Prostate Adenocarcinoma Cell Line Research Articles

Synthesis of sulfanyl porphyrazines with bulky peripheral substituents – Evaluation of their photochemical properties and biological activity

Alkylation reaction of dimercaptomaleonitrile disodium salt with N-(2-bromoethyl)phthalimide in N,N-dimethylformamide, and potassium carbonate led to novel maleonitrile derivative. This compound was used in the Linstead tetramerization reaction in n-butanol in the presence of magnesium n-butoxide towards magnesium(II) sulfanyl porphyrazine with phthalimide peripheral substituents. Its demetallation and subsequent zinc(II) ion insertion led to zinc(II) sulfanyl porphyrazine derivative. All obtained compounds were thoroughly characterized using UV–vis, MS, and 1H and 13C NMR spectroscopy. The purity of macrocycles was assessed using HPLC. Novel symmetrical porphyrazines were found to generate singlet oxygen in N,N-dimethylformamide and dimethyl sulfoxide in low yields. The highest, over 4% yield of singlet oxygen generation, was noted for zinc(II) porphyrazine derivative in DMSO. In the biological study, the cytotoxicities and photocytotoxicities of symmetrical sulfanyl porphyrazines with phthalimide peripheral substituents were compared with lately obtained unsymmetrical sulfanyl tribenzoporphyrazine, 22,23-bis[3,5-bis(3,5- dimethoxybenzyloxy)benzylsulfanyl]tribenzo[b,g,l]porphyrazinato magnesium(II). The in vitro study was performed on androgen-sensitive human prostate adenocarcinoma (LNCaP) and human prostate cancer (PC3) cell lines using MTT assay. The data obtained for liposomal formulations containing sulfanyl porphyrazines with phthalimide substituents indicated their low and light-independent cytotoxicity against cancer cells. On the contrary, the sulfanyl tribenzoporphyrazine was studied in its free form and after incorporation in liposomes. In the free form it revealed moderate photocytotoxicity against both cancer cell lines with IC50 values of 7.71 μM and 8.82 μM for LNCaP and PC3 cells, respectively. Moreover, the liposomal DOTAP : POPC formulation containing sulfanyl tribenzoporphyrazine revealed over 10-fold higher effectiveness reaching the IC50 up to 0.52 μM against LNCaP cancer cells in comparison to its free form. On the contrary, liposomal PG formulation of sulfanyl tribenzoporphyrazine was found to be inactive on the human cancer cells.

Synthesis, molecular modeling, in vivo study and anticancer activity against prostate cancer of (+) (S)-naproxen derivatives

In this study, (S)-naproxen thiosemicarbazides (3a-d), 1,2,4-triazoles (4a-c), triazole-thioether hybride compounds (5a-p) were synthesized and their structures (3a, 3d, 4a and 5a-p) were confirmed by FT-IR, 1H NMR,13C NMR, HR-Mass spectra and elemental analysis. These compounds are designed to inhibit methionine amino peptidase-2 (MetAP2) enzyme in prostate cancer. These compounds (3d, 5a-p) evaluated against androgen-independent prostate adenocarcinoma (PC-3, DU-145) and androgen-dependent prostate adenocarcinoma (LNCaP) cell lines by using MTS method. Compounds 5a, 5b, 5d and 5e showed 14.2, 5.8, 10.8 and 8.4μM anticancer activity against PC-3cell lines, compounds 5e, 5g and 5n presented anticancer activity against DU-145cell lines 18.8, 12.25 and 10.2μM, and compounds 5g, 5m and 5n exhibited anticancer activity against LNCaP cell lines 12.25, 22.76 and 2.21μM, respectively. Consequently, of these results, compounds 5e and 5n showed the highest activities against androgen dependent and independent prostate cancer cell lines, so these compounds could be potent small molecules against prostate cancer. Furthermore, mitogen-activated protein kinase (MAPK) pathway activation, AKT (protein kinase B) phosphorylation and androgen receptor activation of compound 5n (SGK636) were investigated in LNCaP cells by using Western blot method. Compound 5n (SGK636) was also tested against mRNA expression analysis of the Bax, Bcl-2, Caspase 3, Caspase 9 by using real-time PCR analysis. Compound 5n was given to nude male mice with cancer in comparison to the control group. Compound 5n was found to reverse the malignant phenotype in the nude male mice, whereas the prostate cancer progressed in the control group. Analysis of some blood parameters in the study showed that they were within the normal values with respect to the control. The blood values of the animals treated according to the control group also exhibited compliance with the blood limit values. Molecular docking and dynamics simulation of compound 5n binding to Methionine Aminopeptidase 2 (MetAP2) enzyme rationalized its potential activity. In addition, inhibition assay MetAP2 enzyme of compound 5n was evaluated. Taken together, we suggest compound 5n to be a potential candidate for prostate cancer therapy.

Transcript Levels of Aldo-Keto Reductase Family 1 Subfamily C (AKR1C) Are Increased in Prostate Tissue of Patients with Type 2 Diabetes.

Aldo-keto reductase family 1 (AKR1) enzymes play a crucial role in diabetic complications. Since type 2 diabetes (T2D) is associated with cancer progression, we investigated the impact of diabetes on AKR1 gene expression in the context of prostate cancer (PCa) development. In this study, we analyzed benign (BEN) prostate and PCa tissue of patients with and without T2D. Furthermore, to replicate hyperglycemia in vitro, we treated the prostate adenocarcinoma cell line PC3 with increasing glucose concentrations. Gene expression was quantified using real-time qPCR. In the prostate tissue of patients with T2D, AKR1C1 and AKR1C2 transcripts were higher compared to samples of patients without diabetes. In PC3 cells, high glucose treatment induced the gene expression levels of AKR1C1, C2, and C3. Furthermore, both in human tissue and in PC3 cells, the transcript levels of AKR1C1, C2, and C3 showed positive associations with oncogenes, which are involved in proliferation processes and HIF1α and NFκB pathways. These results indicate that in the prostate glands of patients with T2D, hyperglycemia could play a pivotal role by inducing the expression of AKR1C1, C2, and C3. The higher transcript level of AKR1C was furthermore associated with upregulated HIF1α and NFκB pathways, which are major drivers of PCa carcinogenesis.

Synthesis and Anticancer Activity of New Pyridine-Thiophene and Pyridine-Furan Hybrid Compounds, Their Sugar Hydrazone, and Glycosyl Derivatives

New sugar hydrazones and their derived oxadiazolyl acyclic nucleoside analogs in addition to the derived thioglycosides incorporating pyridine, furan or thiophene have been synthesized via a multi-component reaction (MCR). The acetylated derivatives of hydrazones and the derived deprotected thioglycosides of the prepared acetylated analogs have been also synthesized. Anticancer activity of the products has been tested against adenocarcinomic human alveolar basal epithelial (A549), human prostatic adenocarcinoma (PC3) and human colorectal carcinoma (HCT116) cell lines in addition to their effect on human normal retinal pigmented epithelial cell line (RPE1) using the MTT assay. Three products: acid hydrazide, sugar hydrazine and glycoside are characterized by high activity against three cancer cell lines.

Protective effects of combination of Stauntonia hexaphylla and Cornus officinalis on testosterone-induced benign prostatic hyperplasia through inhibition of 5α- reductase type 2 and induced cell apoptosis

Benign prostatic hyperplasia (BPH) is a progressive pathological condition associated with proliferation of prostatic tissues, prostate enlargement, and lower-urinary tract symptoms. However, the mechanism underlying the pathogenesis of BPH is unclear. The aim of this study was to investigate the protective effects of a combination of Stauntonia hexaphylla and Cornus officinalis (SC extract) on a testosterone propionate (TP)-induced BPH model. The effect of SC extract was examined in a TP-induced human prostate adenocarcinoma cell line. Male Sprague-Dawley rats were randomly divided into 5 groups (n = 6) for in vivo experiments. To induce BPH, all rats, except those in the control group, were administered daily with subcutaneous injections of TP (5 mg/kg) and orally treated with appropriate phosphate buffered saline/drugs (finasteride/saw palmetto/SC extract) for 4 consecutive weeks. SC extract significantly downregulated the androgen receptor (AR), prostate specific antigen (PSA), and 5α-reductase type 2 in TP-induced BPH in vitro. In in vivo experiments, SC extract significantly reduced prostate weight, size, serum testosterone, and dihydrotestosterone (DHT) levels. Histologically, SC extract markedly recovered TP-induced abnormalities and reduced prostatic hyperplasia, thereby improving the histo-architecture of TP-induced BPH rats. SC extract also significantly downregulated AR and PSA expression, as assayed using immunoblotting. Immunostaining revealed that SC extract markedly reduced the 5α-reductase type 2 and significantly downregulated the expression of proliferating cell nuclear antigen. In addition, immunoblotting of B-cell lymphoma 2 (Bcl-2) family proteins indicated that SC extract significantly downregulated anti-apoptotic Bcl-2 and markedly upregulated pro-apoptotic B cell lymphoma-associated X (Bax) expression. Furthermore, SC treatment significantly decreased the Bcl-2/Bax ratio, indicating induced prostate cell apoptosis in TP-induced BPH rats. Thus, our findings demonstrated that SC extract protects against BPH by inhibiting 5α-reductase type 2 and inducing prostate cell apoptosis. Therefore, SC extract might be useful in the clinical treatment of BPH.

Proteins from Lignosus tigris with selective apoptotic cytotoxicity towards MCF7 cell line and suppresses MCF7-xenograft tumor growth.

BackgroundLignosus tigris, a recently discovered species of the unique Lignosus family, has been traditionally used by the indigenous communities in Peninsular Malaysia to treat various ailments and as an alternative medicine for cancer treatment. The L. tigris cultivar sclerotia (Ligno TG-K) was found to contain numerous bioactive compounds with beneficial biomedicinal properties and the sclerotial extract exhibited potent antioxidant activity. However, the anticancer property of the Ligno TG-K including in vitro and in vivo antitumor effects as well as its anticancer active compounds and the mechanisms has yet to be investigated.MethodsThe cytotoxicity of the Ligno TG-K against human breast (MCF7), prostate (PC3) and lung (A549) adenocarcinoma cell lines was evaluated using MTT cytotoxicity assay. The cytotoxic mechanisms of the active high molecular weight proteins (HMWp) fraction were investigated through detection of caspases activity and apoptotic-related proteins expression by Western blotting. The in vivo antitumor activity of the isolated HMWp was examined using MCF7 mouse xenograft model. Shotgun LC-MS/MS analysis was performed to identify the proteins in the HMWp.Results and DiscussionCold water extract of the sclerotia inhibited proliferation of MCF7, A549 and PC3 cells with IC50 ranged from 28.9 to 95.0 µg/mL. Bioassay guided fractionation of the extract revealed that HMWp exhibited selective cytotoxicity against MCF7 cells via induction of cellular apoptosis by the activation of extrinsic and intrinsic signaling pathways. HMWp activated expression of caspase-8 and -9 enzymes, and pro-apoptotic Bax protein whilst inhibiting expression of tumor survivor protein, Bcl-2. HMWp induced tumor-cell apoptosis and suppressed growth of tumor in MCF-7 xenograft mice. Lectins, serine proteases, RNase Gf29 and a 230NA deoxyribonuclease are the major cytotoxic proteins that accounted for 55.93% of the HMWp.ConclusionThe findings from this study provided scientific evidences to support the traditional use of the L. tigris sclerotia for treatment of breast cancer. Several cytotoxic proteins with high abundance have been identified in the HMWp of the sclerotial extract and these proteins have potential to be developed into new anticancer agents or as adjunct cancer therapy.

CXCL12γ induces human prostate and mammary gland development.

Epithelial stem cells (ESCs) demonstrate a capacity to maintain normal tissues homeostasis and ESCs with a deregulated behavior can contribute to cancer development. The ability to reprogram normal tissue epithelial cells into prostate or mammary stem-like cells holds great promise to help understand cell of origin and lineage plasticity in prostate and breast cancers in addition to understanding normal gland development. We previously showed that an intracellular chemokine, CXCL12γ induced cancer stem cells and neuroendocrine characteristics in both prostate and breast adenocarcinoma cell lines. However, its role in normal prostate or mammary epithelial cell fate and development remains unknown. Therefore, we sought to elucidate the functional role of CXCL12γ in the regulation of ESCs and tissue development. Prostate epithelial cells (PNT2) or mammary epithelial cells (MCF10A) with overexpressed CXCL12γ was characterized by quantitative real-time polymerase chain reaction, Western blots, and immunofluorescence for lineage marker expression, and fluorescence activated cell sorting analyses and sphere formation assays to examine stem cell surface phenotype and function. Xenotransplantation animal models were used to evaluate gland or acini formation in vivo. Overexpression of CXCL12γ promotes the reprogramming of cells with a differentiated luminal phenotype to a nonluminal phenotype in both prostate (PNT2) and mammary (MCF10A) epithelial cells. The CXCL12γ-mediated nonluminal type cells results in an increase of epithelial stem-like phenotype including the subpopulation of EPCAMLo /CD49fHi /CD24Lo /CD44Hi cells capable of sphere formation. Critically, overexpression of CXCL12γ promotes the generation of robust gland-like structures from both prostate and mammary epithelial cells in in vivo xenograft animal models. CXCL12γ supports the reprogramming of epithelial cells into nonluminal cell-derived stem cells, which facilitates gland development.

Chemical compositions of essential oils of Amomum verum and Cinnamomum parthenoxylon and their in vitro biological properties

Introduction: In eastern Thailand, Amomum verum and Cinnamomum parthenoxylon are native plants used by local communities for their medical and culinary properties. This study determined the chemical composition and biological activities of the essential oils from A. verum shoots (AVSEO) and C. parthenoxylon wood (CPW-EO). Methods: Essential oils were extracted using hydro-distillation and analyzed using gas chromatography-mass spectroscopy (GC-MS). The antimicrobial activity was evaluated using the disc diffusion method and broth microdilution assay. The cytotoxic activity of the essential oils was assessed against the human prostate adenocarcinoma (DU145) cell line using 3-(4,5-dimethylthiazol2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The antioxidant activity of the essential oils was determined using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis-(3-ethylbenzothiazoline6-sulfonic acid) (ABTS) radical scavenging assays. The expression of antioxidant genes in the DU145 cells was evaluated using quantitative real-time polymerase chain reaction (qRT-PCR). Results: 1,8-Cineole was the main component in AVS-EO and CPW-EO with 84.38, and 45.65 %, respectively. AVS-EO had stronger antimicrobial activity than CPW-EO. The lowest minimum inhibitory concentration (MIC) and minimum microbicidal concentration (MMC) values of AVSEO against Candida albicans were 0.3125 and 2.5 mg/mL, respectively. Both essential oils had timedependent and dose-dependent cytotoxic effects on the DU145 human prostate adenocarcinoma cells. CPW-EO had high antioxidant activity toward DPPH and ABTS radicals with IC50 values of 4.528±0.233 and 0.045±0.007 mg/mL, respectively. The two essential oils up-regulated glutathione peroxidase (GPx) and glutathione reductase (GRx) mRNA expression in the oxidative stress response of DU145 cells. Conclusion: AVS-EO and CPW-EO might be added as natural ingredients in food or dietary supplement products for the benefit of microbial and prostate cancer inhibition.

The n-10 Fatty Acids Family in the Lipidome of Human Prostatic Adenocarcinoma Cell Membranes and Extracellular Vesicles.

A new pathway leading to the n-10 fatty acid series has been recently evidenced, starting from sapienic acid, a monounsaturated fatty acid (MUFA) resulting from the transformation of palmitic acid by delta-6 desaturase. Sapienic acid has attracted attention as a novel marker of cancer cell plasticity. Here, we analyzed fatty acids, including the n-10 fatty acid contents, and for the first time, compared cell membranes and the corresponding extracellular vesicles (EV) of two human prostatic adenocarcinoma cell lines of different aggressiveness (PC3 and LNCaP). The n-10 components were 9–13% of the total fatty acids in both cancer cell lines and EVs, with total MUFA levels significantly higher in EVs of the most aggressive cell type (PC3). High sapienic/palmitoleic ratios indicated the preference for delta-6 versus delta-9 desaturase enzymatic activity in these cell lines. The expressions analysis of enzymes involved in desaturation and elongation by qRT-PCR showed a higher desaturase activity in PC3 and a higher elongase activity toward polyunsaturated fatty acids than toward saturated fatty acids, compared to LNCaP cells. Our results improve the present knowledge in cancer fatty acid metabolism and lipid phenotypes, highlighting EV lipidomics to monitor positional fatty acid isomer profiles and MUFA levels in cancer.

Identifying the key genes and microRNAs in prostate cancer bone metastasis by bioinformatics analysis.

Prostate adenocarcinoma (PCa) is the most common cause of death due to malignancy among men, and bone metastasis is the leading cause of mortality in patients with PCa. Therefore, identifying the causes and molecular mechanism of bone metastasis is important for early detection, diagnosis and personalized therapy. In this study, we systematically analyzed molecular correlates of bone metastasis by bioinformatics analysis. A total of 12 differentially expressed microRNAs (miRNAs) and 102 differentially expressed genes were identified. Five miRNAs had prognostic significance in biochemical recurrence‐free survival (miR‐636, miR‐491‐5p, miR‐199b‐5p, miR‐199b‐3p, miR‐28‐3p). The differentially expressed genes were significantly enriched in extracellular matrix, cell‐substrate adhesion, collagen and integrin. Seven hub genes (VCAN, COL3A1, COL1A1, APOE, COL1A2, SDC1, THY1) with worse biochemical recurrence‐free survival and one hub gene (MMP9) with worse overall survival were detected. miR‐636, a novel oncogene, was found to be up‐regulated in bone metastatic PCa tissues and also predominately up‐regulated in human PCa cell lines. miR‐636 promoted cellular invasion and migration, and may promote bone metastasis via targeting MBNL2, TNS1 and STAB1. In conclusion, we have successfully defined molecular signatures of bone metastasis in PCa.

Characterization of six canine prostate adenocarcinoma and three transitional cell carcinoma cell lines derived from primary tumor tissues as well as metastasis.

Canine prostate adenocarcinoma (PAC) and transitional cell carcinoma (TCC) of prostate and urinary bladder are highly invasive and metastatic tumors of closely neighbored organs. Cell lines are valuable tools to investigate tumor mechanisms and therapeutic approaches in vitro. PAC in dogs is infrequent, difficult to differentiate from TCC and usually characterized by poor prognosis, enhancing the value of the few available cell lines. However, as cell lines adapt to culturing conditions, a thorough characterization, ideally compared to original tissue, is indispensable. Herein, six canine PAC cell lines and three TCC cell lines were profiled by immunophenotype in comparison to respective original tumor tissues. Three of the six PAC cell lines were derived from primary tumor and metastases of the same patient. Further, two of the three TCC cell lines were derived from TCCs invading into or originating from the prostate. Cell biologic parameters as doubling times and chemoresistances to commonly used drugs in cancer treatment (doxorubicin, carboplatin and meloxicam) were assessed. All cell lines were immunohistochemically close to the respective original tissue. Compared to primary tumor cell lines, metastasis-derived cell lines were more chemoresistant to doxorubicin, but equally susceptive to carboplatin treatment. Two cell lines were multiresistant. COX-2 enzyme activity was demonstrated in all cell lines. However, meloxicam inhibited prostaglandin E2 production in only seven of nine cell lines and did neither influence metabolic activity, nor proliferation. The characterized nine cell lines represent excellent tools to investigate PAC as well as TCC in prostate and urinary bladder of the dog. Furthermore, the profiled paired cell lines from PAC primary tumor and metastasis provide the unique opportunity to investigate metastasis-associated changes PAC cells undergo in tumor progression. The combination of nine differently chemoresistant PAC and TCC cell lines resembles the heterogeneity of canine lower urinary tract cancer.

Molecular and Cellular Risk Assessment of Healthy Human Cells and Cancer Human Cells Exposed to Nanoparticles.

Nanodrugs have in recent years been a subject of great debate. In 2017 alone, almost 50 nanodrugs were approved for clinical use worldwide. Despite the advantages related to nanodrugs/nanomedicine, there is still a lack of information regarding the biological safety, as the real behavior of these nanodrugs in the body. In order to better understand these aspects, in this study, we evaluated the effect of polylactic acid (PLA) nanoparticles (NPs) and magnetic core mesoporous silica nanoparticles (MMSN), of 1000 nm and 50 nm, respectively, on human cells. In this direction we evaluated the cell cycle, cytochemistry, proliferation and tubulogenesis on tumor cells lines: from melanoma (MV3), breast cancer (MCF-7, MDA-MB-213), glioma (U373MG), prostate (PC3), gastric (AGS) and colon adenocarcinoma (HT-29) and non-tumor cell lines: from human melanocyte (NGM), fibroblast (FGH) and endothelial (HUVEC), respectively. The data showed that an acute exposure to both, polymeric nanoparticles or MMSN, did not show any relevant toxic effects on neither tumor cells nor non-tumor cells, suggesting that although nanodrugs may present unrevealed aspects, under acute exposition to human cells they are harmless.

Non-porous phosphonated ionic silica nanospheres as nanocarriers for efficient intracellular delivery of doxorubicin

A novel nanoscale drug delivery system based on monodispersed non-porous ionic silica nanospheres (NISNs) decorated with phosphonated active sides homogeneously distributed all over their surface, was developed. Doxorubicin (DOX), a well-known antitumor drug, was successfully loaded on the surface of the silica nanoparticles via electrostatic interactions. The final drug vehicle possesses excellent solubility, while enhancing significantly the efficacy of the drug. The administration of DOX-loaded NISNs against two aggressive DOX-resistant human prostate adenocarcinoma cell lines DU145 and PC3 leads to increased medicinal efficacy with extremely low DOX concentrations (0.1 μM) that could significantly reduce DOX side effects. In addition, NISNs was found to be non-toxic. The efficient cellular uptake of NISNs_DOX was confirmed by flow cytometry analysis and visualized by confocal microscopy. The translocation of DOX inside cells drastically changed, when DOX was bound to NISNs nanoparticles. Specifically, DOX loaded to NISNs nanoparticles is preferentially localized in the cytosol and significant efficacy was observed due to slow controlled release of DOX to the nucleus. The results reported in this work strongly support the potential utilization of NISNs derivatives as non-toxic nanocarriers for high loading efficiency and intracellular delivery of therapeutic drugs.

PARP Inhibition Suppresses GR-MYCN-CDK5-RB1-E2F1 Signaling and Neuroendocrine Differentiation in Castration-Resistant Prostate Cancer.

In this study, we addressed the underlying mechanisms for the association between enzalutamide (ENZ) treatment and neuroendocrine prostate cancer (NEPC), and the critical involvement of MYCN, and loss of RB1 function in neuroendocrine differentiation (NED) of prostatic epithelial cells, and the development of NEPC. We further sought to determine whether PARP inhibition could suppress NEPC, and to identify molecular determinants of this therapeutic activity. We used a novel prostate cancer patient-derived xenograft (PDX) treatment model, prostatic adenocarcinoma and NEPC cell lines, an NEPC organoid line, and NEPC xenograft models to address the mechanistic basis of ENZ-induced NED, and to analyze suppression of NED and NEPC growth by PARP inhibition. We identified an ENZ treatment-associated glucocorticoid receptor (GR)-MYCN-CDK5-RB1-E2F1 signaling pathway that drives NED in prostatic adenocarcinoma PDX and cell line models. Mechanistically, long-term ENZ treatment transcriptionally upregulates signaling of the GR-MYCN axis, leading to CDK5R1 and CDK5R2 upregulation, Rb1 phosphorylation, and N-Myc-mediated and E2F1-mediated NED gene expression. Importantly, olaparib (OLA) or talazoparib (TALA) suppressed these activities, and the combination of OLA and dinaciclib (DINA), an inhibitor of CDK2 and CDK5, which also inhibits Rb1 phosphorylation, suppressed NED and significantly improved therapeutic efficiency in NEPC cells in vitro and in NEPC tumors in vivo. The results of our study indicate an important role of GR-MYCN-CDK5R1/2-RB1-NED signaling in ENZ-induced and PARP inhibitor-suppressed NEPC. We also demonstrated efficacy for OLA+DINA combination therapy in NEPC xenograft models.

Synthesis of silver nanoparticles (AgNPs) from leaf extract of Salvia miltiorrhiza and its anticancer potential in human prostate cancer LNCaP cell lines

Biosynthesis of silver nanoparticles (AgNPs) from the medicinal plants has been considered as a remarkable approach of several therapeutic innovations and successful drug delivery. Silver nanoparticles were biosynthesized with Salvia miltiorrhiza, Chinese medicinal herb and assessed for its anticarcinogenic property. Synthesis of AgNPs was characterized by several studies such as UV-absorbance and it shows peak values in the range of 425–445 nm. The sizes of the nanoparticles are confirmed by dynamic light scattering analysis and it shows 100 nm. Furthermore, transmission electron microscopy (TEM) and energy dispersive X-ray analysis (EDX) was to confirm the shape and Ag particles are present in the synthesized materials. FTIR analysis to find out the active biomolecules located in the surface of the synthesized particles. This AgNPs from S. miltiorrhiza inhibits the growth of Bacillus subtillis, Staphylococcus aureus, Escherichia coli, and Klebsiella pneumonia. Furthermore, the anticancer potential of AgNPs is examined in prostate adenocarcinoma (LNCaP) cell lines. In this study, we found the AgNPs effectively induces cytotoxicity, ROS and apoptosis by modulation of intrinsic apoptoic Bcl2, Bclxl, Bax and Caspase 3 protein expressions in LNCap cell lines. Based on the study, synthesis of AgNPs from S. miltiorrhiza shows eco-friendly and it exhibits antimicrobial and anticarcinogenic effects.

Abstract 3017: Role of long noncoding RNA H19 in driving enzalutamide resistant neuroendocrine prostate cancer

Abstract INTRODUCTION: Approximately one-fourth of the prostate cancer (PCa) patients relapsing androgen deprivation therapy (ADT) have been shown to develop neuroendocrine prostate cancer (NEPC). One of the primary mechanisms attributed to this resistance is the lineage switching of epithelial cells to neuroendocrine (NE) phenotype making them independent of androgen receptor (AR) pathways for survival. Molecular basis essential to this lineage switch requires further elucidation. Thus, we aim to understand key mechanistic pathways that drive the development and maintenance of NEPC, and to investigate if the inhibition of pathway can re-sensitize to ADT in NEPC. METHODS: Bioinformatics analysis was performed with existing databases to compare prostate adenocarcinoma (PCA) vs NEPC. Molecular features of NEPC have been shown to depict RB1 loss in 70-90% and TP53 loss in 56-67% of the cases. Thus, we established prostate organoid cultures from P53fl/fl/Rbfl/fl mouse and infected them with Cre-lentivirus for combined loss of TP53/Rb1. Various PCA and NEPC cell lines and patient derived organoids were used to study the role of the long non-coding RNA H19 in NEPC. Organoid growth were assessed with Incucyte and invasive capacity was measured by Matrigel invasion assay. RESULTS: Bioinformatic analysis of exisiting human PCa data sets demonstrate that H19 is one of the most highly expressed genes in NEPC and parallels the expression of NE markers. Indeed, we confirmed that H19 expression was high in various NEPC cell lines and NE patient derived organoids. H19 was also found to be markedly induced in mouse prostate organoids and LNCaP cells after TP53/Rb1 loss. Importantly, overexpression of H19 induced NE genes while suppressing the genes involved in AR signaling in LNCaP and primary PCa patient derived organoids and inversely, knockdown (KD) of H19 in LNCaP and mouse prostate organoids with TP53/Rb1 loss reduced the expression of NE gene. Interestingly, the KD of H19 in these cells re-induced the luminal phenotype, consequently restoring their sensitivity to ADT. These findings strongly implicate H19 in driving the NE lineage switch. The KD of H19 in these organoids led to a regression of tumor growth and invasiveness, suggesting H19 is required in malignant states. H19 KD decreases stem cell genes (SOX2, OCT4) via methylation of their promoters, and levels of EZH2, which methylates 'Lys-9' (H3K9me) and 'Lys-27' (H3K27me) of histone H3 and represses transcription of the target genes. H3K27me3 levels, which are high in NEPC, was reduced in the H19 KD, suggesting H19 works in an epigenetic regulation to induce NEPC phenotype. CONCLUSION: These data highlight H19 as a novel regulator in controlling the lineage switch to NEPC, tumor growth, and the sensitivity to hormone blockade, suggesting that H19 regulation will be important for developing treatments specifically aimed for this highly aggressive form of prostate cancer. Citation Format: Neha Singh, Virginie Olive, Ritu Pandey, Jin Song, Jeremiah Bearrs, Sathish K. Padi, Koichi Okumura, Andrew S. Kraft. Role of long noncoding RNA H19 in driving enzalutamide resistant neuroendocrine prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3017.

Abstract 4285: p66Shc/ROS enhances the progression of androgen-sensitive towards castration-resistant prostate cancer cells

Abstract Purpose and Background: Prostate cancer (PCa) is the second leading cause of cancer-related death in United States men. Androgen deprivation therapy is the standard-of-care treatment for metastatic PCa; most patients eventually relapse and develop castration-resistant (CR) tumors that currently have no effective treatment. Thus, a useful cell model for analysis of the molecular mechanism of PCa progression is required for developing targeted therapies for CR PCa. In this study, we established a PCa cell progressive model in three independent cell lines, of which androgen-independent (AI) cells were derived from respective androgen-sensitive (AS) cells, recapitulating clinical PCa progression. Experimental Methods: AS and AI human prostate adenocarcinoma cell lines LNCaP, MDA PCa2b and VCaP were utilized in this study to demonstrate the molecular and signaling alterations seen in CR PCa. Stable p66Shc cDNA transfected subclones were established from LNCaP-AS cells. The tumorigenicity of AS and AI cells as well as AS PCa cells treated with H2O2 and NAC were evaluated via trypan blue exclusion, transwell migration, clonogenic assays and xenograft mouse models. The signaling profiles including phosphoprotein microarray and immunoblotting were conducted. Results: AI PCa cells have enhanced tumorigenicity, recapitulating the clinical CR phenotype, including AR expression, proliferation and tumorigenicity under androgen-deprived conditions. Further, AI cells exhibit increased ROS levels as well as enhanced signaling of proliferation and survival pathways. We further identified oxidase p66Shc as one of the potential sources of the ROS-mediated phenotypic and cell signaling alterations in AI PCa cells. LNCaP-AI cells and p66Shc subclones have a greater oxidative environment compared to LNCaP-AS cells. Increased ROS via H2O2 enhanced AS cell growth and migration, which was counteracted by antioxidant NAC. Treatment of LNCaP-AS cells with H2O2 resulted in a similar signaling profile to that of LNCaP-AI or p66Shc subclone cells. Further, the ROS-driven alterations of p66Shc subclone cell signaling can be mitigated via p66Shc knockdown or inactive p66Shc mutant. Moreover, LNCaP-AI cells and p66Shc subclones, but not LNCaP-AS cells, develop xenograft tumors with metastatic nodules. Molecular profiling showed that alterations of signaling in LNCaP-AI cells and p66Shc subclones attributed to p66Shc/ROS. Conclusions: We report the establishment of a PCa cell progression model in three commonly used PCa cell lines that replicates clinical PCa progression from the AS to the AI/CR phenotype. Altogether, the data shows ROS produced by p66Shc promotes PCa tumorigenicity and progression to the CR phenotype. Further characterization of the PCa progressive model will aid in the understanding of advanced PCa progression to help in treatment of this lethal disease. Citation Format: Dannah Miller, Matthew Ingersoll, Arpita Chatterjee, Rebecca Oberley-Deegan, Ming-Fong Lin. p66Shc/ROS enhances the progression of androgen-sensitive towards castration-resistant prostate cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4285.

Multi-Omics Analyses Detail Metabolic Reprogramming in Lipids, Carnitines, and Use of Glycolytic Intermediates between Prostate Small Cell Neuroendocrine Carcinoma and Prostate Adenocarcinoma.

As the most common cancer in men, prostate cancer is molecularly heterogeneous. Contributing to this heterogeneity are the poorly understood metabolic adaptations of the two main types of prostate cancer, i.e., adenocarcinoma and small cell neuroendocrine carcinoma (SCNC), the latter being more aggressive and lethal. Using transcriptomics, untargeted metabolomics and lipidomics profiling on LASCPC-01 (prostate SCNC) and LNCAP (prostate adenocarcinoma) cell lines, we found significant differences in the cellular phenotypes of the two cell lines. Gene set enrichment analysis on the transcriptomics data showed 62 gene sets were upregulated in LASCPC-01, while 112 gene sets were upregulated in LNCAP. ChemRICH analysis on metabolomics and lipidomics data revealed a total of 25 metabolite clusters were significantly different. LASCPC-01 exhibited a higher glycolytic activity and lower levels of triglycerides, while the LNCAP cell line showed increases in one-carbon metabolism as an exit route of glycolytic intermediates and a decrease in carnitine, a mitochondrial lipid transporter. Our findings pinpoint differences in prostate neuroendocrine carcinoma versus prostate adenocarcinoma that could lead to new therapeutic targets in each type.

MicroRNA-205 and microRNA-338-3p Reduces Cell Apoptosis in Prostate Carcinoma Tissue and LNCaP Prostate Carcinoma Cells by Directly Targeting the B-Cell Lymphoma 2 (Bcl-2) Gene.

BackgroundThe inhibitor of apoptosis, B-cell lymphoma 2 (Bcl-2), is encoded by the BCL2 gene. Previous studies have shown that microRNAs are downregulated in prostate cancer. This study aimed to investigate the role of microRNA-205 and microRNA-338-3p and cell apoptosis in prostate carcinoma tissue and the LNCaP human prostate adenocarcinoma cell line by directly targeting the BCL2 gene and Bcl-2 protein expression.Material/MethodsBioinformatics methods predicted the target genes of miR-205 and miR-338-3p, which were validated by a luciferase assay. Immunohistochemistry was used to detect Bcl-2 protein expression in 30 samples of prostate carcinoma tissue and 30 matched samples of normal prostate. The normal prostate epithelial cell line, RWPE-1, and LNCaP human prostate adenocarcinoma cells studied in vitro. BCL2 mRNA expression and Bcl-2 protein expression were determined by quantitative polymerase chain reaction (q-PCR) and Western blot, respectively. Cell apoptosis was measured by flow cytometry using annexin V, fluorescein isothiocyanate, and phycoerythrin (annexin V-FITC/PE).ResultsTargetScan Human 7.2 predicted that the structures of miR-205 and miR-338-3p had a binding site on the proto-oncogene, BCL2, which was verified by a luciferase assay. The expression of miR-205 and miR-338-3p were significantly downregulated in prostate carcinoma tissues and LNCaP cells when compared with normal controls. BCL2 expression was significantly inhibited by overexpression of miR-205 and miR-338-3p in LNCaP cells.ConclusionsThe results of this study showed that miR-205 and miR-338-3p downregulated the expression of the BCL2 gene and decreased apoptosis in prostate carcinoma.

Effect of Exposure to Quran Recitation on Cell Viability, Cell Migration, and BCL2L12 Gene Expression of Human Prostate Adenocarcinoma Cell Line in Culture

Effect of Exposure to Quran Recitation on Cell Viability, Cell Migration, and BCL2L12 Gene Expression of Human Prostate Adenocarcinoma Cell Line in Culture