Human Prostate Carcinoma Cells Research Articles

Effect of Roasting Levels and Drying Process of Coffea canephora on the Quality of Bioactive Compounds and Cytotoxicity.

Coffee is a popular drink consumed all over the world. Besides its long-recognized stimulant effect, it has important nutritional and health effects. However, the type of bean processing modifies the composition of brewed coffee and possibly its bioactivity. In this study, extracts obtained from green and roasted beans of Coffea canephora (Coffea canephora var. robusta) were submitted to spray- or freeze-drying and were tested for antiproliferative activity, using MTT assay, and their influence on the cell cycle and apoptosis by flow cytometry analysis. Moreover, colors and nutrient contents were measured to identify the changes due to the roasting process. The results obtained showed that extracts from green and light roasted beans exhibited strong bioactive capacity. Coffee extracts promoted a decrease in cell viability, modulated cell cycle and induced apoptosis in human prostate carcinoma cell line (DU-145). The level of roasting reduced this property, but the type of drying did not in all cases.

Anticancer activity evaluation of green synthesised gold–silver alloy nanoparticles on colourectal HT‐29 and prostate DU‐145 carcinoma cell lines

Nanotechnology is an interesting way to produce and deal with fascinating features of materials. Metal nanoparticles offer a vast spectrum of potential applications in medicine and pharmaceutical sciences due to advances in nanotechnology research. This study was aimed to evaluate the anticancer properties of gold–silver (Au–Ag) alloy nanoparticles, which were synthesised using a green method by Galega officinalis ethanolic extract. The antitumour activity of biosynthesised nanoparticles has been assessed in vitro using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay on DU-145 and HT-29, prostate and colon cell lines, respectively. The Au–Ag alloy nanoparticles activity to inhibit the growth of cancer cell lines was recorded in terms of decrease in viable cell count in comparison with the control value. The spherical Au–Ag alloy nanoparticles with average size of 100 nm diameter were synthesised. The growth inhibition of the human prostate (DU-145) and colon carcinoma cell line (HT-29) has been found to be dose dependent. The calculated IC50 for the DU-145 cell line was 95.78 μg/ml after 24 h and for the HT-29 cell line was 70.66 µg/ml after 48 h. The results of this study suggest that Au–Ag alloy nanoparticles could be as an active substance against tumour cell lines and may be a beneficial treatment for cancer.

IRS-2 deubiquitination by USP9X maintains anchorage-independent cell growth via Erk1/2 activation in prostate carcinoma cell line.

Insulin-like growth factors (IGFs) have been shown to induce proliferation of many types of cells. Insulin receptor substrates (IRSs) are major targets of IGF-I receptor (IGF-IR) tyrosine kinase activated by IGFs, and are known to play important roles in the activation of downstream signaling pathways, such as the Erk1/2 pathway. Dysregulation of IGF signaling represents a central tumor promoting principle in human carcinogenesis. Prostate carcinoma is highly dependent on the IGF/IGF-IR/IRS axis. Here we identified the deubiquitinase, ubiquitin specific peptidase 9X (USP9X) as a novel binding partner of IRS-2. In a human prostate carcinoma cell line, small interfering RNA (siRNA)-mediated knockdown of USP9X reduced IGF-IR as well as IRS-2 protein levels and increased their ubiquitination. Knockdown of USP9X suppressed basal activation of the Erk1/2 pathway, which was significantly restored by exogenous expression of IRS-2 but not by IGF-IR, suggesting that the stabilization of IRS-2 by USP9X is critical for basal Erk1/2 activation. Finally, we measured anchorage-independent cell growth, a characteristic cancer feature, by soft-agar colony formation assay. Knockdown of USP9X significantly reduced anchorage-independent cell growth of prostate carcinoma cell line. Taken all together, our findings indicate that USP9X is required for the promotion of prostate cancer growth by maintaining the activation of the Erk1/2 pathway through IRS-2 stabilization.

Cytotoxicity of AMANTADIG – a semisynthetic digitoxigenin derivative – alone and in combination with docetaxel in human hormone-refractory prostate cancer cells and its effect on Na+/K+-ATPase inhibition

Cardiac glycosides (CGs) are natural compounds used to treat congestive heart failure. They have garnered attention as a potential cancer treatment option, especially because they bind to Na+/K+-ATPase as a target and activate intracellular signaling pathways leading to a variety of cellular responses. In this study we evaluated AMANTADIG, a semisynthetic cardenolide derivative, for its cytotoxic activity in two human androgen-insensitive prostate carcinoma cell lines, and the potential synergistic effects with docetaxel. AMANTADIG induced cytotoxic effects in both cell lines, and a combination with docetaxel showed a moderate and strong synergism in DU145 and PC-3 cells, respectively, at concentrations considerably lower than their IC50 values. Cell cycle analyses showed that AMANTADIG and its synergistic combination induced G2/M arrest of DU145 and PC-3 cells by modulating Cyclin B1, CDK1, p21 and, mainly, survivin expression, a promising target in cancer therapy. Furthermore, AMANTADIG presented reduced toxicity toward non-cancerous cell type (PBMC), and computational docking studies disclosed high-affinity binding to the Na+/K+-ATPase α subunit, a result that was experimentally confirmed by Na+/K+-ATPase inhibition assays. Hence, AMANTADIG inhibited Na+/K+-ATPase activity in PC-3 cells, as well as in purified pig kidney at nanomolar range. Altogether, these data highlight the potent effects of AMANTADIG in combination with docetaxel and offer important insights for the development of more effective and selective therapies against prostate cancer.

Quantitative microinjection using fluorescence calibration of streaming microdroplets on a superhydrophobic surface

A simple and reproducible procedure was developed to measure the volume of liquid microinjected into cells. A calibration curve of droplet fluorescence intensity versus volume was constructed by injecting a fluorescent dextran solution through a 125–150 µm diameter micropipette into an oil-filled culture dish to create a spray of varied-sized droplets. The droplets retained a spherical shape because they were in an oil medium and they settled onto a glass surface coated with a superhydrophobic surface. Fluorescent micrographs of the droplets were obtained and analyzed with Image-J software to quantify the fluorescence intensity and radius of each spherical droplet to produce the calibration curve. Subsequently, Dut-145 human prostate carcinoma cells were microinjected with the same fluorescent dextran solution and fluorescent micrographs of the cells were obtained using the identical exposure conditions used to photograph the droplets. The measured fluorescence intensity of the microinjected cells was entered into the formula for the regression line that was fit to the calibration curve allowing determination of the volume of solution injected into each cell. Thus, a mixture consisting of known concentrations of a test material of test material (macromolecules, drugs, etc.) and a fluorescent dextran, volumetric, tracer can be used to quantify the relationship between the amount of a microinjected material and subsequent effects on cells.

Nintedanib inhibits growth of human prostate carcinoma cells by modulating both cell cycle and angiogenesis regulators

Prostate cancer (PCa) is the most common malignancy and second leading cause of cancer-related deaths in American men. Proliferating cells have higher need for nutrients and oxygen, triggering angiogenesis that plays a critical role in tumor growth, progression and metastasis. Consequently, immense focus has converged onto inhibitors of angiogenesis in cancer treatment, such as Nintedanib, which has shown exceptional antitumor activity via inhibiting cell proliferation and the resulting tumor growth, primarily due to its combined action on tumor cells, endothelial cells and pericytes. Accordingly, here we assessed both in vitro and in vivo efficacy of Nintedanib in PCa. The results showed that Nintedanib decreased cell viability in both androgen dependent- and -independent PCa cells, together with a decrease in cell motility and invasiveness. Nintedanib also reduced the expression of significant genes responsible for cell cycle progression. PCa PC3 xenograft-carrying nude mice treated with Nintedanib showed significantly decreased tumor volume and cell proliferation alongside diminished levels of pro-angiogenic molecules and blood vessel densities. In conclusion, we report that Nintedanib has strong efficacy against PCa in pre-clinical models via modulation of various pathways, and that it could be employed as a promising new strategy to manage PCa clinically.

Inhibition of ANO1/TMEM16A induces apoptosis in human prostate carcinoma cells by activating TNF-\u03b1 signaling

Overexpression of the Ca2+-activated chloride channel ANO1/TMEM16A is implicated in tumorigenesis, and inhibition of ANO1 overexpression suppresses xenograft tumor growth and invasiveness. However, the underlying molecular mechanism for ANO1 inhibition in suppression of tumorigenesis remains unknown. Here, we show that silencing or inhibition of endogenous ANO1 inhibits cell growth, induces apoptosis and upregulates TNF-α expression in prostate cancer PC-3 cells. Enhancement of TNF-α signaling by ANO1 knockdown leads to upregulation of phosphorylated Fas-associated protein with death domain and caspase activation. Furthermore, silencing of ANO1 inhibits growth of PC-3 xenograft tumors in nude mice and induces apoptosis in tumors via upregulation of TNF-α signaling. Taken together, our findings provide mechanistic insight into promoting apoptosis in prostate cancer cells by ANO1 inhibition through upregulation of TNF-α signaling.

Procyanidin from peanut skin induces antiproliferative effect in human prostate carcinoma cells DU145

In this study, the antiproliferative activity of peanut skin procyanidins (PSP) and six fractions (PSP-1∼6) isolated from PSP by several chromatographic steps on the human prostate cancer DU145 cells were evaluated. The results showed that PSP and PSP-1∼6 significantly inhibited the proliferation of DU145 cells. PSP-2 was the most effective fraction, which was identified as procyanidin B3 mainly and procyanidin dimer [(E)C-luteolin or keampferol] secondarily. Moreover, the mechanism of antiproliferative activity of PSP-2 was investigated. It was observed that PSP-2 induced apoptotic cell death and cell cycle arrest at S phase in DU145 cells. PSP-2 caused the increase of intracellular ROS level and the decrease of Bcl-2/Bax ratio, and triggered the activation of p53 and caspases-3 in DU145 cells. Our findings demonstrated that procyanidins from peanut skin have the potential to be developed as an anti-prostate cancer agent.

Chemotherapeutic Effect of CD147 Antibody-labeled Micelles Encapsulating Doxorubicin Conjugate Targeting CD147-Expressing Carcinoma Cells.

CD147 (basigin/emmprin) is expressed on the surface of carcinoma cells. For studying the efficacy of CD147-targeting medicine on CD147-expressing cells, we studied the effect of anti-CD147-labeled polymeric micelles (CD147ab micelles) that encapsulated a conjugate of doxorubicin with glutathione (GSH-DXR), with specific accumulation and cytotoxicity against CD147-expressing A431 human epidermoid carcinoma cells, Ishikawa human endometrial adenocarcinoma cells, and PC3 human prostate carcinoma cells. By treatment of each cell type with CD147ab micelles for 1 h, a specific accumulation of CD147ab micelles in CD147-expressing cells was observed. In addition, the cytotoxicity of GSH-DXR-encapsulated micelles against each cell type was measured by treatment of the micelles for 1 h. The cytotoxic effect of CD147ab micelles carrying GSH-DXR was 3- to 10-fold higher for these cells than that of micelles without GSH-DXR. These results suggest that GSH-DXR-encapsulated CD147ab micelles could serve as an effective drug delivery system to CD147-expressing carcinoma cells.

Major terpenoids from Telekia speciosa flowers and their cytotoxic activity in vitro

In addition to known constituents of Telekia speciosa, an acetone extract from ray florets of the plant yielded: 5,5ʹ-dibutoxy-2,2ʹ-bifuran (1), 5,5ʹ-diisobutoxy-2,2ʹ-bifuran (2), α-tocopherol (3), β-tocopherol (4), loliolide palmitate (5), a mixture of calenduladiol esters - 16β-hydroxylupeol-3-O-palmitate (7) and 16β-hydroxylupeol-3-O-myristate (8), 1-epiinuviscolide (12), inuviscolide (13), 3-epiisotelekin (16), 4α-hydroxy-9β,10β-epoxy-1β(H)-11(13)-guaien-8α,12-olide (17), 4α-hydroxy-1β(H)-9(10),11(13)-guaiadien-8α,12-olide (18), loliolide (19) and 4β,10β-dihydroxy-1α(H),5α(H)-11(13)-guaien-8α,12-olide (20). Calenduladiol esters and asperilin (14) were the major constituents of the extract. Their cytotoxic effect on human normal prostate epithelial cells (PNT-2), human prostate carcinoma cell lines, human skin fibroblasts (HSF) and human melanoma cell lines was examined in vitro. Triterpene esters showed no cytotoxicity against nearly all cell lines tested, except for Du145 prostate carcinoma cells (IC50 – 62.0 μΜ). Asperilin displayed activity against the cell lines under study, especially against three tested lines of melanomas (A375, IC50 – 17.6 μΜ, WM793, IC50 – 28.2 μΜ and Hs 294T, IC50 – 29.5 μΜ).

Potential anticancer activity of biogenic silver nanoparticles using leaf extract of Rhynchosia suaveolens: an insight into the mechanism

The present study reports a simple and eco-friendly synthesis of silver nanoparticles (AgNPs) using leaf extract of Rhynchosia suaveolens. UV-Vis analysis of R. suaveolens synthesized AgNPs (RS-AgNPs) showed surface plasmon resonance (SPR) peak at 426 nm. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) analysis revealed that RS-AgNPs were 10–30 nm in size with spherical shape. X-ray diffraction (XRD) analysis of RS-AgNPs confirmed the crystalline nature with face-centered cubic (FCC) lattice. Fourier transform infrared (FTIR) interprets that polyphenols and proteins take part in bioreduction and capping of RS-AgNPs. RS-AgNPs exhibited dose-dependent inhibition of proliferation of different cancer cells including DU145 and PC-3(human prostate carcinoma cell lines), SKOV3 (human ovarian carcinoma) and A549 (human lung adenocarcinoma)with IC50 values of 4.35, 7.72, 4.2 and 24.7 μg/mL, respectively. The plausible reasons behind anticancer activity of RS-AgNPs were explained using different assays on the most susceptible SKOV3 cells. RS-AgNPs induced oxidative stress in SKOV3 cells by generating reactive oxygen species (ROS), enhancing lipid peroxidation (LPO) levels and decreasing glutathione (GSH) levels. RS-AgNPs induced the apoptosis of SKOV3 cells by up regulating the caspase-3, caspase -8, caspase -9, p53 and BAX and down regulating the antiapoptotic protein Bcl-2. Further, RS-AgNPs showed elevation of caspase 3/7 activity and also exhibited antimigratory effect by inhibiting the migration of SKOV3 cells into the wounded area. The findings suggested that biogenic RS-AgNPs provide an alternative approach to overcome several limitations of chemotherapy.

Flavonolignans and Biological Activity of Nannorrhops ritchiana Leaves

Objectives: This study aimed at isolating the polyphenolic compounds of 70% methanol extract ofNannorrhops ritchiana Griff. leaves and assaying the antioxidant and cytotoxic activities of the extract and main fractions. Methods: The methanol extract of the leaves of N. ritchiana was chromatographically fractionated using a bioactivity-guided approach. The isolated compounds were spectroscopically elucidated by UV, MS, 1H/13C NMR and 2D NMR spectroscopic techniques. The radical scavenging activity of the methanol extract and its fractions was evaluated using DPPH (2,2-Diphenyl-1-picrylhydrazyl) radical scavenging assay and their cytotoxic activity was assayed using SRB (3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyltetrazolium bromide) test against human hepatocellular liver carcinoma (HepG2), human alveolar adenocarcinoma (A549) and human prostate carcinoma (PC3) cell lines. Results: Eight compounds were isolated and identified for the first time from the leaves of N. ritchiana, comprising five flavonolignans;Indocalatin A (4) which is a rare compound that has been reported only once in nature, 9″-O-glucopyranosyl Salcolin A (5), 9″-O-glucopyranosyl Salcolin B (6) together with their aglycones, Salcolin A (7) and Salcolin B (8) along with three flavone glycosides; Tricin-7-O-rutinoside (1), Orientin (2) and Isoorientin (3). The methanol extract of N. ritchiana showed antioxidant activity with SC50 39.4 ± 1.06 μg/ml, while fraction IIshowed significant effect with SC50 of 6.1 ± 0.24 μg/ml in comparison with ascorbic acid (SC50 1.8 ± 0.35 μg/ml). The methanol extract and the fraction III showed a promising cytotoxic activity against selected cell lines especially the A549 with IC50 9.5 ± 1.98 μg/ml and HepG2 with IC50 8.15 ± 1.3 μg/ml, respectively. Conclusion: N. ritchiana leaf methanol extract is a new source of biologically active compounds, including flavonolignans.

Coatomer subunit beta 2 (COPB2), identified by label-free quantitative proteomics, regulates cell proliferation and apoptosis in human prostate carcinoma cells

Label-free quantitative proteomics has broad applications in the identification of differentially expressed proteins. Here, we applied this method to identify differentially expressed proteins (such as coatomer subunit beta 2 [COPB2]) and evaluated the functions and molecular mechanisms of these proteins in prostate cancer (PCA) cell proliferation. Proteins extracted from surgically resected PCA tissues and adjacent tissues of 3 patients were analyzed by label-free quantitative proteomics. The target protein was confirmed by bioinformatics and GEO dataset analyses. To investigate the role of the target protein in PCA, we used lentivirus-mediated small-interfering RNA (siRNA) to knockdown protein expression in the prostate carcinoma cell line, CWR22RV1 cells and assessed gene and protein expression by reverse transcription quantitative polymerase chain reaction and western blotting. CCK8 and colony formation assays were conducted to evaluate cell proliferation. Cell cycle distributions and apoptosis were assayed by flow cytometry. We selected the differentiation-related protein COPB2 as our target protein based on the results of label-free quantitative proteomics. High expression of COPB2 was found in PCA tissue and was related to poor overall survival based on a public dataset. Cell proliferation was significantly inhibited in COPB2-knockdown CWR22RV1 cells, as demonstrated by CCK8 and colony formation assays. Additionally, the apoptosis rate and percentage of cells in the G1 phase were increased in COPB2-knockdown cells compared with those in control cells. CDK2, CDK4, and cyclin D1 were downregulated, whereas p21 Waf1/Cip1 and p27 Kip1 were upregulated, affecting the cell cycle signaling pathway. COPB2 significantly promoted CWR22RV1 cell proliferation through the cell cycle signaling pathway. Thus, silencing of COPB2 may have therapeutic applications in PCA.

Insight on cytotoxic effects of silver nanoparticles: Alternative androgenic transactivation by adsorption with DHT

Silver nanoparticles (AgNPs) are accumulated in the male reproductive organs for a long time and cause several adverse effects in there. Up to now, there is little of information for the cytotoxic effects in male reproductive cells. In this study, the stable AgNPs with a minimal silver ion (Ag+) dissolution below concentration inducing cytotoxicity in the cell medium were exposed to the human prostate carcinoma cell line 22Rv1. Moreover particle uptake and androgen receptor (AR) transactivation were evaluated. In cell medium, AgNPs exhibited stability in an aqueous environment and minimal Ag+ release. Transmission electron microscopy (TEM) and energy dispersive spectrometer (EDS) analysis demonstrated uptake of AgNPs into cells via endocytosis, and a quantitative Ag assay showed that uptake of AgNPs was size-dependent with the majority of Ag retained in the particle form. To evaluate if the presence of AgNPs can change androgenic potentials of dihydrotestosterone (DHT, strong human androgen), we conducted an AR transactivation assay using the transgenic prostate cell line 22Rv1-MMTV-Hyg and found that AgNPs lowered androgenic transactivation of DHT, which is due to decreased bioavailability of DHT.

Effect of Levisticum officinale Hydroalcoholic Extract on DU-145 and PC-3 Prostate Cancer Cell Lines

: Levisticum officinale contains a variety of components used for its medicinal properties in traditional medicine. The previous studies showed that it had an anti-cancer effect on cell lines. The current study aims to evaluate the cytotoxic potential of Levisticum officinale hydroalcoholic extract (LOHE) on DU-145 and PC-3 human advanced prostate cancer cell lines. The anti-proliferative activity of LOHE was evaluated using the MTT assay. The results of MTT showed that the IC50 of DU-145 and PC-3 were 500 µg/mL and 400 µg/mL of LOHE, respectively, following 24 hours treatment. This study found that the hydroalcoholic extract of Levisticum officinale had beneficial cell death inducing effects on both the PC3 and DU145 human prostate carcinoma cell lines in time- and concentration-dependent fashion. Hence, LOHE may have remarkable potential for uses as new natural anticancer agent.

The 22Rv1 prostate cancer cell line carries mixed genetic ancestry: Implications for prostate cancer health disparities research using pre-clinical models.

Understanding how biological factors contribute to prostate cancer (PCa) health disparities requires mechanistic functional analysis of specific genes or pathways in pre-clinical cellular and animal models of this malignancy. The 22Rv1 human prostatic carcinoma cell line was originally derived from the parental CWR22R cell line. Although 22Rv1 has been well characterized and used in numerous mechanistic studies, no racial identifier has ever been disclosed for this cell line. In accordance with the need for racial diversity in cancer biospecimens and recent guidelines by the NIH on authentication of key biological resources, we sought to determine the ancestry of 22RV1 and authenticate previously reported racial identifications for four other PCa cell lines. We used 29 established Ancestry Informative Marker (AIM) single nucleotide polymorphisms (SNPs) to conduct DNA ancestry analysis and assign ancestral proportions to a panel of five PCa cell lines that included 22Rv1, PC3, DU145, MDA-PCa-2b, and RC-77T/E. We found that 22Rv1 carries mixed genetic ancestry. The main ancestry proportions for this cell line were 0.41 West African (AFR) and 0.42 European (EUR). In addition, we verified the previously reported racial identifications for PC3 (0.73 EUR), DU145 (0.63 EUR), MDA-PCa-2b (0.73 AFR), and RC-77T/E (0.74 AFR) cell lines. Considering the mortality disparities associated with PCa, which disproportionately affect African American men, there remains a burden on the scientific community to diversify the availability of biospecimens, including cell lines, for mechanistic studies on potential biological mediators of these disparities. This study is beneficial by identifying another PCa cell line that carries substantial AFR ancestry. This finding may also open the door to new perspectives on previously published studies using this cell line.

The Effect of a Small Molecule EZH2 Methyltransferase Inhibitor GSK126 on p53 Negative Prostate Cancer Cell Line

To quantitatively evaluate the effect of GSK126 (an inhibitor of histone methyltransferase) in inhibiting migration of prostate cancer cell line. And to investigate whether GSK126 can regulate the radiosensitivity and reduce the survival rate of p53 negative prostate cancer cell line. Human prostatic carcinoma cells (PC-3) were separated into 5 groups and incubated with GSK126 of different concentrations (0 μM, 5 μM, 10 μM, 20 μM) for 48h. The protein expression levels of EZH2 in the cells were detected by western blotting. Cell migration was examined by both wound scratch assay and transwell assay. And then the PC-3 cells were exposure to a single dose of 4Gy delivered using a 6MV photon beam of Clinac 600 CD. Cell colony assay was used to determine cell survival rates after irradiation. One group of cells without any treatments was used as the control group. All experiments were repeated 3 times, statistical analysis was performed using independent sample T-test and p value of less than 0.05 was considered to be significant. Western blotting study showed that the expression level of EZH2 in PC-3 cells decreased with increasing concentration of GSK126. Transwell assay study showed that, when the concentration of GSK126 was increased to 10μM or higher, the EZH2 expression decreased and number of cells penetrating cell membrane decreased significantly (p<0.02). Wound scratch assay study showed that, along with decreasing EZH2 expression, cell migration speed and the ability to migrate reduced (p<0.05). This is consistent with the results of transwell experiments. Results mentioned above indicated that GSK126 can regulate migration of PC-3 cells by inhibition of EZH2. Cell colony assay study showed that with the increase of GSK126 concentration, the forming rates of colonies decreased significantly after irradiation (P<0.05). It indicated that GSK126 enhanced radiosensitivity of PC-3 cells.Abstract 3402; Table 1Forming rates of colonies after irradiation of 4GyGSK126 concentrationForming rate of coloniesReference (0μM w/o radiation treatment)10 μM0.36±0.045 μM0.33±0.0510 μM0.29±0.0820 μM0.14±0.07 Open table in a new tab Using GSK126 of 10μM or higher can inhibit migration of PC-3 cells. And GSK126 in combination with radiation can effectively reduce the survival rates of PC-3 cells.

Co-assembled hybrids of proteins and carbon dots for intracellular protein delivery

Silver nanoparticles (AgNPs) are accumulated in the male reproductive organs for a long time and cause several adverse effects in there. Up to now, there is little of information for the cytotoxic effects in male reproductive cells. In this study, the stable AgNPs with a minimal silver ion (Ag+) dissolution below concentration inducing cytotoxicity in the cell medium were exposed to the human prostate carcinoma cell line 22Rv1. Moreover particle uptake and androgen receptor (AR) transactivation were evaluated. In cell medium, AgNPs exhibited stability in an aqueous environment and minimal Ag+ release. Transmission electron microscopy (TEM) and energy dispersive spectrometer (EDS) analysis demonstrated uptake of AgNPs into cells via endocytosis, and a quantitative Ag assay showed that uptake of AgNPs was size-dependent with the majority of Ag retained in the particle form. To evaluate if the presence of AgNPs can change androgenic potentials of dihydrotestosterone (DHT, strong human androgen), we conducted an AR transactivation assay using the transgenic prostate cell line 22Rv1-MMTV-Hyg and found that AgNPs lowered androgenic transactivation of DHT, which is due to decreased bioavailability of DHT.

Zinc distribution in human prostate carcinoma cell line using synchrotron X‐ray microfluorescence

Cancer is a worldwide public health problem, and its incidence in the world grew by 20% in the last decade. Zinc, an essential trace element, is involved in many cellular processes. Concerning prostate cancer, Zn could inhibit the growth of tumor cells, by inducing cell cycle arrest or apoptosis. X‐ray microfluorescence (μXRF) is an elemental analysis technique that allows mapping biologically important elements at a submillimeter scale, with high sensitivity and negligible damage to the sample. In this study, we investigated cell viability through colorimetric cytotoxicity assay (MTT) and the behavior of human prostate cell lines obtained from normal (RWPE‐1) and tumorigenic (DU145) epithelium, in three‐dimensional spheroid cultures after supplementation with ZnCl2 for 24 and 48 hr using synchrotron μXRF. The measurements were performed at the Synchrotron Light National Laboratory (Campinas, Brazil). The results of μXRF showed that Zn intensity decreased in the DU145 tumor cells independent of supplementation or treatment time, and in normal cells, the intensity increased with supplementation and treatment time. The MTT assay results showed no significant differences, which indicates that the cell viability did not change. Our results indicate that μXRF can be used as an important tool to understand the mechanism of the loss of ability to capture zinc by prostate cancer cells. Copyright © 2017 John Wiley &amp; Sons, Ltd.

Abstract 1249: Procyanidin B2-3,3′-di-O-gallate targets cancer cell metabolism in its efficacy against human prostate carcinoma cells

Abstract The metabolic profile of various cancerous tissues can be correlated with cell growth and death, specific tumor type as well as the pathological stage of tumor. In recent times, prostatic tissue metabolomics using spectroscopy methods has helped to identify and establish the metabolic profiles specific to prostate cancer (PCa) malignancy. Thus, the evaluation of the anticancer efficacy of an agent can be considered incomplete without assessing its effect on the metabolic profile of the tumor tissue. With this rationale, herein, we carried out a metabolomics study on human PCa cell lines after procyanidin B2-3,3′-di-O-gallate (B2G2) exposure, which was recently identified by us as most effective agent in grape seed extract for growth inhibition and apoptotic death of human PCa cells. Specifically, we employed quantitative high-resolution nuclear magnetic resonance spectroscopy (1H-, 13C- and 31P-NMR) to assess the metabolic profile and energy state of the B2G2-treated human PCa cell lines PC3 and C42B. This approach helped us assess global metabolic profile, including glucose metabolism, energy state, and lipid metabolism in these cells after B2G2 treatment. We also studied the time-course (4-72 hrs) of B2G2 effect on glucose uptake and lactate release in the media of B2G2-treated PCa cells. Our results indicated that there was differential effect of B2G2 on mitochondrial glucose metabolism in these cells. While glucose uptake was markedly reduced as a function of time, there was also a significant decrease in extracellular lactate export with increased B2G2 exposure-time. Importantly, B2G2 preserved citrate concentration in the PCa cells as indicated by an increase in citrate levels after treatments; citrate generation is indicative of normal secretion functions of the prostate epithelial cells. The fact that the citrate was not further utilized for cholesterol synthesis was also confirmed by a decrease in cholesterol levels in these cells, indicating decreased cholesterogenesis by B2G2. Together, these results suggest that B2G2 differentially induces metabolic alterations in various PCa cell lines, which could be associated with its effects on cell growth, proliferation and death, as well as androgen dependency (or lack of it). In in vivo study examining B2G2 effect on PCa PC3 tumor xenograft growth in athymic nu/nu mice, B2G2 treatment (dose: 5mg/kg body weight of mice; given daily as intraperitoneal injections) significantly decreased tumor volume and tumor weight by ∼61% and ∼52% (both P≤ 0.01), respectively, after 5 weeks of treatments. Taken together, these studies indicate anti-PCa efficacy of B2G2 under both in vitro and in vivo scenario and warrant further extensive studies to establish its efficacy in other PCa models. Citation Format: Chapla Agarwal, Dileep Kumar, Alpna Tyagi, Natalie Serkova, Michael Wempe, Komal Raina, Rajesh Agarwal. Procyanidin B2-3,3′-di-O-gallate targets cancer cell metabolism in its efficacy against human prostate carcinoma cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1249. doi:10.1158/1538-7445.AM2017-1249