Journal: Pharmaceuticals

Article Title: Hydrophilic and Lipophilic Carbon Dots Impart Thermosensitivity to Doxorubicin Loaded Phospholipid Liposomes

doi: 10.3390/ph19050668

Figure Lengend Snippet: Comparative cytotoxicity of either empty or DOX-loaded PEGylated DPPC liposomes (Lip), or CD (Lip-CD) and CD-C16 (Lip-CD16) containing liposomes against PC3 cells following an 1 h incubation at ( A ) 37 or ( B ) 40°C. DOX concentration was 5 or 10 μM, while the concentration of empty liposomes corresponded to the liposomal concentration in experiments at the highest DOX concentration (10 μΜ). The cytotoxicity was assessed after a 24 h post-incubation period employing the standard MTT assay. Cell survival data are the mean of at least six independent values while error bars represent one standard deviation. The statistical significance was derived by Student’s t -test following the assignment: * p < 0.05, ** p < 0.01, *** p < 0.001; ns denotes no statistical significance, p > 0.05.

Article Snippet: The human prostate cancer PC3 cell line as well as the non-cancerous human embryonic kidney HEK293 of the cell bank from the Institute of Nanoscience and Nanotechnology, NCSR Demokritos, were purchased from the American Type Culture Collection (ATCC CRL-1435TM and CRL-1573TM, respectively; Manassas, VA, USA).

Techniques: Liposomes, Incubation, Concentration Assay, MTT Assay, Standard Deviation, Derivative Assay

Journal: International Journal of Molecular Sciences

Article Title: The M1 Paradox: Pro-Tumorigenic Effect of Macrophage Cytotoxicity in Prostate Cancer

doi: 10.3390/ijms27083655

Figure Lengend Snippet: Proliferative activity and clonogenic potential of macrophage-selected prostate cancer cells. Growth kinetics and population doubling time of parental and macrophage-selected PC3 ( A ) and DU145 ( B ) cells. Clonogenic assay showing colony-forming ability of parental and PC3-res ( C ) and DU145-res ( D ) cells under low-density culture conditions. Representative images and quantitative analysis of colony number are shown. Data represent mean ± SD, n = 4.

Article Snippet: Human monocytic THP-1 cells (ATCC, Manassas, VA, USA) and human prostate cancer cells PC3 and DU145 (ATCC, USA) were cultured in RPMI 1640 medium (PanEco, Moscow, Russia) containing 10% fetal bovine serum (Biowest, Nuaille, France) and 0.1 mg/mL streptomycin/penicillin (PanEco, Moscow, Russia).

Techniques: Activity Assay, Clonogenic Assay

Journal: International Journal of Molecular Sciences

Article Title: The M1 Paradox: Pro-Tumorigenic Effect of Macrophage Cytotoxicity in Prostate Cancer

doi: 10.3390/ijms27083655

Figure Lengend Snippet: Migration capacity and gelatinase activity of macrophage-selected prostate cancer cells. ( A ) Transwell migration assay of parental and macrophage-selected PC3 and DU145 cells. Representative microphotographs of migrated cells (original magnification, ×10) and their quantitative analysis are shown. Data represent mean ± SD, n = 5. ( B ) Gelatin zymography of conditioned media from parental and macrophage-selected PC3 and DU145 cells demonstrating gelatinase activity. Samples were normalized to cell number prior to electrophoresis. Data represent mean ± SD, n = 3.

Article Snippet: Human monocytic THP-1 cells (ATCC, Manassas, VA, USA) and human prostate cancer cells PC3 and DU145 (ATCC, USA) were cultured in RPMI 1640 medium (PanEco, Moscow, Russia) containing 10% fetal bovine serum (Biowest, Nuaille, France) and 0.1 mg/mL streptomycin/penicillin (PanEco, Moscow, Russia).

Techniques: Migration, Activity Assay, Transwell Migration Assay, Zymography, Electrophoresis

Journal: International Journal of Molecular Sciences

Article Title: The M1 Paradox: Pro-Tumorigenic Effect of Macrophage Cytotoxicity in Prostate Cancer

doi: 10.3390/ijms27083655

Figure Lengend Snippet: Immunocytochemical analysis of epithelial and mesenchymal marker expression in macrophage-selected prostate cancer cells. Representative immunocytochemical staining of parental and macrophage-selected PC3 ( left panel ) and DU145 ( right panel ) cells for the epithelial markers E-cadherin and β-catenin and the mesenchymal markers smooth muscle actin (SMA) and vimentin, together with quantitative analysis of the percentage of positive cells. Staining was performed under identical experimental conditions, and representative microscopic fields are shown. Cells with visible brown DAB staining above background were scored as positive, irrespective of staining intensity. For E-cadherin and β-catenin in the PC3 model, all counted cells were positive in both parental and macrophage-selected groups (100%), and therefore no variability is visible in the corresponding graphs. Data represent mean ± SD, n = 5. Scale bar corresponds to 100 μm.

Article Snippet: Human monocytic THP-1 cells (ATCC, Manassas, VA, USA) and human prostate cancer cells PC3 and DU145 (ATCC, USA) were cultured in RPMI 1640 medium (PanEco, Moscow, Russia) containing 10% fetal bovine serum (Biowest, Nuaille, France) and 0.1 mg/mL streptomycin/penicillin (PanEco, Moscow, Russia).

Techniques: Marker, Expressing, Staining

Journal: International Journal of Molecular Sciences

Article Title: The M1 Paradox: Pro-Tumorigenic Effect of Macrophage Cytotoxicity in Prostate Cancer

doi: 10.3390/ijms27083655

Figure Lengend Snippet: Enhanced tumorigenic potential and histological features of macrophage-selected prostate cancer cells in vivo . Representative images of subcutaneous xenograft tumors formed by parental and macrophage-selected prostate cancer cells and final tumor weight measured at the experimental endpoint for PC3 ( A ) and DU145 ( B ) cells. Data represent mean ± SD, n = 5. Representative histological sections of xenograft tumors stained with hematoxylin and eosin (H&E), together with immunohistochemical staining for PU.1. H&E staining was used to assess tumor morphology, PU.1 immunostaining was used to evaluate the presence of myeloid cells within the tumor tissue ( C ). Scale bar corresponds to 100 μm.

Article Snippet: Human monocytic THP-1 cells (ATCC, Manassas, VA, USA) and human prostate cancer cells PC3 and DU145 (ATCC, USA) were cultured in RPMI 1640 medium (PanEco, Moscow, Russia) containing 10% fetal bovine serum (Biowest, Nuaille, France) and 0.1 mg/mL streptomycin/penicillin (PanEco, Moscow, Russia).

Techniques: In Vivo, Staining, Immunohistochemical staining, Immunostaining

Journal: International Journal of Molecular Sciences

Article Title: The M1 Paradox: Pro-Tumorigenic Effect of Macrophage Cytotoxicity in Prostate Cancer

doi: 10.3390/ijms27083655

Figure Lengend Snippet: Transcriptomic characterization of macrophage-selected PC3 cells. ( A ) Volcano plot showing differentially expressed genes in PC3-res cells compared with parental PC3 cells. Vertical dashed lines indicate the log 2 fold-change thresholds, and the horizontal dashed line indicates the statistical significance threshold. Genes meeting both criteria were considered differentially expressed. ( B ) Correlation between gene expression changes determined by RNA-seq and qRT-PCR for selected genes. Expression values are presented relative to parental control cells. ( C ) Gene set enrichment analysis of KEGG pathways in macrophage-selected PC3-res cells compared with parental PC3 cells. Bars indicate normalized enrichment score (NES); positive NES values represent pathways enriched among upregulated genes, whereas negative NES values represent pathways enriched among downregulated genes.

Article Snippet: Human monocytic THP-1 cells (ATCC, Manassas, VA, USA) and human prostate cancer cells PC3 and DU145 (ATCC, USA) were cultured in RPMI 1640 medium (PanEco, Moscow, Russia) containing 10% fetal bovine serum (Biowest, Nuaille, France) and 0.1 mg/mL streptomycin/penicillin (PanEco, Moscow, Russia).

Techniques: Gene Expression, RNA Sequencing, Quantitative RT-PCR, Expressing, Control

Journal: International Journal of Molecular Sciences

Article Title: The M1 Paradox: Pro-Tumorigenic Effect of Macrophage Cytotoxicity in Prostate Cancer

doi: 10.3390/ijms27083655

Figure Lengend Snippet: Selective activation of p38 MAPK, but not ERK1/2 or AKT, in macrophage-selected prostate cancer cells. Representative western blot analysis of phosphorylated p38 MAPK (Thr180/Tyr182), total p38, phosphorylated ERK1/2 (Thr202/Tyr204), total ERK1/2, phosphorylated AKT (Ser473), and total AKT in parental and macrophage-selected PC3 and DU145 cells. β-Actin was used as a loading control. Densitometric quantification of pathway activation was performed as the ratio of phosphorylated to total protein and is presented in relative units normalized to the corresponding parental control cells. Data are shown as mean ± SD, n = 3. Increased phosphorylation was observed for p38 MAPK, whereas no significant changes were detected for ERK1/2 or AKT.

Article Snippet: Human monocytic THP-1 cells (ATCC, Manassas, VA, USA) and human prostate cancer cells PC3 and DU145 (ATCC, USA) were cultured in RPMI 1640 medium (PanEco, Moscow, Russia) containing 10% fetal bovine serum (Biowest, Nuaille, France) and 0.1 mg/mL streptomycin/penicillin (PanEco, Moscow, Russia).

Techniques: Activation Assay, Western Blot, Control, Phospho-proteomics