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ATCC human pc3
Human Pc3, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 14405 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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human pc3 - by Bioz Stars, 2026-07

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The metabolic component secreted by senescent HPFs and HOFs is involved in the acquisition of a pro-invasive phenotype in prostate and ovarian cancer cells. ( A ) Quantification of SA-β-Gal staining of human prostate fibroblasts (HPFs) and human ovarian fibroblasts (HOFs) treated for 24 h with 5 nM Docetaxel (DTX) and 20 μM cisplatin (CPT), respectively, and then cultured in drug-free medium for an additional 6 days. For each condition, images were taken from five randomly selected fields, and both the total number of cells and the number of blue (SA-β-Gal positive) cells were counted. Bar graphs represent the average ratio of positive cells to the total cell count. Representative images of the stained cells are shown below the bar graphs (magnification 20×, scale bar 100 μm). ( B ) Representative immunoblots of p16 and p21 protein levels in DTX-treated HPFs and CPT-treated HOFs. β-actin was used as loading control. ( C ) Invasion assay: PC3 and SKOV3 cells were incubated with CM CTRL and CM SEN conditioned media (CM) from senescent and non-senescent (CTRL) fibroblasts for 72 h and then seeded in Boyden chambers. Cells were allowed to invade for 16 h. Representative images of the filters are shown below the bar graphs (magnification 20×, scale bar 100 μm). ( D ) mRNA expression level of EMT key genes in PC3 and SKOV3 cells incubated with CM CTRL or CM SEN for 72 h. ( E ) Invasion assay: PC3 and SKOV3 cells were incubated for 72 h with boiled and non-boiled CM CTRL or CM SEN and then allowed to invade for 16 h. Representative images of the filters are shown below the bar graphs (magnification 20×, scale bar 100 μm). ( F ) GC–MS analysis of metabolites in CM-CTRL or CM-SEN. Data reported are normalized to CM CTRL. * symbol indicates metabolite levels significantly increased both in senescent CM from HPFs and HOFs. Data are means ± SEM of three independent experiments. Statistical significance was assessed by unpaired Student t -test ( A , C , D , F ) or one-way Anova followed by Tukey’s multiple comparisons ( E ). * p < 0.05; ** p < 0.01; *** p < 0.001; ( ϕϕϕ p < 0.001; ϕϕϕϕ p < 0.0001; $$ p < 0.01; $$$ p < 0.001). In panel ( E ), * indicates CM SEN boiled vs. SEN non-boiled; ϕ indicates CM CTRL boiled vs. CM CTRL non-boiled; $ indicates CM SEN vs. CM CTRL.

Journal: Cells

Article Title: Senescent Stroma-Derived Glutamine: A Driver of Aggressiveness in Prostate and Ovarian Cancer Cells

doi: 10.3390/cells15090770

Figure Lengend Snippet: The metabolic component secreted by senescent HPFs and HOFs is involved in the acquisition of a pro-invasive phenotype in prostate and ovarian cancer cells. ( A ) Quantification of SA-β-Gal staining of human prostate fibroblasts (HPFs) and human ovarian fibroblasts (HOFs) treated for 24 h with 5 nM Docetaxel (DTX) and 20 μM cisplatin (CPT), respectively, and then cultured in drug-free medium for an additional 6 days. For each condition, images were taken from five randomly selected fields, and both the total number of cells and the number of blue (SA-β-Gal positive) cells were counted. Bar graphs represent the average ratio of positive cells to the total cell count. Representative images of the stained cells are shown below the bar graphs (magnification 20×, scale bar 100 μm). ( B ) Representative immunoblots of p16 and p21 protein levels in DTX-treated HPFs and CPT-treated HOFs. β-actin was used as loading control. ( C ) Invasion assay: PC3 and SKOV3 cells were incubated with CM CTRL and CM SEN conditioned media (CM) from senescent and non-senescent (CTRL) fibroblasts for 72 h and then seeded in Boyden chambers. Cells were allowed to invade for 16 h. Representative images of the filters are shown below the bar graphs (magnification 20×, scale bar 100 μm). ( D ) mRNA expression level of EMT key genes in PC3 and SKOV3 cells incubated with CM CTRL or CM SEN for 72 h. ( E ) Invasion assay: PC3 and SKOV3 cells were incubated for 72 h with boiled and non-boiled CM CTRL or CM SEN and then allowed to invade for 16 h. Representative images of the filters are shown below the bar graphs (magnification 20×, scale bar 100 μm). ( F ) GC–MS analysis of metabolites in CM-CTRL or CM-SEN. Data reported are normalized to CM CTRL. * symbol indicates metabolite levels significantly increased both in senescent CM from HPFs and HOFs. Data are means ± SEM of three independent experiments. Statistical significance was assessed by unpaired Student t -test ( A , C , D , F ) or one-way Anova followed by Tukey’s multiple comparisons ( E ). * p < 0.05; ** p < 0.01; *** p < 0.001; ( ϕϕϕ p < 0.001; ϕϕϕϕ p < 0.0001; $$ p < 0.01; $$$ p < 0.001). In panel ( E ), * indicates CM SEN boiled vs. SEN non-boiled; ϕ indicates CM CTRL boiled vs. CM CTRL non-boiled; $ indicates CM SEN vs. CM CTRL.

Article Snippet: Human prostate (PC3) and ovarian (SKOV3) cancer cell lines were obtained from ATCC (PC3: CVCL_E2RM; SKOV3: CVCL_0532).

Techniques: Staining, Cell Culture, Cell Characterization, Western Blot, Control, Invasion Assay, Incubation, Expressing, Gas Chromatography-Mass Spectrometry

Gln availability drives invasive ability in PC3 and SKOV3 cells. ( A ) Invasion assay: PC3 and SKOV3 cells were incubated for 72 h with DMEM in presence or absence of 2 mM Glutamine (Gln) and then seeded in Boyden chambers. Representative images of the filters are shown below the bar graphs (magnification 20×, scale bar 100 μm). ( B ) Invasion assay: PC3 and SKOV3 were incubated with CM CTRL and CM SEN for 72 h and with ASNase 1 U/mL during the last 48 h. Then, cells were seeded in Boyden chambers; ( C ) mRNA expression level of Gln transporter SLC1A5 in PC3 and SKOV3 cells incubated with CM CTRL or CM SEN for 72 h. ( D ) PC3 and SKOV3 cells were incubated for 72 h with CM CTRL or CM SEN. Then, GC–MS analysis was performed to measure Glu and Gln intracellular content. ( E ) Invasion assay: PC3 and SKOV3 cells were incubated for 72 h with DMEM and then seeded in a Boyden chamber in presence or absence of BPTES 1 μM for 16 h. Representative images of the filters are shown below the bar graphs (magnification 20×, scale bar 100 μm). ( F ) Invasion assay: PC3 and SKOV3 cells were incubated for 72 h with boiled and non-boiled CM; BPTES 1 μM was added during the final 16 h of incubation. Data are means ± SEM of three independent experiments. Statistical significance was assessed by Student t -test ( A , C – E ) or one-way Anova followed by Tukey’s multiple comparisons ( B , F ). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.001; ϕ p < 0.05; ϕϕ p < 0.01; ϕϕϕ p < 0.001; $ p < 0.05; $$ p < 0.01; $$$ p < 0.001. In panel ( E ), * indicates CM SEN + ASNase vs. CM SEN; ϕ indicates CM CTRL + ASNase vs. CM CTRL; $ indicates CM SEN vs. CM CTRL. In panel ( F ), * indicates treated CM SEN vs. CM SEN, ϕ indicates treated CM CTRL vs. CM CTRL, $ indicates CM SEN vs. CM CTRL.

Journal: Cells

Article Title: Senescent Stroma-Derived Glutamine: A Driver of Aggressiveness in Prostate and Ovarian Cancer Cells

doi: 10.3390/cells15090770

Figure Lengend Snippet: Gln availability drives invasive ability in PC3 and SKOV3 cells. ( A ) Invasion assay: PC3 and SKOV3 cells were incubated for 72 h with DMEM in presence or absence of 2 mM Glutamine (Gln) and then seeded in Boyden chambers. Representative images of the filters are shown below the bar graphs (magnification 20×, scale bar 100 μm). ( B ) Invasion assay: PC3 and SKOV3 were incubated with CM CTRL and CM SEN for 72 h and with ASNase 1 U/mL during the last 48 h. Then, cells were seeded in Boyden chambers; ( C ) mRNA expression level of Gln transporter SLC1A5 in PC3 and SKOV3 cells incubated with CM CTRL or CM SEN for 72 h. ( D ) PC3 and SKOV3 cells were incubated for 72 h with CM CTRL or CM SEN. Then, GC–MS analysis was performed to measure Glu and Gln intracellular content. ( E ) Invasion assay: PC3 and SKOV3 cells were incubated for 72 h with DMEM and then seeded in a Boyden chamber in presence or absence of BPTES 1 μM for 16 h. Representative images of the filters are shown below the bar graphs (magnification 20×, scale bar 100 μm). ( F ) Invasion assay: PC3 and SKOV3 cells were incubated for 72 h with boiled and non-boiled CM; BPTES 1 μM was added during the final 16 h of incubation. Data are means ± SEM of three independent experiments. Statistical significance was assessed by Student t -test ( A , C – E ) or one-way Anova followed by Tukey’s multiple comparisons ( B , F ). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.001; ϕ p < 0.05; ϕϕ p < 0.01; ϕϕϕ p < 0.001; $ p < 0.05; $$ p < 0.01; $$$ p < 0.001. In panel ( E ), * indicates CM SEN + ASNase vs. CM SEN; ϕ indicates CM CTRL + ASNase vs. CM CTRL; $ indicates CM SEN vs. CM CTRL. In panel ( F ), * indicates treated CM SEN vs. CM SEN, ϕ indicates treated CM CTRL vs. CM CTRL, $ indicates CM SEN vs. CM CTRL.

Article Snippet: Human prostate (PC3) and ovarian (SKOV3) cancer cell lines were obtained from ATCC (PC3: CVCL_E2RM; SKOV3: CVCL_0532).

Techniques: Invasion Assay, Incubation, Expressing, Gas Chromatography-Mass Spectrometry

Gln confers aggressiveness to prostate and ovarian cancer cells. ( A , B ) PC3 and SKOV3 cells were conditioned for 72 h with CM from HPFs and HOFs; BPTES 1 μM was added during the final 16 h of incubation. Then, cells were detached and grown as spheroids for 7 days. Volumes of prostatic and ovarian spheroids were calculated as described in M and M. Representative images of spheroids are shown below the bar graphs (magnification 20×, scale bar 100 μm). ( C ) Prostate and ovarian cancer cells were incubated for 72 h with or without 2 mM Gln. Then, cells were detached and grown as spheroids for 7 days. Volumes of prostatic and ovarian spheroids were calculated as described in M and M. Representative images of spheroids are shown below the bar graphs (magnification 20×, scale bar 100 μm). Data are represented as mean ± SEM of three independent experiments. Statistical significance was assessed by t -test. **** p < 0.0001.

Journal: Cells

Article Title: Senescent Stroma-Derived Glutamine: A Driver of Aggressiveness in Prostate and Ovarian Cancer Cells

doi: 10.3390/cells15090770

Figure Lengend Snippet: Gln confers aggressiveness to prostate and ovarian cancer cells. ( A , B ) PC3 and SKOV3 cells were conditioned for 72 h with CM from HPFs and HOFs; BPTES 1 μM was added during the final 16 h of incubation. Then, cells were detached and grown as spheroids for 7 days. Volumes of prostatic and ovarian spheroids were calculated as described in M and M. Representative images of spheroids are shown below the bar graphs (magnification 20×, scale bar 100 μm). ( C ) Prostate and ovarian cancer cells were incubated for 72 h with or without 2 mM Gln. Then, cells were detached and grown as spheroids for 7 days. Volumes of prostatic and ovarian spheroids were calculated as described in M and M. Representative images of spheroids are shown below the bar graphs (magnification 20×, scale bar 100 μm). Data are represented as mean ± SEM of three independent experiments. Statistical significance was assessed by t -test. **** p < 0.0001.

Article Snippet: Human prostate (PC3) and ovarian (SKOV3) cancer cell lines were obtained from ATCC (PC3: CVCL_E2RM; SKOV3: CVCL_0532).

Techniques: Incubation

GS silencing in senescent fibroblasts affects EMT program in prostate and ovarian cancer cells. ( A ) Immunoblot of GS protein levels in senescent and CTRL fibroblasts. Vinculin was used as loading control. ( B ) GS protein levels in senescent HPFs and HOFs following 48 h gene silencing. HSP90 immunoblot was performed to ensure equal loading. ( C ) Invasion assay: PC3 and SKOV3 cells were incubated for 72 h with CM from GS silenced HPFs and HOFs and then let to invade for 16 h. Representative images of filters are shown below the bar graphs (magnification 20×, scale bar 100 μm). ( D ) mRNA expression level of EMT markers in PC3 and SKOV3 cells after 72 h incubation with CM from GS-silenced fibroblasts. Data are mean ± SEM of three independent experiments. Statistical significance was assessed by Student t -test. t -test * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Journal: Cells

Article Title: Senescent Stroma-Derived Glutamine: A Driver of Aggressiveness in Prostate and Ovarian Cancer Cells

doi: 10.3390/cells15090770

Figure Lengend Snippet: GS silencing in senescent fibroblasts affects EMT program in prostate and ovarian cancer cells. ( A ) Immunoblot of GS protein levels in senescent and CTRL fibroblasts. Vinculin was used as loading control. ( B ) GS protein levels in senescent HPFs and HOFs following 48 h gene silencing. HSP90 immunoblot was performed to ensure equal loading. ( C ) Invasion assay: PC3 and SKOV3 cells were incubated for 72 h with CM from GS silenced HPFs and HOFs and then let to invade for 16 h. Representative images of filters are shown below the bar graphs (magnification 20×, scale bar 100 μm). ( D ) mRNA expression level of EMT markers in PC3 and SKOV3 cells after 72 h incubation with CM from GS-silenced fibroblasts. Data are mean ± SEM of three independent experiments. Statistical significance was assessed by Student t -test. t -test * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: Human prostate (PC3) and ovarian (SKOV3) cancer cell lines were obtained from ATCC (PC3: CVCL_E2RM; SKOV3: CVCL_0532).

Techniques: Western Blot, Control, Invasion Assay, Incubation, Expressing

Senescent stroma-derived Gln is uploaded by PC3. CTR and senescent HPFs were incubated for 24 h with 13 C glucose, then cell lysates and CM were collected for GC–MS analysis. PC3 cells were incubated with CM for further 24 h and the amount of 13 C Gln inside cells was measured. Data are represented as mean ± SEM of three independent experiments. Statistical significance was assessed by Student t -test. * p < 0.05.

Journal: Cells

Article Title: Senescent Stroma-Derived Glutamine: A Driver of Aggressiveness in Prostate and Ovarian Cancer Cells

doi: 10.3390/cells15090770

Figure Lengend Snippet: Senescent stroma-derived Gln is uploaded by PC3. CTR and senescent HPFs were incubated for 24 h with 13 C glucose, then cell lysates and CM were collected for GC–MS analysis. PC3 cells were incubated with CM for further 24 h and the amount of 13 C Gln inside cells was measured. Data are represented as mean ± SEM of three independent experiments. Statistical significance was assessed by Student t -test. * p < 0.05.

Article Snippet: Human prostate (PC3) and ovarian (SKOV3) cancer cell lines were obtained from ATCC (PC3: CVCL_E2RM; SKOV3: CVCL_0532).

Techniques: Derivative Assay, Incubation, Gas Chromatography-Mass Spectrometry

Gln-dependent NRF2/ETS1 pathway drives the invasive abilities of cancer cells. ( A ) Total ROS production was measured with a DCFDA probe in PC3 and SKOV3 cells after 72 h incubation with CM CTRL and CM SEN. ( B ) Total ROS production measured with DCFDA probe in PC3 and SKOV3 cells after incubation with boiled and non-boiled CM CTRL for 72 h in the presence or absence of ASNase 1 U/mL during the last 48 h. ( C ) Total ROS measured as described above. Cells were conditioned for 72 h, BPTES 1 μM was added during the final 16 h of incubation. ( D ) NRF2 protein level in PC3 and SKOV3 cells incubated either with DMEM in presence or absence of 2 mM Gln with CM CTRL and CM SEN. HSP90 immunoblot was performed to ensure equal loading. ( E ) ETS1 protein level in PC3 and SKOV3 cells incubated either with DMEM in presence or absence of 2 mM Gln or with CM CTRL or CM SEN. Actin was used as loading control. ( F ) GSH/GSSG ratio in prostate and ovarian cancer cells incubated for 72 h with CM CTRL or CM SEN. Data are normalized to protein content. ( G ) PC3 and SKOV3 cells were incubated for 72 h with CM CTRL or CM SEN; BPTES 1 μM was added during the final 16 h of incubation. Representative confocal microscopy images show ETS1 nuclear translocation. Bar graphs show the nuclear fluorescence intensity of ETS1 signal. Fluorescence intensity was quantified with ImageJ software (red: ETS1, blue: DAPI; scale bar 44 μm). Data are mean ± SEM of three independent experiments. Statistical significance was assessed by Student t -test ( A , C , F ) or one-way Anova followed by Tukey’s multiple comparisons ( B , G ). * p < 0.05; ** p < 0.01; φ p < 0.05; $$ p < 0.01. In panel ( G ), * indicates CM CTRL + BPTES vs. CM CTRL, $ indicates CM SEN vs. CM CTRL, φ indicates CM SEN + BPTES vs. CM SEN.

Journal: Cells

Article Title: Senescent Stroma-Derived Glutamine: A Driver of Aggressiveness in Prostate and Ovarian Cancer Cells

doi: 10.3390/cells15090770

Figure Lengend Snippet: Gln-dependent NRF2/ETS1 pathway drives the invasive abilities of cancer cells. ( A ) Total ROS production was measured with a DCFDA probe in PC3 and SKOV3 cells after 72 h incubation with CM CTRL and CM SEN. ( B ) Total ROS production measured with DCFDA probe in PC3 and SKOV3 cells after incubation with boiled and non-boiled CM CTRL for 72 h in the presence or absence of ASNase 1 U/mL during the last 48 h. ( C ) Total ROS measured as described above. Cells were conditioned for 72 h, BPTES 1 μM was added during the final 16 h of incubation. ( D ) NRF2 protein level in PC3 and SKOV3 cells incubated either with DMEM in presence or absence of 2 mM Gln with CM CTRL and CM SEN. HSP90 immunoblot was performed to ensure equal loading. ( E ) ETS1 protein level in PC3 and SKOV3 cells incubated either with DMEM in presence or absence of 2 mM Gln or with CM CTRL or CM SEN. Actin was used as loading control. ( F ) GSH/GSSG ratio in prostate and ovarian cancer cells incubated for 72 h with CM CTRL or CM SEN. Data are normalized to protein content. ( G ) PC3 and SKOV3 cells were incubated for 72 h with CM CTRL or CM SEN; BPTES 1 μM was added during the final 16 h of incubation. Representative confocal microscopy images show ETS1 nuclear translocation. Bar graphs show the nuclear fluorescence intensity of ETS1 signal. Fluorescence intensity was quantified with ImageJ software (red: ETS1, blue: DAPI; scale bar 44 μm). Data are mean ± SEM of three independent experiments. Statistical significance was assessed by Student t -test ( A , C , F ) or one-way Anova followed by Tukey’s multiple comparisons ( B , G ). * p < 0.05; ** p < 0.01; φ p < 0.05; $$ p < 0.01. In panel ( G ), * indicates CM CTRL + BPTES vs. CM CTRL, $ indicates CM SEN vs. CM CTRL, φ indicates CM SEN + BPTES vs. CM SEN.

Article Snippet: Human prostate (PC3) and ovarian (SKOV3) cancer cell lines were obtained from ATCC (PC3: CVCL_E2RM; SKOV3: CVCL_0532).

Techniques: Incubation, Western Blot, Control, Confocal Microscopy, Translocation Assay, Fluorescence, Software

The PDPK1/AKT/FLT DPI and tenovin-6 (T6) show high anti-cancer efficacy in murine tumoroids and human PCa cell lines (A) Dose-response curves for DPI (top) and T6 (bottom) for in vivo and in vitro Pten KO (left), Pten/Stat3 KO (middle), and Pten/Tp53 KO (right) tumoroids. Points represent means of technical duplicates per tumoroid line ( N = 3). Curve fitting was performed using GraphPad Prism 8.0.2. (B) Bar graphs showing means and ±SD of half-maximal inhibitory concentration (IC50) for DPI (top) and T6 (bottom) for in vivo and in vitro tumoroid lines of all genotypes ( N = 3). Statistical analysis was performed using GraphPad Prism 8.0.2 (one-way ANOVA, Tukey’s test). p > 0.05 if not specified otherwise, ∗ p ≤ 0.05. (C) Bar graphs depicting means and ±SD of IC50 values of DPI (left) and T6 (right) on human PCa cell lines. 22RV1: primary PCa; LNCaP: metastatic PCa; DU145, PC3: metastatic castration-resistant PCa ( N = 3). Statistical analysis was performed using GraphPad Prism 8.0.2 (one-way ANOVA). p > 0.05 if not specified otherwise, ∗ p ≤ 0.05; ∗∗ p ≤ 0.01. (D) Heatmaps of synergy scores calculated with the highest single agent (HSA) model for DPI and enzalutamide (left), and T6 and enzalutamide (right) on the human LNCaP cell line. Values > 0 represent synergistic effects, and values < 0 represent antagonistic effects. IC50 concentrations of respective compounds are underlined ( N = 3). See also .

Journal: Cell Reports Methods

Article Title: Biobank of genetically defined murine prostate cancer tumoroids uncovers oncogenic pathways and drug vulnerabilities driven by PTEN-loss

doi: 10.1016/j.crmeth.2026.101370

Figure Lengend Snippet: The PDPK1/AKT/FLT DPI and tenovin-6 (T6) show high anti-cancer efficacy in murine tumoroids and human PCa cell lines (A) Dose-response curves for DPI (top) and T6 (bottom) for in vivo and in vitro Pten KO (left), Pten/Stat3 KO (middle), and Pten/Tp53 KO (right) tumoroids. Points represent means of technical duplicates per tumoroid line ( N = 3). Curve fitting was performed using GraphPad Prism 8.0.2. (B) Bar graphs showing means and ±SD of half-maximal inhibitory concentration (IC50) for DPI (top) and T6 (bottom) for in vivo and in vitro tumoroid lines of all genotypes ( N = 3). Statistical analysis was performed using GraphPad Prism 8.0.2 (one-way ANOVA, Tukey’s test). p > 0.05 if not specified otherwise, ∗ p ≤ 0.05. (C) Bar graphs depicting means and ±SD of IC50 values of DPI (left) and T6 (right) on human PCa cell lines. 22RV1: primary PCa; LNCaP: metastatic PCa; DU145, PC3: metastatic castration-resistant PCa ( N = 3). Statistical analysis was performed using GraphPad Prism 8.0.2 (one-way ANOVA). p > 0.05 if not specified otherwise, ∗ p ≤ 0.05; ∗∗ p ≤ 0.01. (D) Heatmaps of synergy scores calculated with the highest single agent (HSA) model for DPI and enzalutamide (left), and T6 and enzalutamide (right) on the human LNCaP cell line. Values > 0 represent synergistic effects, and values < 0 represent antagonistic effects. IC50 concentrations of respective compounds are underlined ( N = 3). See also .

Article Snippet: Human PC3 metastatic PCa cell line , ATCC , CRL-1435; RRID:CVCL_0035.

Techniques: In Vivo, In Vitro, Concentration Assay

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.

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

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.

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.

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

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.

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.

Techniques: Marker, Expressing, Staining

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.

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.

Techniques: In Vivo, Staining, Immunohistochemical staining, Immunostaining

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.

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.

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

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.

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.

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

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