Human Prostatic Cells Research Articles

IP-10 and CXCR3 signaling inhibit Zika virus replication in human prostate cells.

Our previous studies have shown that Zika virus (ZIKV) replicates in human prostate cells, suggesting that the prostate may serve as a long-term reservoir for virus transmission. Here, we demonstrated that the innate immune responses generated to three distinct ZIKV strains (all isolated from human serum) were significantly different and dependent on their passage history (in mosquito, monkey, or human cells). In addition, some of these phenotypic differences were reduced by a single additional cell culture passage, suggesting that viruses that have been passaged more than 3 times from the patient sample will no longer reflect natural phenotypes. Two of the ZIKV strains analyzed induced high levels of the IP-10 chemokine and IFNγ in human prostate epithelial and stromal mesenchymal stem cells. To further understand the importance of these innate responses on ZIKV replication, we measured the effects of IP-10 and its downstream receptor, CXCR3, on RNA and virus production in prostate cells. Treatment with IP-10, CXCR3 agonist, or CXCR3 antagonist significantly altered ZIKV viral gene expression, depending on their passage in cells of relevant hosts (mosquito or human). We detected differences in gene expression of two primary CXCR3 isoforms (CXCR3-A and CXCR3-B) on the two cell types, possibly explaining differences in viral output. Lastly, we examined the effects of IP-10, agonist, or antagonist on cell death and proliferation under physiologically relevant infection rates, and detected no significant differences. Although we did not measure protein expression directly, our results indicate that CXCR3 signaling may be a target for therapeutics, to ultimately stop sexual transmission of this virus.

In search of the correlation between nanomechanical and biomolecular properties of prostate cancer cells with different metastatic potential

Nanomechanical properties of living cells, as measured with atomic force microscopy (AFM), are increasingly recognized as criteria that differentiate normal and pathologically altered cells. Locally measured cell elastic properties, described by the parameter known as Young's modulus, are currently proposed as a new diagnostic parameter that can be used at the early stage of cancer detection. In this study, local mechanical properties of normal human prostate (RWPE-1) cells and a range of malignant (22Rv1) and metastatic prostate cells (LNCaP, Du145 and PC3) were investigated. It was found that non-malignant prostate cells are stiffer than cancer cells while the metastatic cells are much softer than malignant cells from the primary tumor site. Next, the biochemical properties of the cells were measured using confocal Raman (RS) and Fourier-transform infrared (FT-IR) spectroscopies to reveal these cells’ biochemical composition as malignant transformation proceeds. Nanomechanical and biochemical profiles of five different prostate cell lines were subsequently analyzed using partial least squares regression (PLSR) in order to identify which spectral features of the RS and FT-IR spectra correlate with the cell's elastic properties. The PLSR-based model could predict Young's modulus values based on both RS and FT-IR spectral information. These outcomes show not only that AFM, RS and FT-IR techniques can be used for discrimination between normal and cancer cells, but also that a linear correlation between mechanical response and biomolecular composition of the cells that undergo malignant transformation can be found. This knowledge broadens our understanding of how prostate cancer cells evolve thorough the multistep process of tumor pathogenesis.

Serenoa repens and Urtica dioica Fixed Combination: In-Vitro Validation of a Therapy for Benign Prostatic Hyperplasia (BPH).

Benign prostatic hyperplasia (BPH) is an age-related chronic disorder, characterized by the hyperproliferation of prostatic epithelial and stromal cells, which drives prostate enlargement. Since BPH aetiology and progression have been associated with the persistence of an inflammatory stimulus, induced both by Nuclear Factor-kappa B (NF-κB) activation and reactive oxygen species (ROS) production, the inhibition of these pathways could result in a good tool for its clinical treatment. This study aimed to evaluate the antioxidant and anti-inflammatory activity of a combined formulation of Serenoa repens and Urtica dioica (SR/UD) in an in vitro human model of BPH. The results confirmed both the antioxidant and the anti-inflammatory effects of SR/UD. In fact, SR/UD simultaneously reduced ROS production, NF-κB translocation inside the nucleus, and, consequently, interleukin 6 (IL-6) and interleukin 8 (IL-8) production. Furthermore, the effect of SR/UD was also tested in a human androgen-independent prostate cell model, PC3. SR/UD did not show any significant antioxidant and anti-inflammatory effect, but was able to reduce NF-κB translocation. Taken together, these results suggested a promising role of SR/UD in BPH and BPH-linked disorder prevention.

Abstract PO-022: The NKX3.1 homeoprotein functions in the mitochondria to protect against oxidative stress-dependent prostate tumorigenesis

Abstract The NKX3.1 homeobox gene is a key regulator of prostate differentiation, whose loss-of-function predisposes to prostate cancer. Here, we uncover an unexpected role for NKX3.1 in mitochondrial function and bioenergetics that regulates cancer initiation under conditions of oxidative stress. To investigate whether NKX3.1 plays a direct functional role in protection against oxidative stress, we studied the consequences of its loss- or gain-of-function in tumorigenic and non-tumorigenic human prostate cells, mutant mice, and primary human prostate cultures. Utilizing redox compounds to exogenously induce and study oxidative stress responses in our models, we show that NKX3.1 localizes to the mitochondria to protect against oxidative stress, while its loss-of-function leads to deregulation of mitochondrial-encoded oxidative phosphorylation genes, elevated reactive oxygen species, and accelerated prostate cancer initiation. Importantly, our studies reveal that cells lacking NKX3.1 have impaired intracellular energetics under oxidative stress, warranting further investigation into the role of NKX3.1 loss in cancer metabolism. These observations uncover a novel function for a tissue-specific differentiation regulator in mitochondria and demonstrate its functional relevance for suppressing prostate cancer initiation. Citation Format: Alexandros Papachristodoulou, Sukanya Panja, Antonina Mitrofanova, Aditya Dutta, Cory Abate-Shen. The NKX3.1 homeoprotein functions in the mitochondria to protect against oxidative stress-dependent prostate tumorigenesis [abstract]. In: Abstracts: AACR Special Virtual Conference on Epigenetics and Metabolism; October 15-16, 2020; 2020 Oct 15-16. Philadelphia (PA): AACR; Cancer Res 2020;80(23 Suppl):Abstract nr PO-022.

Inflammation driven tumor-like signaling in prostatic epithelial cells by sexually transmitted Trichomonas vaginalis.

To identify the sequence of inflammation-driven signaling cascades and other molecular events that might cause tumor-like transformation of prostatic cells. Cytokine array analysis, Reactome and STRING analysis, immunoblotting, and immunocytochemistry were used to investigate the molecular mechanisms governing inflammation-driven adverse changes in human prostatic cells caused by the sexually transmitted infection, Trichomonas vaginalis, resulting in prostatitis, benign prostatic hyperplasia and prostate cancer. Array analysis showed upregulation of 23 cytokines within 24h of infection of human prostatic epithelial RWPE-1 cells with the parasite, invitro. Reactome and STRING analysis of array data identified interleukin-6, interleukin-8, nuclear factor kappa B, signal transducer and activator of transcription3 and cyclooxygenase2 as chief instigators of prostatic anomaly, which were found to be significantly upregulated by immunofluorescence and western blotting analyses. STRING further connected these instigators with macrophage migration inhibitory factor, PIM-1 and prostate-specific antigen; which was confirmed by their marked stimulation in infected prostatic cells by immunoblotting and immunocytochemistry. Upregulated proliferation markers, such as Ki67, proliferating cell nuclear antigen and B-cell lymphoma2, suggested tumor-like signaling in infected RWPE-1 cells, which was further supported by downregulation of E-cadherin, upregulation of vimentin and activation of focal adhesion kinase. Prostate tumor DU145 cells were more sensitive to parasite invasion, and showed rapid upregulation with nuclear translocation of sensitive parameters, such as nuclear factor kappa B, signal transducer and activator of transcription3, and macrophage migration inhibitory factor. The migration of DU145 cells augmented when incubated in spent media from parasite-infected RWPE-1 cells. The initiation of inflammation driven tumor-like cell signaling in parasite-infected human prostatic epithelial cells is apparent, with the prostate tumor (DU145) cells being more sensitive to T. vaginalis than normal (RWPE-1) prostatic cells.

Investigating the performance of a novel pH and cathepsin B sensitive, stimulus-responsive nanoparticle for optimised sonodynamic therapy in prostate cancer

Nano-formulations that are responsive to tumour-related and externally-applied stimuli can offer improved, site-specific antitumor effects, and can improve the efficacy of conventional therapeutic agents. Here, we describe the performance of a novel stimulus-responsive nanoparticulate platform for the targeted treatment of prostate cancer using sonodynamic therapy (SDT). The nanoparticles were prepared by self-assembly of poly(L-glutamic acid-L-tyrosine) co-polymer with hematoporphyrin. The nanoparticulate formulation was characterized with respect to particle size, morphology, surface charge and singlet oxygen production during ultrasound exposure. The response of the formulation to the presence of cathepsin B, a proteolytic enzyme that is overexpressed and secreted in the tumour microenvironment of many solid tumours, was assessed. Our results showed that digestion with cathepsin B led to nanoparticle size reduction. In the absence of ultrasound, the formulation exhibited greater toxicity at acidic pH than at physiological pH, using the human prostate cells lines LNCaP and PC3 as targets. Nanoparticle cellular uptake was enhanced at acidic pH – a condition that was also associated with greater cathepsin B production. Nanoparticles exhibited enhanced ultrasound-induced cytotoxicity against both prostate cancer cell lines. Subsequent proof-of-concept in vivo studies demonstrated that, when ectopic human xenograft LNCaP tumours in SCID mice were treated with SDT using the systemically-administered nanoparticulate formulation at a single dose, tumour volumes decreased by up to 64% within 24 h. No adverse effects were observed in the nanoparticle-treated mice and their body weight remained stable. The potential of this novel formulation to deliver safe and effective treatment of prostate cancer is discussed.

Cytotoxic Effect of Saffron Stigma Aqueous Extract on Human Prostate Cancer and Mouse Fibroblast Cell Lines.

Several lines of experimental evidence have shown that saffron has anticarcinogenic effects. This study aimed at evaluating the possible anticancer effect of saffron stigma aqueous extract on human prostate cancer (PC3) and mouse fibroblast cells (L929) as non-cancerous control cells. Saffron stigma aqueous extract at concentrations of 100, 200, 400, 600, 800, 1600 and 3200 μg/mL were prepared. PC3 and L929 cells were incubated with different concentrations of saffron extracts in different time intervals (24, 48, 72, 96 and 144 hours). MTT assay was used for each cell line to investigate the cytotoxic effect of saffron. Morphological alterations were also observed under light inverted microscope. In fibroblast cell line after 24 hours, Saffron extract did not affect significantly the normal cells and they were intact in morphologic view. After 96 hours in the cells with highest concentration (1600 μg/mL), cell death and cellular form changes as well as severe granulation was observed. In prostate cell line after 24 hours, the only changes were observed in cells with the concentration of 1600 μg/mL. The cells were granulated and the form of the cells were spherule. After 72 hours, in group with the concentration of 1600 μg/mL, severe granulation was observed and the cell count decreased and some cells were dead. Saffron aqueous extract has an in vitro inhibitory effect on the proliferation of human prostate cell and mouse L929 cells which is dose-dependent.

New In Vitro Model of Oxidative Stress: Human Prostate Cells Injured with 2,2-diphenyl-1-picrylhydrazyl (DPPH) for the Screening of Antioxidants.

The antioxidant activity of natural compounds consists in their ability to modulate gene and protein expression, thus inducing an integrated cell protective response and repair processes against oxidative stress. New screening tools and methodologies are crucial for the actual requirement of new products with antioxidant activity to boost endogenous oxidative stress responsive pathways, Reactive Oxygen Species (ROS) metabolism and immune system activity, preserving human health and wellness. In this study, we performed and tested an integrated oxidative stress analysis, using DPPH assay and PNT2 cells injured with DPPH. We firstly investigated the mechanism of action of the oxidising agent (DPPH) on PNT2 cells, studying the variation in cell viability, oxidative stress genes, inflammatory mediator and ROS levels. The results reveal that DPPH activated ROS production and release of Prostaglandin E2 in PNT2 at low and intermediate doses, while cells switched from survival to cell death signals at high doses of the oxidising agent. This new in vitro oxidative stress model was validated by using Trolox, β-carotene and total extract of the green microalga Testraselmis suecica. Only the T. suecica extract can completely counteract DPPH-induced injury, since its chemical complexity demonstrated a multilevel protecting and neutralising effect against oxidative stress in PNT2.

Dextran-coated superparamagnetic nanoparticles modified with folate for targeted drug delivery of camptothecin

Drug delivery vehicles based on magnetic nanoparticles present a promising strategy for cancer treatment, due to controlled targeted treatment, high loading efficiency, and biosafety as compared to traditional drug delivery approaches. In the present study, superparamagnetic iron oxide (Fe3O4) nanoparticles (SPIONs) were synthesised by a co-precipitation approach, stabilised with dextran (DEX), and successfully conjugated with folate (FA) for delivery of camptothecin (CPT) in prostate cancer cells. Size and other characteristics of the modified nanoparticles were measured using scanning electron microscopy (SEM), transmission electron microscopy (TEM), dynamic light scattering (DLS), and zeta-potential. The results demonstrated that the prepared FA-DEX-SPIONs were spherical in shape with an average diameter of 63.31 nm. They had anticancer activity with high CPT loading efficiency in AT3B-1cancer cells, enabling the therapeutic activity of the drug via its active delivery and release at 37 °C in phosphate and citrate buffer solutions. MTT results exhibited no cytotoxicity on rat androgen independent prostate cancer (AT3B-1) and normal human prostate (RWPE-1) cells. In conclusion, FA-DEX-SPION composite is a promising candidate that could be used for the targeted release of CPT anti-cancer drug.

Targeting Androgen Regulation of TMPRSS2 and ACE2 as a Therapeutic Strategy to Combat COVID-19

The COVID-19 pandemic is expected to have an adverse effect on the progression of multiple cancers, including prostate cancer, due to the ensuing cytokine storm and associated oncogenic signaling. Epidemiological data showing increased severity and mortality of COVID-19 in men suggests a potential role for androgen in SARS-CoV-2 infection. Here, we present evidence for the transcriptional regulation of SARS-CoV-2 host cell receptor ACE2 and co-receptor TMPRSS2 by androgen in mouse tissues and human prostate and lung cell lines. Additionally, we demonstrate the endogenous interaction between TMPRSS2 and ACE2 in human cells and validate ACE2 as a TMPRSS2 substrate. In an overexpression model, and the prostate and lung cells, Camostat – a TMPRSS2 inhibitor, blocked the cleavage of pseudotype SARS-CoV-2 surface Spike without disrupting TMPRSS2-ACE2 interaction. Thus providing evidence for the first time a direct role of TMPRSS2 in priming the SARS-CoV-2 Spike protein, required for viral fusion to the host cell. Importantly, androgen-deprivation, anti-androgens such as enzalutamide/AR-PROTAC, or Camostat treatment attenuated the SARS-CoV-2 S-mediated entry in lung and prostate cells. Together, our preclinical data provide a strong rationale for clinical evaluations of the TMPRSS2 inhibitors, androgen-deprivation therapy and androgen receptor antagonists alone or in combination with anti-viral drugs as early as clinically possible to prevent inflammation driven COVID-19 progression.Funding: Research in I.A.A laboratory is supported by NIH (1-R01 CA249210-0), Department of Defense Idea Development Award (W81XWH-17-0404), and Sarcoma Foundation of America Research Award to I.A.A.Ethical Approval: In vivo mouse studies were performed following protocols approved by the Institutional Animal Care and Use Committee at the University of Pennsylvania and in compliance with all regulatory standards. C57BL/6J mice were procured from Jackson Laboratory (000664) and breed in house.Declaration of Interest: None to declare

MiRNA-191 promotes proliferation, migration and invasion of prostate cancer by targeting PLCD1

Objective: To investigate the effects of miRNA-191 on the proliferation, migration and invasion of prostate cancer, and to explore its mechanism. Methods: The expression levels of miRNA-191 in four human prostate cancer cell lines (PC-3, DU-145, LNCa P, 22RU1) and human normal prostate cell line RWPE-2 were detected, and prostate cancer cell line PC-3 was selected as the experimental object. PC-3 cells were divided into three groups: blank control group (no transfection), miRNA-191 NC group (PC-3 cells transfected with Inhibitor NC) and miRNA-191 Inhibitor group (PC-3 cells transfected with miRNA-191 Inhibitor), and each group was provided with three multiple pores. The expression levels of miRNA-191 and PLCD1 were detected by RT-PCR. The cell proliferation was detected by CCK8 assay. Scratch test and invasive test were used to detect cell migration and invasive ability. Through Targetscan target gene prediction website, PLCD1 was screened as the target protein of miRNA-191, and verified by double luciferase target experiment.Western blot assay was used to detect the expression of PLCD1 in cells of each group. Results: Compared with RWPE-2 cells, the expression level of miRNA-191 in human prostate cancer cells was significantly higher (P ＜0.05), and the expression level of miRNA191 in PC-3 was significantly higher than that in other three cell lines (P＜0.05). After inhibiting the expression of miRNA-191, the expression levels of PLCD1 was significantly higher while PC-3 cells' proliferation ability was inhibited, and their migration and invasion ability were significantly lower than those of blank control group and miRNA-191 NC group (P＜ 0.05). The results of double luciferase reporter gene assay showed that PLCD1 gene was a target gene of miRNA-191. Conclusion: miRNA-191 promote the proliferation, migration and invasion of prostate cancer PC-3 cells by targeting PLCD1.

Estimation of mandarin peel oil–induced cytotoxicity and genotoxicity in human normal fibroblast and cancerous prostate cell lines

The low price and high contents of bioactive compounds in citrus peel increase interest in using it in various applications. Mandarin (Citrus reticluata) peel belongs to Rutaceae family and is rich with antioxidants. However, limited studies are available on toxicity of Mandarin peel oil (MPO) on human Prostate Cancer (PC3) cells. Therefore, the present study was conducted to study the cytotoxicity and genotoxicity of MPO on Human normal Fibroblast (HFB4) and PC3 cell lines. The half maximal inhibitory (IC50) and safe concentration of MPO was detected using MTT assay. Comet and DNA fragmentation assays were performed to assess apoptotic DNA damage. Also, the ROS level was evaluated and the mRNA expression level of apoptotic and antiapoptotic genes were measured using RT-PCR. Results of the cytotoxic test showed that MPO induced preferential inhibition of PC3 cells proliferation in a concentration-dependent manner with IC50 10.97 µg/ml. The time-dependent induction of DNA breaks demonstrated in PC3 cells treated with MPO safe concentration-stimulated ROS generation and apoptotic DNA damage through increased expression of tumor suppressor p53 and Bax genes and decreased expression of Bcl2 and MDM2 genes. In contrast, non-significant changes were observed in the DNA integrity, ROS levels and expressions of the tested genes in the normal HFB4 cells treated with MPO. Thus, we concluded that MPO induced preferential cytotoxic and genotoxic effects toward cancerous PC3 with no noticeable toxic effects in normal HFB4 cells and therefore further in vivo studies are recommended to test its possible use as anticancer drugs.

Peritumoral Delivery of Docetaxel-TIPS Microparticles for Prostate Cancer Adjuvant Therapy.

Recurrence of prostate cancer after radical prostatectomy is a consequence of incomplete tumor resection. Systemic chemotherapy after surgery is associated with significant toxicity. Improved delivery methods for toxic drugs capable of targeting positive resection margins can reduce tumor recurrence and avoid their known toxicity. This study evaluates the effectiveness and toxicity of docetaxel (DTX) release from highly porous biodegradable microparticles intended for delivery into the tissue cavity created during radical prostatectomy to target residual tumor cells. The microparticles, composed of poly(dl‐lactide‐co‐glycolide) (PLGA), are processed using thermally induced phase separation (TIPS) and loaded with DTX via antisolvent precipitation. Sustained drug release and effective toxicity in vitro are observed against PC3 human prostate cells. Peritumoral injection in a PC3 xenograft tumor model results in tumor growth inhibition equivalent to that achieved with intravenous delivery of DTX. Unlike intravenous delivery of DTX, implantation of DTX‐TIPS microparticles is not accompanied by toxicity or elevated systemic levels of DTX in organ tissues or plasma. DTX‐TIPS microparticles provide localized and sustained release of nontoxic therapeutic amounts of DTX. This may offer novel therapeutic strategies for improving management of patients with clinically localized high‐risk disease requiring radical prostatectomy and other solid cancers at high risk of positive resection margins.

CHD1 and SPOP synergistically protect prostate epithelial cells from DNA damage.

Recent genomic profiling has identified a subtype of prostate cancer (PCa) characterized by two key genetic alterations: missense mutation of speckle-type POZ protein (SPOP) and homozygous deletion of chromodomain helicase DNA-binding protein 1 (CHD1). Mutually exclusive with E26 transformation-specific (ETS) rearrangements, this subtype displays high genomic instability. Previous studies indicate that deficient SPOP or CHD1 alone leads to feeble prostate abnormalities and each protein is involved in DNA damage response (DDR). It remains to be determined whether CHD1 and SPOP cooperate to suppress prostate tumorigenesis and DDR. Prostate-specific single or double knockout of Spop and Chd1 was generated with the Cre/loxP system in mice. Wild-type or mutant SPOP (F102C, F133V) overexpression and CHD1 knockdown with short hairpin RNA were created in human benign prostatic hyperplasia cell line BPH1. The levels of DNA damage and homologous recombination repair were measured by immunofluorescence staining of γH2AX and RAD51, respectively. Spop/Chd1 double-knockout mice displayed prostatic intraepithelial neoplasia at both young (3 months) and old (12 months) ages and failed to generate prostate adenocarcinoma. Compared with wild-type or single-knockout mice, the double-knockout prostate harbored moderately higher proliferating cells and dramatically augmented the level of γH2AX staining, although androgen receptor-positive cells and apoptotic cells remained at a similar level. In BPH1 cell line, SPOP mutant overexpression and CHD1 silencing synergistically sensitized the cells to DNA damage by camptothecin, an inducer of double-strand breaks. Our results indicate that SPOP and CHD1 can synergistically promote repair of naturally occurring or chemically induced DNA damages in prostate epithelial cells. Regarding the progression of the SPOP/CHD1 subtype of PCa, other functionally complementary drivers warrant further identification. The clinical implication is that this subtype of PCa may be particularly sensitive to poly(ADP-ribose) polymerase inhibitors or DNA-damaging agents.

Effects of Quercetin on Autophagy and Phosphatidylinositol 3-kinase/Protein Kinase B/Mammalian Target of Rapamycin Signaling Pathway in Human Prostate Cancer PC-3 Cells

Objective To investigate the effects of quercetin on cell viability,apoptosis,autophagy,and phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR)signaling pathway in human prostate cell carcinoma PC-3 cells.Methods PC-3 cells were cultured in vitro,and cell viability was detected by CCK-8.Apoptosis was detected by TUNEL staining.Autophagy vesicle was observed by acridine orange staining.Autophagosomes was observed by GFP-LC3 plasmid transfection analysis.Expressions of autophagy-related protein microtubule associated protein 1 light chain 3 fusion protein(LC3)and Beclin-1 and PI3K/Akt/mTOR signaling pathway protein were detected by Western blot analysis.Results Quercetin inhibited cell viability in a dose-time dependent manner and induced apoptosis.Quercetin increased the number of autophagy vesicles and autophagosomes in PC-3 cells.Quercetin increased the expressions of LC3-Ⅱ/LC3-Ⅰ and Beclin-1 in PC-3 cells and decreased the expression of phosphorylated-PI3K,phosphorylated-Akt and phosphorylated-mTOR.Conclusion Quercetin may induce autophagy by inactivating PI3K/Akt/mTOR signaling pathway in PC-3 cells.

Establishment and characterization of a docetaxel‑resistant human prostate cancer cell line

The aim of the present study was to establish a novel docetaxel-resistant prostate cancer cell line and investigate its biological characteristics. The human prostate cell line, PC-3, was exposed to docetaxel, the concentrations of which were increased in a stepwise manner in the medium to select the drug-resistant cell line, PC-3/DTX. The morphological features were observed using inverted microscopy. The growth curves of PC-3 and PC-3/DTX cells were drawn to calculate the doubling time. Flow cytometry was performed to determine cell-cycle distribution. A 3-(4,5-dimethyl-2-thiazol)-2,5-diphenyl-2H tetrazolium bromide assay was performed to test the drug resistance of PC-3 and PC-3/DTX cells. Western blot analysis was conducted to determine the protein expression levels of the mammalian target of rapamycin (mTOR) signaling pathway, which may serve a role in regulating drug resistance in the two cell lines. PC-3/DTX cells exhibited changes in morphology, proliferation rate, doubling time and cell-cycle distributions, compared with PC-3 cells. PC-3/DTX cells were 10.9-fold resistant to docetaxel in comparison with PC-3 cells. The results showed that PC-3/DTX cells overexpressed Rictor and p-AKT(S473) proteins, which are specific subunits or downstream substrates of mTORC2. The new findings suggested that the mTORC2 signaling pathway may serve an important role in the regulation of docetaxel drug resistance of PC-3 cells. In conclusion, PC-3/DTX cells may be applied to study the resistance of anticancer drugs and to identify methods to overcome resistance.

TRIM52 positively mediates NF-κB to promote the growth of human benign prostatic hyperplasia cells through affecting TRAF2 ubiquitination

AimsBenign prostatic hyperplasia (BPH) is a progressive disease, which severely affects men's health. Here, we sought to analyze the functions and mechanism of action of the tripartite motif protein 52 (TRIM52), a novel prostate basal cell biomarker in BPH. Materials and methodsImmunohistochemistry assay was performed in sectioned human BPH tissues, BPH-1 cells, and prostate RWPE-1 cells, to detect the expressions of TRIM52 and NF-κB. Western blotting and qRT-PCR analyses were conducted to measure the relative protein and mRNA expression levels, respectively. Further, lentiviral transfection was performed in BPH-1 and RWPE-1 cells to study the overexpression and siRNA knockdown of TRIM52. Dual-luciferase reporter assay was applied to evaluate the relationship between NF-κB and TRIM52. Furthermore, CCK-8 assay and flow cytometry were employed to analyze cell proliferation and apoptosis. Key findingsTRIM52 and NF-κB levels were elevated in BPH tissues, and TRIM52 expression positively correlated with NF-κB expression. TRIM52 silencing suppressed the growth of BPH-1 cells and decreased the promoter activity of NF-κB. Moreover, the NF-κB inhibitor, pyrrolidine dithiocarbamate (PDTC), suppressed TRIM52-induced proliferation of RWPE-1 cells and inhibited NF-κB promoter activity in oeTRIM52 transfected RWPE-1 cells. Silencing TRIM52 also inhibited TRAF2 ubiquitination in BPH-1 cells. Further, NF-κB promoter activity in siNC transfected cells was enhanced by the recombinant protein TNF-α and inhibited by siTRIM52. SignificanceTRIM52 accelerated the growth of BPH-1 cells by upregulating NF-κB, and TRIM52 could promote TRAF2 ubiquitination. These findings might contribute to the understanding of the biological functions and action mechanisms of TRIM52 in BPH.

Does Bromelain-Cisplatin Combination Afford In-Vitro Synergistic Anticancer Effects on Human Prostatic Carcinoma Cell Line, PC3?

Background: Bromelain enhances anticancer impacts to chemotherapeutic agents. The question as to whether bromelain does promote in-vitro cytotoxic and proapoptotic effects of cisplatin on human prostatic carcinoma PC3 cell line was investigated. Materials and Methods: PC3 (human prostatic carcinoma) cells were treated either single or in combination with bromelain and/or cisplatin. MTT, clonogenic assay, flow cytometry and real-time quantitative polymerase chain reaction were used to investigate cell viability, colony formation, proapoptotic potential and p53 gene expression, respectively. Results: Cisplatin (IC10) combined with bromelain (IC40) significantly affected PC3 cell viability, inhibited colony formation, as well increased p53 proapoptotic gene expression compared to cisplatin single treatment. Nevertheless, bromelain-cisplatin chemoherbal combination did not display any additive proapoptotic effect compared to single treatments. Conclusion: Bromelain-cisplatin chemoherbal combination demonstrated synergistic in-vitro anticancer effect on human prostatic carcinoma cell line, PC3, that drastically reduced required cisplatin dose.

Does Bromelain-Cisplatin Combination Afford In-Vitro Synergistic Anticancer Effects on Human Prostatic Carcinoma Cell Line, PC3?

Background: Bromelain enhances anticancer impacts to chemotherapeutic agents. The question as to whether bromelain does promote in-vitro cytotoxic and proapoptotic effects of cisplatin on human prostatic carcinoma PC3 cell line was investigated. Materials and Methods: PC3 (human prostatic carcinoma) cells were treated either single or in combination with bromelain and/or cisplatin. MTT, clonogenic assay, flow cytometry and real-time quantitative polymerase chain reaction were used to investigate cell viability, colony formation, proapoptotic potential and p53 gene expression, respectively. Results: Cisplatin (IC10) combined with bromelain (IC40) significantly affected PC3 cell viability, inhibited colony formation, as well increased p53 proapoptotic gene expression compared to cisplatin single treatment. Nevertheless, bromelain-cisplatin chemoherbal combination did not display any additive proapoptotic effect compared to single treatments. Conclusion: Bromelain-cisplatin chemoherbal combination demonstrated synergistic in-vitro anticancer effect on human prostatic carcinoma cell line, PC3, that drastically reduced required cisplatin dose. [GMJ.2020;9:e1749]

Single-cell transcriptomics identifies a distinct luminal progenitor cell type in distal prostate invagination tips.

The identification of prostate stem/progenitor cells and characterization of the prostate epithelial cell lineage hierarchy are critical for understanding prostate cancer initiation. Here, we characterized 35,129 cells from mouse prostates, and identified a unique luminal cell type (termed type C luminal cell (Luminal-C)) marked by Tacstd2, Ck4 and Psca expression. Luminal-C cells located at the distal prostate invagination tips (termed Dist-Luminal-C) exhibited greater capacity for organoid formation in vitro and prostate epithelial duct regeneration in vivo. Lineage tracing of Luminal-C cells indicated that Dist-Luminal-C cells reconstituted distal prostate luminal lineages through self-renewal and differentiation. Deletion of Pten in Dist-Luminal-C cells resulted in prostatic intraepithelial neoplasia. We further characterized 11,374 human prostate cells and confirmed the existence of h-Luminal-C cells. Our study provides insights into the prostate lineage hierarchy, identifies Dist-Luminal-C cells as the luminal progenitor cell population in invagination tips and suggests one of the potential cellular origins of prostate cancer.