason score and pathological stage. RESULTS: A total of 25 out of 102 (24.5%) men were upgraded based on the pathological findings of the surgical specimen. Among which 45 (44%) men were positive for CPCs. They were younger, 63.9 versus 68.1 years (p = 0.0148), had a lower frequency of pT2 or lower disease (64.4% versus 91.2% p <0.001), higher median Gleason scores (6 versus 5 p < 0.001) in both the biopsy and surgical specimens, and a higher frequency of upgrading 44% versus 9% (p < 0.001). CONCLUSIONS: In men fulfilling the criteria for AS, the presence of primary CPCs suggests a high risk for disease upgrade and therefore these men may not be ideal for observational therapy. Further studies with a larger population are warranted.