Gene of the month: NKX3.1

NKX3.1 is a multifaceted protein with roles in prostate development and protection from oxidative stress. Acting as a pioneer factor, NKX3.1 interacts with chromatin at enhancers to help integrate androgen regulated signalling. In prostate cancer, NKX3.1 activity is frequently reduced through a combination of mutational and post-translational events. Owing to its specificity for prostate tissue, NKX3.1 has found use as an immunohistochemical marker in routine histopathology practice.

Immunohistochemical Analysis of Prostein Expression in Archived Prostatic Core Biopsies from Prostate Cancer Patients in Western Kenya

Background Prostate cancer is the leading cause of cancer-associated mortality in men. Most of the current biomarkers for detection of the disease have low sensitivity and specificity. Prostein is a newly reported prostate cancer biomarkers whose diagnostic utility can help in early detection of the disease. Nonetheless, previous studies have utilized limited number of samples to evaluate its immunohistochemistry (IHC) and reports on the African population are not available. The current study aimed to determine the prostein expression in archived prostatic core biopsies from prostate cancer patients in Western Kenya. Materials and Methods This was a retrospective study conducted on malignant and benign prostatic tissue core biopsies of 106 patients who underwent prostate core biopsy at Jaramogi Oginga Odinga Teaching and Referral Hospital and division of urology at Synergy Clinics, Kisumu between January 2018 to May 2021. Immunohistochemical technique was performed on each of the 106 samples and on the following non-prostatic male control biopsies; Testis, Penis, Liver and Esophagus. Cellular location of prostein staining was evaluated at X40, X100 and X400 magnification using a light microscope and was classified as cytoplasmic or nucleocytoplasmic. Intensity of prostein expression was assessed for each core biopsy at similar magnification and graded according the immunohistochemistry composite score. Results The biopsies had been obtained from men whose mean (SE) age was 72.00±0.93 years. 95.3% (101) of the biopsies were malignant and 4.7% (5) were benign. Four non-prostatic male tissues were included. 97% of malignant and all the benign prostate tissue stained positive for prostein whereas the four non-prostatic male tissues were negative. Staining intensities were weak (24.5%), Moderate (17.0%), strong (55.7%) and non-stained (2.8%). The staining was highly immunolocalized within the cytoplasm (95.1% cases) as compared to nucleocytoplasmic (2.0% cases). The mean immunoreactivity composite score was 1.91±0.96 (0.0-3.14). Strongly stained sections had a punctate plasma membrane staining pattern clustered within the cytoplasm in a perinuclear location whereas the weakly stained sections had faint and punctate coarse brown cytoplasmic granular appearing. Conclusion Prostein is exclusively expressed in benign and malignant prostate tissue with a higher cytoplasmic granular staining pattern in the present population. These findings suggest that prostein diagnostic utility is applicable in the current study population and routine IHC diagnosis of prostate cancer may be recommended.

Dihydroartemisinin Promoted Bone Marrow Mesenchymal Stem Cell Homing and Suppressed Inflammation and Oxidative Stress against Prostate Injury in Chronic Bacterial Prostatitis Mice Model

Although bone marrow mesenchymal stem cells (BMMSCs) are effective in treating chronic bacterial prostatitis (CBP), the homing of BMMSCs seems to require ultrasound induction. Dihydroartemisinin (DHA) is an important derivative of artemisinin (ART) and has been previously reported to alleviate inflammation and autoimmune diseases. But the effect of DHA on chronic prostatitis (CP) is still unclear. This study aims to clarify the efficacy and mechanism of DHA in the treatment of CBP and its effect on the accumulation of BMMSCs. The experimental CBP was produced in C57BL/6 male mice via intraurethrally administered E. coli solution. Results showed that DHA treatment concentration-dependently promoted the accumulation of BMMSCs in prostate tissue of CBP mice. In addition, DHA and BMMSCs cotreatment significantly alleviated inflammation and improved prostate damage by decreasing the expression of proinflammatory factors such as TNF-α, IL-1β, and chemokines CXCL2, CXCL9, CXCL10, and CXCL11 in prostate tissue of CBP mice. Moreover, DHA and BMMSCs cotreatment displayed antioxidation property by increasing the production of glutathione peroxidase (GSH-Px), SOD, and decreasing malondialdehyde (MDA) expression. Mechanically, DHA and BMMSCs cotreatment significantly inhibited the expression of TGFβ-RI, TGFβ-RII, phosphor (p)-Smad2/3, and Smad4 in a dose-dependent manner while stimulated Smad7 expression in the same manner. In conclusion, our findings provided evidence that DHA effectively eliminated inflammatory and oxidative stress against prostate injury, and this effect involved the TGF-β/Smad signaling pathway in CBP.

Tissue stiffness in BPH patients from magnetic resonance elastography

Background BPH is commonly found in older men which can lead to lower urinary tract symptoms. Magnetic resonance elastography (MRE) is an innovative, noninvasive imaging technique used to evaluate tissue stiffness. There has not been any study, however, that assessed the tissue stiffness in patients with BPH. A prospective descriptive study was performed to demonstrated MRI and MRE techniques of the prostate gland in ten patients with BPH to assess tissue stiffness, features of BPH on MRI and components of BPH in the area of increased stiffness. Results MRI and MRE examinations in all patients were successful without any complications. The mean tissue stiffness of the whole prostate gland was 4.40 ± 0.71 kPa with good reproducibility (ICC 0.82). Stromal components and mixed glandular-stromal components tended to be associated with the areas of increased stiffness on stiffness images, 50.6% for stromal components and 37.9% for mixed glandular-stromal components. Some MRI findings were seen on the patients with high mean stiffness values such as prostatic calcification, type-5 BPH pattern and large prostate volumes. Conclusions Prostate MRE is a useful noninvasive reproducible diagnostic tool for evaluating prostate tissue stiffness by both qualitative and quantitative assessments. The mean prostate tissue stiffness from MRE in patients with BPH in this study was 4.40 ± 0.71 kPa. Some MRI features might be associated with increased tissue stiffness. Trial registration: PID 229. Registered 4 October 2019. http://md.redcap.kku.ac.th

Discovery proteomics defines androgen-regulated glycoprotein networks in prostate cancer cells, as well as putative biomarkers of prostatic diseases

Supraphysiologic androgen (SPA) inhibits cell proliferation in prostate cancer (PCa) cells by transcriptional repression of DNA replication and cell-cycle genes. In this study, quantitative glycoprotein profiling identified androgen-regulated glycoprotein networks associated with SPA-mediated inhibition of PCa cell proliferation, and androgen-regulated glycoproteins in clinical prostate tissues. SPA-regulated glycoprotein networks were enriched for translation factors and ribosomal proteins, proteins that are known to be O-GlcNAcylated in response to various cellular stresses. Thus, androgen-regulated glycoproteins are likely to be targeted for O-GlcNAcylation. Comparative analysis of glycosylated proteins in PCa cells and clinical prostate tissue identified androgen-regulated glycoproteins that are differentially expressed prostate tissues at various stages of cancer. Notably, the enzyme ectonucleoside triphosphate diphosphohydrolase 5 was found to be an androgen-regulated glycoprotein in PCa cells, with higher expression in cancerous versus non-cancerous prostate tissue. Our glycoproteomics study provides an experimental framework for characterizing androgen-regulated proteins and glycoprotein networks, toward better understanding how this subproteome leads to physiologic and supraphysiologic proliferation responses in PCa cells, and their potential use as druggable biomarkers of dysregulated AR-dependent signaling in PCa cells.

Chemical profiles of the active fraction from Prinsepia utilis Royle leaves and its anti-benign prostatic hyperplasia evaluation in animal models

Background The Prinsepia utilis Royle leaves (P. utilis) is a folk herb used for benign prostatic hyperplasia (BPH) control by ethnic minorities for centuries in China with rich in resources. Our previous studies have confirmed the anti-BPH effect of its water extract (QCJ) and the active fraction (Fr. B) separated from the QCJ by animal test. The Fr. B from P. utilis should be a potential candidate for BPH control. Methods In this study, the chemical ingredients of Fr. B were identified by UPLC-QTOF-MS, and quantified by HPLC. Murine animal models were divided into 8 groups, Sham rats, BPH rats, BPH rats administered with finasteride (1 mg/kg), BPH rats administered with Pule’an (460 mg/kg), BPH rats administered with low, high dosage of QCJ (860 mg/kg, 2580 mg/kg respectively), BPH rats administered with low, high dosage of Fr. B (160 mg/kg, 480 mg/kg respectively). The expression of vascular endothelial growth factor (VEGF) in the prostate tissue of rats was tested, and serum levels of dihydrotestosterone (DHT), testosterone (T), estradiol (E2), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and total superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT), malondialdehyde (MDA) in prostate homogenate were measured. One-way ANOVA followed by LSD was used for statistical analysis. Results The BPH rats treated by Fr. B exhibited significant reductions of VEGF and MDA levels, as well as significant increases of SOD, GSH-Px and CAT in the prostate tissue after 28 day administration (P < 0.05). Moreover, Fr. B significantly reduced DHT, DHT/E2 ratio, TNF-α, while increased T levels in serum of BPH rats (P < 0.05). UPLC-QTOF-MS analysis revealed 10 flavonoids as the key constituents of this fraction, which accounted for 54.96% of all substance of Fr. B. The relative contents of compound 1, 2 are 11.1%, 13% in Fr. B respectively. Conclusions These results indicated that the Fr. B obtained from P. utilis alleviated the symptoms of BPH rats through multiple mechanisms including reduction of DHT/E2 ratio, inhibition of growth factor, anti-inflammation and anti-oxidation, in which flavonoids might be the key constituents. It supported the hypothesis that the Fr. B should be further explored as a candidate for BPH patients.