Peripheral Blood Of Prostate Cancer Patients Research Articles

Clinical assessment of TGFB1 and HP Relative Gene Expression in the Peripheral Blood of Prostate Cancer Patients.

This study aimed to assess the relative gene expression level of transforming growth factor-β1 (TGFB1) and haptoglobin (HP) in the peripheral blood of prostate cancer (PCa) patients and evaluate their diagnostic ability. A total of 125 participants were enrolled in the present study. Among them, 75 PCa patients, 25 benign prostatic hyperplasia (BPH) patients, and 25 healthy volunteers served as the control group. The relative TGFB1 and HP gene expression level was quantified using real-time polymerase chain reaction. Further, free and total PSA levels were determined using electrochemiluminescence assays. TGFB1 was significantly over-expressed, whereas HP was significantly downregulated in the peripheral blood of PCa patients compared to BPH and control groups (p-value ranges from 0.034 to <0.001). Moreover, the high expression level of TGFB1 was associated with an increased risk of PCa development with OR=1.412 (95%CI: 1.081-1.869, p= 0.012). TGFB1 and HP relative expression levels had lower diagnostic performance to differentiate PCa from normal and BPH individuals compared to PSA, however, the combination of the tested parameters improved the diagnostic efficacy. TGFB1 and HP relative expression have moderate diagnostic efficacy in discriminating patients with PCa from BPH and healthy subjects. Furthermore, over-expression of TGFB1 may contribute to the pathogenesis of PCa.

Frequencies of an Immunogenic HER-2/neu Epitope of CD8+ T Lymphocytes Predict Favorable Clinical Outcomes in Prostate Cancer

HER-2/neu is the human epidermal growth factor receptor 2, which is associated with the progression of prostate cancer (PCa). HER-2/neu-specific T cell immunity has been shown to predict immunologic and clinical responses in PCa patients treated with HER-2/neu peptide vaccines. However, its prognostic role in PCa patients receiving conventional treatment is unknown, and this was addressed in this study. The densities of CD8+ T cells specific for the HER-2/neu(780-788) peptide in the peripheral blood of PCa patients under standard treatments were correlated with TGF-β/IL-8 levels and clinical outcomes. We demonstrated that PCa patients with high frequencies of HER-2/neu(780-788)-specific CD8+ T lymphocytes had better progression-free survival (PFS) as compared with PCa patients with low frequencies. Increased frequencies of HER-2/neu(780-788)-specific CD8+ T lymphocytes were also associated with lower levels of TGF-β and IL-8. Our data provide the first evidence of the predictive role of HER-2/neu-specific T cell immunity in PCa.

Prostate Cancer Peripheral Blood NK Cells Show Enhanced CD9, CD49a, CXCR4, CXCL8, MMP-9 Production and Secrete Monocyte-Recruiting and Polarizing Factors.

Natural killer (NK) cells, effector lymphocytes of the innate immunity, have been shown to be altered in several cancers, both at tissue and peripheral levels. We have shown that in Non-Small Cell Lung Cancer (NSCLC) and colon cancer, tumour associated circulating NK (TA-NK) and tumour infiltrating NK (TI-NK) exhibit pro-angiogenic phenotype/functions. However, there is still a lack of knowledge concerning the phenotype of peripheral blood (PB) NK (pNK) cells in prostate cancer (PCa). Here, we phenotypically and functionally characterized pNK from PCa patients (PCa TA-NKs) and investigated their interactions with endothelial cells and monocytes/macrophages. NK cell subset distribution in PB of PCa patients was investigated, by multicolor flow cytometry, for surface antigens expression. Protein arrays were performed to characterize the secretome on FACS-sorted pNK cells. Conditioned media (CM) from FACS-sorted PCa pTA-NKs were used to determine their ability to induce pro-inflammatory/pro-angiogenic phenotype/functions in endothelial cells, monocytes, and macrophages. CM from three different PCa (PC-3, DU-145, LNCaP) cell lines, were used to assess their effects on human NK cell polarization in vitro, by multicolor flow cytometry. We found that PCa pTA-NKs acquire the CD56brightCD9+CD49a+CXCR4+ phenotype, increased the expression of markers of exhaustion (PD-1, TIM-3) and are impaired in their degranulation capabilities. Similar effects were observed on healthy donor-derived pNK cells, exposed to conditioned media of three different PCa cell lines, together with increased production of pro-inflammatory chemokines/chemokine receptors CXCR4, CXCL8, CXCL12, reduced production of TNFα, IFNγ and Granzyme-B. PCa TA-NKs released factors able to support inflammatory angiogenesis in an in vitro model and increased the expression of CXCL8, ICAM-1, and VCAM-1 mRNA in endothelial cells. Secretome analysis revealed the ability of PCa TA-NKs to release pro-inflammatory cytokines/chemokines involved in monocyte recruitment and M2-like polarization. Finally, CMs from PCa pTA-NKs recruit THP-1 and peripheral blood CD14+ monocyte and polarize THP-1 and peripheral blood CD14+ monocyte-derived macrophage towards M2-like/TAM macrophages. Our results show that PCa pTA-NKs acquire properties related to the pro-inflammatory angiogenesis in endothelial cells, recruit monocytes and polarize macrophage to an M2-like type phenotype. Our data provides a rationale for a potential use of pNK profiling in PCa patients.

Urinary spermine level as novel additional diagnostic marker of prostate cancer

Abstract Background prostate cancer (PC) is one of the most common male cancer in the world, and it is one of the leading causes of mortality and momentous public health impact in many developed countries. Nowadays, PSA (prostate-specific antigen) test is used as a famous serum test for screening of prostate cancer. However, PSA test isnt sensitive and specific enough, because its high levels do not always indicate the presence of a malignant process in the prostate, and low - about its absence. Some studies suggest to use concentration of spermine in the urine as a novel noninvasive marker for PC diagnostics. Methods Male patients age 51-79 with prostate cancer (I-IV stage) were enrolled. Diagnoses of all patients were established at the National Cancer Institute (Kyiv, Ukraine). Spermine concentration was determined in the morning urine samples of 38 patients with malignant tumors (I stage - 16%, II - 45%, III - 24%, IV - 15%) before treatment, 12 patients with prostate hyperplasia and 12 healthy men by ELISA kit. Results It was found that concentration of spermine in the urine in patients with prostate cancer was lower by 7-34 times in comparison with healthy men and lower by 5-13 times than in patients with prostate hyperplasia. The same results were obtained previously in peripheral blood of PC patients by the high pressure liquid chromatography (HPLC). Conclusion concentration of spermine in the urine shows potential to serve as a novel PC diagnostic non-invasive marker, which in turn can help to address the limited sensitivity and specificity problem of serum PSA test.

TUG1 promotes the development of prostate cancer by regulating RLIM.

The aim of this study was to investigate the specific role of Taurine up-regulated gene 1 (TUG1) in the development of prostate cancer (PCa), and to explore its underlying mechanism. The serum level of TUG1 in healthy subjects, benign prostatic hyperplasia (BPH) patients and PCa patients was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The correlation between TUG1 expression and clinical indexes of PCa patients was analyzed. TUG1 expression in PCa cells and human normal prostate cells was determined by qRT-PCR as well. Overexpression or knockdown of TUG1 was achieved by liposomal transfection. Subsequently, the regulatory effects of TUG1 on the proliferative and migratory capacities of DU145 cells were accessed by Cell Counting Kit-8 (CCK-8) assay, colony formation assay and transwell assay, respectively. An online software was used to predict whether RLIM could be regulated by TUG1, which was further verified by qRT-PCR. After RLIM knockdown in DU145 cells, the proliferative and migratory capacities were also determined. Finally, Western blot was conducted to determine relative protein expressions in the TGF-β1/Smad pathway after altering TUG1 expression in DU145 cells. TUG1 was highly expressed in serum samples of PCa patients when compared with healthy subjects and BPH patients. Besides, TUG1 expression in PCa patients with Gleason ≥ 7 was significantly higher than those with Gleason < 7. Meanwhile, TUG1 expression in PCa patients was remarkably higher than that of BPH patients at the PSA grey zone (4-10 ng/mg). ROC curves indicated that TUG1 might be a crucial hallmark to distinguish PCa patients from BPH patients and healthy subjects. The overexpression of TUG1 markedly promoted the proliferative and migratory capacities of DU145 cells. However, knockdown of TUG1 obtained the opposite results. QRT-PCR confirmed that TUG1 was positively correlated with RLIM at the mRNA level. RLIM knockdown significantly inhibited the proliferative and migratory capacities of DU145 cells. Furthermore, knockdown of TUG1 in DU145 cells markedly down-regulated TGF-β1 and p-Smad2, whereas up-regulated p-Smad7. TUG1 is highly expressed in peripheral blood of PCa patients, which can serve as a potential diagnostic marker for PCa. The overexpression of TUG1 promotes the proliferative and migratory capacities of PCa cells. Furthermore, TUG1 promotes the development of PCa by regulating RLIM through the TGF-β1/Smad pathway.

Heterogeneous PSMA expression on circulating tumor cells: a potential basis for stratification and monitoring of PSMA-directed therapies in prostate cancer.

The prostate specific membrane antigen (PSMA) is the only clinically validated marker for therapeutic decisions in prostate cancer (PC). Characterization of circulating tumor cells (CTCs) obtained from the peripheral blood of PC patients might provide an alternative to tissue biopsies called “liquid biopsy”. The aim of this study was to develop a reliable assay for the determination of PSMA on CTCs. PSMA expression was analyzed on tissue samples (cohort one, n = 75) and CTCs from metastatic PC patients (cohort two, n = 29). Specific signals for the expression of PSMA could be seen for different prostate cancer cell line cells (PC3, LaPC4, 22Rv1, and LNCaP) by Western blot, immunohistochemistry (IHC), immunocytochemistry (ICC), and FACS. PSMA expression was found to be significantly increased in patients with higher Gleason grade (p = 0.0011) and metastases in lymph nodes (p = 0.0000085) or bone (p = 0.0020) (cohort one). In cohort two, CTCs were detectable in 20 out of 29 samples (69 %, range from 1 - 1000 cells). Twelve out of 20 CTC-positive patients showed PSMA-positive CTCs (67 %, score 1+ to 3+). We found intra-patient heterogeneity regarding the PSMA status between CTCs and the corresponding primary tumors. The results of our study could help to address the question whether treatment decisions based on CTC PSMA profiling will lead to a measurable benefit in clinical outcome for prostate cancer patients in the near future.

130 CIRCULATING TUMOR CELLS IN PROSTATE CANCER PATIENTS: NOVEL IMMUNOMAGNETIC ENRICHMENT METHOD

You have accessJournal of UrologyProstate Cancer: Basic Research1 Apr 2011130 CIRCULATING TUMOR CELLS IN PROSTATE CANCER PATIENTS: NOVEL IMMUNOMAGNETIC ENRICHMENT METHOD Xiaoke Huang, Barry B. McGuire, Irene Ogden, Daniel C. O'Brien, Phillip R. Cooper, Jessica A. Banks, William J. Catalona, and Raymond C. Bergan Xiaoke HuangXiaoke Huang Chicago, IL More articles by this author , Barry B. McGuireBarry B. McGuire Chicago, IL More articles by this author , Irene OgdenIrene Ogden Chicago, IL More articles by this author , Daniel C. O'BrienDaniel C. O'Brien Chicago, IL More articles by this author , Phillip R. CooperPhillip R. Cooper Chicago, IL More articles by this author , Jessica A. BanksJessica A. Banks Chicago, IL More articles by this author , William J. CatalonaWilliam J. Catalona Chicago, IL More articles by this author , and Raymond C. BerganRaymond C. Bergan Chicago, IL More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2011.02.197AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Pharmacological prevention of metastases would improve quality of life for many prostate cancer (PCa) patients. To metastasize, cancer cells must invade through the basement membrane and the extracellular matrix of the organ's stromal compartment to intravasate into the bloodstream thereby becoming circulating tumor cells (CTCs). The number of CTCs correlates with the formation of distant metastases. Rates of PCa death are ∼10 fold lower among soy-consuming Southeast Asians than among non-soy consumers in the US. Genistein is a soy protein derivative that has been shown in vitro and in murine models to inhibit these early steps in the cascade. We implemented a Clinical Phase II Trial to determine whether genistein could decrease CTCs in men with prostate cancer (PCa). To measure this effect, we have developed a novel assay to detect and quantify CTCs in whole human blood. In addition to PSA, CTCs also express CK18. METHODS Patients were randomized to genistein treatment (2mg/kg of genistein per day three weeks prior to radical prostatectomy) or no treatment (placebo). A whole blood sample was obtained prior to surgery. After RNA extraction the expression of prostate specific antigen (PSA) and cytokeratin 18 (CK18) genes was measured by quantitative RT/PCR, and we have optimized this method by using SABiosciences qPCR Master Mix. We also employ an additional technique of immunomagnetic precipitation to extract RNA from CTCs. Specifically, we coat Dynabeads® (Invitrogen) with equal amounts of antibody to epithelial cell adhesion molecule (EpCAM) and to prostate specific membrane antigen (PSMA). RESULTS Using a specific primer/probe set, we demonstrate that our qRT/PCR assay is linear over a wide range, and that we can detect PSA transcript in RNA from < 1 LNCaP cell. When LNCaP cells are spiked into peripheral blood mononuclear cells (PBMNCs) from men without PCa, we demonstrate that we can detect 1 human prostate cell per 107 PBMNCs. In our phase II clinical trial we successfully detected CTCs in 13/17 (76.5%) patients. Our technique of dual antigen pull-down can detect 1 CTC per ml of whole blood of PCa patients. PSA transcripts have not been detected in negative control blood. CONCLUSIONS We have developed a novel, rapid and sensitive assay suitable for the detection of CTCs in peripheral blood of PCa patients. Table 1. Preliminary results in circulating tumor cell (CTC) detection in 17 patients, demonstrates successful detection in 13/17 (76.5%) patients. CTC Detection CTC Detection Patient PSA Stage Gleason PSA CK18 1 8.2 T1C 4+4 + NP 2 2.8 T2B/T3 5+4 + NP 3 4.1 T2B/T3 4+4 + NP 4 52 T1C 4+3 NEG NP 5 14 T2B/T3 3+4 NEG NP 6 4.9 T2A 4+4 + NP 7 5 T2B/T3 3+4 NEG NP 8 12 T1C 4+4 + NP 9 7.16 T1C 4+5 + NP 10 6.3 T2B 4+3 + NP 11 16.45 T2B 4+5 + NP 12 9.4 T1C 4+3 + NP 13 8.4 T2A 4+4 + NP 14 36 T3 4+3 NEG + 15 270 T4 4+4 NEG + 16 10 T2 4+4 NEG NEG 17 6.2 T1C 4+5 + + NP = Not performed, Neg = Negative. © 2011 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 185Issue 4SApril 2011Page: e54-e55 Advertisement Copyright & Permissions© 2011 by American Urological Association Education and Research, Inc.MetricsAuthor Information Xiaoke Huang Chicago, IL More articles by this author Barry B. McGuire Chicago, IL More articles by this author Irene Ogden Chicago, IL More articles by this author Daniel C. O'Brien Chicago, IL More articles by this author Phillip R. Cooper Chicago, IL More articles by this author Jessica A. Banks Chicago, IL More articles by this author William J. Catalona Chicago, IL More articles by this author Raymond C. Bergan Chicago, IL More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Transforming Growth Factor-Beta 1 Gene Polymorphisms and Expression in the Blood of Prostate Cancer Patients

The transforming growth factor beta 1 (TGF-β1) is a multifunctional cytokine with several regulatory activities in tumor cells affecting growth, differentiation, and function. Alterations in gene expression, secretion, and regulation of TGF-β1 may lead to a favorable environment for tumor development by angiogenesis stimulation and immune system suppression. We evaluated the influence of the TGFB1 polymorphisms by ARMS-PCR, Leu10Pro, and Arg25Pro, on prostate cancer (PCa) and benign prostatic hyperplasia (BPH). We assessed TGFB1 polymorphisms and their relation to mRNA levels (semi-quantitative RT-PCR) in blood samples as well as the implications in disease occurrence and progression. Peripheral blood samples from 175 patients were analyzed as to 92 BPH and 83 PCa. Samples obtained from 132 healthy males were used as negative controls. PCa patients with a Gleason score greater than 7 presented a higher frequency of the C allele (Leu10Pro). This allele was associated with a higher risk of developing PCa and BPH compared to the population (2.6 and 3.6 times higher, respectively). Patients with TGFB1 transcript levels equal to or more than 70% higher than control levels presented a 5.34 and 2.14-fold higher risk of having PCa and BPH, respectively, relative to the population. No association was detected between polymorphisms and mRNA levels. The C allele of the Leu10Pro polymorphism may predispose men to a more rapid cancer progression. Additionally, higher mRNA levels in the peripheral blood of PCa patients suggest that tumor cells may be disseminated in the circulation and could be used as a biomarker for extra-capsular invasion.