Journal: bioRxiv

Article Title: Integrated RNA and metabolite profiling of urine liquid biopsies for prostate cancer biomarker discovery

doi: 10.1101/599514

Figure Lengend Snippet: (a) A heatmap showing expression of 37 significantly upregulated genes in cancer liquid biopsies. (b,c) Among 37 upregulated genes, 13 genes ( FH, SMARCB1, GRHPR, PHB, NACA, RPS11, RPL22, NPM1, EPCAM, TFG, HDAC6, ELK4, BRD3 ) were significantly upregulated in primary tumors (n=497) compared to normal (n=52) in TCGA data. In the heatmap, black dots next to the gene name mark the genes upregulated in primary tumor compare to normal in TCGA data. The TCGA project for PCa data is publicly available for download at https://portal.gdc.cancer.gov/projects/TCGA-PRAD . (d) These 13 genes were also tested in two prostate cancer cell lines (LNCaP and PC3), and most were overexpressed apart from NACA , which was downregulated in both cell lines; BRD3 and EPCAM , which were decreased in PC3 cells, and HDAC6 , which was downregulated in LNCaP cells. (e) Normalized Enrichment Score plot of the top 13 pathways in cancer urine. (f) Geneset Enrichment Score Plot (GSEA) of the TCA cycle (NES = 2.8, p-value = 0.0). (g) GSEA of the alanine, aspartate, and glutamate metabolism (NES = 2.42, p-value = 0.0). GSEA was conducted using GSEA software from the Broad Institute ( http://software.broadinstitute.org/gsea/index.jsp ).

Article Snippet: Prostate cancer cell lines LNCaP (ATCC ® CRL-1740TM) and PC3 (ATCC ® CRL-7934TM) were cultured in RPMI 1640 medium and Dulbecco’s Modified Eagle Medium (Thermo Fisher Scientific), respectively, supplemented with 10% FBS and penicillin/streptomycin.

Techniques: Expressing, Software

Journal: bioRxiv

Article Title: Integrated RNA and metabolite profiling of urine liquid biopsies for prostate cancer biomarker discovery

doi: 10.1101/599514

Figure Lengend Snippet: (a) GOT1 knockdown in LNCaP and PC3 prostate cancer cell lines. (b) GOT1 knockdown significantly inhibits cell viability in PC3 and LNCaP cells. (c) Invasion and (d) anchorage-independent growth in prostate cancer cell lines upon GOT1 knockdown. (e) GOT1 knockdown significantly increases ROS production in PC3 and LNCaP cells. The data from three independent experiments were expressed as mean ± SD.

Article Snippet: Prostate cancer cell lines LNCaP (ATCC ® CRL-1740TM) and PC3 (ATCC ® CRL-7934TM) were cultured in RPMI 1640 medium and Dulbecco’s Modified Eagle Medium (Thermo Fisher Scientific), respectively, supplemented with 10% FBS and penicillin/streptomycin.

Techniques: Knockdown

Journal: Marine Drugs

Article Title: Fucoidan Independently Enhances Activity in Human Immune Cells and Has a Cytostatic Effect on Prostate Cancer Cells in the Presence of Nivolumab

doi: 10.3390/md20010012

Figure Lengend Snippet: IFNγ levels in culture supernatants 72 h post-initiation of the immune/tumor co-culture, measured by HTRF-based technology on TECAN Spark microplate reader. IFNγ levels released in the supernatants of αCD3-activated PBMC at 0.05 ( b , d , f ) and 0.5 μg/mL ( c , e , g ) cultured for 72 h either alone ( a ), or with PC3 tumor cells (10:1 = PBMC:PC3 ratio), in the presence and absence of Nivolumab at 1 μg/mL), and in the presence and absence of UP ( b , c ), FV ( d , e ), and MP ( f , g ). Results are expressed as means ± SEM. * p < 0.01, ** p < 0.001 and *** p < 0.0001 for each condition when compared to the respective control. ^ p < 0.01, ^^ p < 0.001 and ^^^ p < 0.0001 for each Nivolumab-treated condition when compared to the respective Nivolumab-untreated control.

Article Snippet: Prostate cancer cell line PC3 was purchased from ATCC and used in vitro according to ATCC culture specifications.

Techniques: Co-Culture Assay, Cell Culture, Control

Journal: Marine Drugs

Article Title: Fucoidan Independently Enhances Activity in Human Immune Cells and Has a Cytostatic Effect on Prostate Cancer Cells in the Presence of Nivolumab

doi: 10.3390/md20010012

Figure Lengend Snippet: Real-time live cell monitoring of PC3 tumor cell cultures under treatment with fucoidan. PC3 tumor cells were seeded and 24 h later treated with increasing doses of—UP ( a , b ), FV ( c , d ), and MP ( e , f ). Tumor cell count ( a , c , e ) and apoptosis ( b , d , f ) were monitored and quantified over a period of ~5 days, by means of NucRed probe expression and caspase 3/7 fluorescent probe, respectively. Apoptosis is represented as an index evaluated with respect to the apoptosis events and cell number in each condition. Data were normalized and corrected to the baseline and are expressed as means ± SEM ( p < 0.001).

Article Snippet: Prostate cancer cell line PC3 was purchased from ATCC and used in vitro according to ATCC culture specifications.

Techniques: Cell Counting, Expressing

Journal: Marine Drugs

Article Title: Fucoidan Independently Enhances Activity in Human Immune Cells and Has a Cytostatic Effect on Prostate Cancer Cells in the Presence of Nivolumab

doi: 10.3390/md20010012

Figure Lengend Snippet: Real-time, live-cell monitoring of prostate PC3 tumor cell killing mediated by activated PBMCs in untreated and Nivolumab-treated conditions. Prostate PC3 tumor cells were seeded and 24 h later were co-cultured with PBMCs, in the presence of αCD3 at 0.05 or 0.5 µg/mL, in the presence and absence of Nivolumab at 1 µg/mL. Tumor cell count ( a ) and apoptosis ( b ) were monitored and quantified over a period of ~5 days, by means of NucRed probe expression and caspase 3/7 fluorescent probes, respectively, as surrogate measures of immune cell killing activity towards tumor cells. Apoptosis is represented as an index evaluated with respect to the apoptosis events and cell number in each condition. Data were normalized and corrected to the baseline and are expressed as means ± SEM ( p < 0.05).

Article Snippet: Prostate cancer cell line PC3 was purchased from ATCC and used in vitro according to ATCC culture specifications.

Techniques: Cell Culture, Cell Counting, Expressing, Activity Assay

Journal: Marine Drugs

Article Title: Fucoidan Independently Enhances Activity in Human Immune Cells and Has a Cytostatic Effect on Prostate Cancer Cells in the Presence of Nivolumab

doi: 10.3390/md20010012

Figure Lengend Snippet: Real-time live cell monitoring of prostate PC3 tumor cell killing mediated by activated PBMCs, under treatment with test compounds. Prostate PC3 tumor cells were seeded and 24 h later were co-cultured with PBMCs, in the presence of αCD3 (0.5 µg/mL), and in the presence of Nivolumab (1 µg/mL), and treated with increasing doses of each of test compounds—UP ( a , b ), FV ( c , d ), and MP ( e , f ). Tumor cell count ( a , c , e ) and apoptosis ( b , d , f ) were monitored and quantified over a period of ~5 days, by means of a NucRed probe expression and caspase 3/7 fluorescent probe, respectively, as surrogate measures of immune cell killing activity towards tumor cells. Apoptosis is represented as an index evaluated with respect to the apoptosis events and cell number in each condition. Data were normalized and corrected to the baseline and are expressed as means ± SEM ( p < 0.001).

Article Snippet: Prostate cancer cell line PC3 was purchased from ATCC and used in vitro according to ATCC culture specifications.

Techniques: Cell Culture, Cell Counting, Expressing, Activity Assay

Journal: Contrast Media & Molecular Imaging

Article Title: High Contrast PET Imaging of GRPR Expression in Prostate Cancer Using Cobalt-Labeled Bombesin Antagonist RM26

doi: 10.1155/2017/6873684

Figure Lengend Snippet: (a) In vitro specificity test of 57 Co-NOTA-PEG 2 -RM26 binding to GRPR on PC-3 (human prostate cancer) cells. Presaturation of receptors with unlabeled NOTA-PEG 2 -RM26 caused significant ( p < 0.05) reduction of cell-bound 57 Co-NOTA-PEG 2 -RM26 radioactivity. (b) Binding and internalization of 57 Co-NOTA-PEG 2 -RM26 at 37°C by PC-3 cells. Data are normalized to a maximum cell-bound radioactivity and presented as average value from 3 cell dishes ± SD. Not all error bars are visible due to the small standard deviations.

Article Snippet: PC-3 human prostate cancer cells expressing GRPR were purchased form ATCC via LGC Promochem.

Techniques: In Vitro, Binding Assay, Radioactivity

Journal: PLoS ONE

Article Title: The Response of the Prostate to Circulating Cholesterol: Activating Transcription Factor 3 (ATF3) as a Prominent Node in a Cholesterol-Sensing Network

doi: 10.1371/journal.pone.0039448

Figure Lengend Snippet: ( A ) A provisional network was generated from integration of two microarray data sets. Node color represents increases (red), no significant changes (yellow), and decreases (green) in gene expression in murine prostate tissue after cholesterol alteration as ascertained by cDNA microarray. Changes in RNA expression levels of the corresponding nodes in LNCaP cells are shown as colored node boundaries (donut shape) and the color represents increases (red), no significant change (yellow), and decreases (green) in gene expression under CDM conditions compared to control. Arrows indicate direct activation, T-shaped lines direct repression, dashed arrows indirect activation, and lines physical interaction. ( B ) Gene expression under Normo and Hyper conditions ( in vivo ). To verify in vivo microarray data obtained from SCID experiments, mRNA levels of the indicated genes were determined. GAPDH expression was used to normalize gene expression. Error bars represent SD (n = 3). ( C ) Gene expression under Control and Cholesterol-depleted conditions ( in vitro ). LNCaP cells were incubated in CDM for 0, 3 or 16 h, and mRNA levels of the indicated genes were measured by RT-PCR analysis to validate cDNA microarray data. Error bars represent SD (n = 3). * p <0.05 (Student’s t-test).

Article Snippet: LNCaP human prostate tumor cells were purchased from American Type Culture Collection (ATCC, Manassas, VA) and maintained in RPMI1640 media (Invitrogen, Carlsbad, CA) supplemented with 10% FBS and 1% Penicillin/Streptomycin at 37°C under 5% CO 2 .

Techniques: Generated, Microarray, Gene Expression, RNA Expression, Control, Activation Assay, In Vivo, Expressing, In Vitro, Incubation, Reverse Transcription Polymerase Chain Reaction

Journal: PLoS ONE

Article Title: The Response of the Prostate to Circulating Cholesterol: Activating Transcription Factor 3 (ATF3) as a Prominent Node in a Cholesterol-Sensing Network

doi: 10.1371/journal.pone.0039448

Figure Lengend Snippet: ( A ) RT-PCR analysis in vivo . ATF3 levels are reduced in all prostatic lobes from Hyper mice, compared to those from the Normo group (AP = anterior prostate; VP = ventral prostate; DLP = dorsal prostate). ( B ) Immunoblot analysis. Immunoblot of PrEC lysates showed induction of ATF3 protein by CDM (left panel) and by β-cyclodextrin (right panel). MG132, a proteasome inhibitor, also increased ATF3 expression. ( C ) Immunofluorescence analysis. Induction of ATF3 protein by CDM in LNCaP cells as shown by IF. LNCaP cells were treated with CDM for 18 h, stained with anti-ATF3 antibody and nuclei were counterstained with DAPI (left panel: ATF3; middle panel: DAPI; right panel: overlay). ( D ) RT-PCR analysis. ATF3 mRNA levels in LNCaP cells treated with CDM were normalized to levels of GAPDH. RT-PCR analysis shows induction of ATF3 mRNA levels by CDM. ( E–F ) Promoter reporter analysis. A full-length ATF3 promoter was cloned into a luciferase reporter vector and transfected into LNCaP (D) or PrEC (E). Cells were then incubated in Control and CDM medium. ATF3 promoter activity was plotted as arbitrary units (± SD) after normalization with total protein concentration.

Article Snippet: LNCaP human prostate tumor cells were purchased from American Type Culture Collection (ATCC, Manassas, VA) and maintained in RPMI1640 media (Invitrogen, Carlsbad, CA) supplemented with 10% FBS and 1% Penicillin/Streptomycin at 37°C under 5% CO 2 .

Techniques: Reverse Transcription Polymerase Chain Reaction, In Vivo, Western Blot, Expressing, Immunofluorescence, Staining, Clone Assay, Luciferase, Plasmid Preparation, Transfection, Incubation, Control, Activity Assay, Protein Concentration

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Exosomes from prostate cancer cell lines: Isolation optimisation and characterisation.

doi: 10.1016/j.biopha.2022.113093

Figure Lengend Snippet: Fig. 2. : Kinetics of exosome production by RWPE1 cells (A) and 22Rv1 cells (B) in 12-day cultivation. The graphs show concentration of cells and exosome per cell levels for both control and cancer cell lines.

Article Snippet: The human non-tumour prostatic epithelial RWPE1 cell line (ATCC CRL-11609) was cultivated in a Keratinocyte serum-free medium (Cat. No. 17005–042) supplemented with BPE (bovine pituitary extract) and EGF (epidermal growth factor) provided with the medium, and antibiotics (penicillin 100 U mL− 1, streptomycin 100 μg mL− 1).

Techniques: Concentration Assay, Control

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Exosomes from prostate cancer cell lines: Isolation optimisation and characterisation.

doi: 10.1016/j.biopha.2022.113093

Figure Lengend Snippet: Fig. 3. Exosomes produced by RWPE1 (A) and 22Rv1 (B) cell lines cultivated either on ice or at RT. Exosome per cell ratio was set to 100% at time 0 min. The effect of oxidative stress (presence of ROS) was examined by using cells RWPE1 (C) and 22Rv1 (D) exposed for 5 min to H2O2 in a buffer on ice. Two different H2O2 concentrations were used in the experiment for both cell lines. Exosome per cell ratio was set to 100% at time 0 min.

Article Snippet: The human non-tumour prostatic epithelial RWPE1 cell line (ATCC CRL-11609) was cultivated in a Keratinocyte serum-free medium (Cat. No. 17005–042) supplemented with BPE (bovine pituitary extract) and EGF (epidermal growth factor) provided with the medium, and antibiotics (penicillin 100 U mL− 1, streptomycin 100 μg mL− 1).

Techniques: Produced

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Exosomes from prostate cancer cell lines: Isolation optimisation and characterisation.

doi: 10.1016/j.biopha.2022.113093

Figure Lengend Snippet: Fig. 4. :The effect of starvation on exosome production (4 h starvation in a non-growth medium, then transfer to a growth medium) in RWPE1 and 22Rv1 cell lines. Exosome-per-cell ratio was set to 100% at time 0 min.

Article Snippet: The human non-tumour prostatic epithelial RWPE1 cell line (ATCC CRL-11609) was cultivated in a Keratinocyte serum-free medium (Cat. No. 17005–042) supplemented with BPE (bovine pituitary extract) and EGF (epidermal growth factor) provided with the medium, and antibiotics (penicillin 100 U mL− 1, streptomycin 100 μg mL− 1).

Techniques: