human prostate cancer cell lines  (ATCC)

Journal: bioRxiv

Article Title: Co-Targeting Nuclear Export and Translation Initiation Uncovers a Therapeutic Vulnerability in Lethal Prostate Cancer

doi: 10.64898/2026.05.04.722693

Figure Lengend Snippet: (A) Schematic of the quantitative high-throughput screening (qHTS) workflow using the Mechanism Interrogation PlatE (MIPE) small-molecule library to measure drug activity and synergy. Created with Biorender.com. (B) Heatmap of Z-normalized area under the curve (Z-AUC) values derived from 11-point dose-response viability measurements (CellTiter-Glo) in AR-V7-expressing LNCaP-95 (48 h readout) and VCaP-CR (72 h readout) cells, with the Pearson correlation coefficient between cell types indicated. (C) Mean Z-AUC values across LNCaP-95 and VCaP-CR cells for each compound, ranked in descending order and annotated by primary molecular target (“i” denotes inhibitor). (D) Drug Target Set Enrichment Analysis (DSEA) demonstrating significant enrichment of XPO1-targeting compounds among the most active agents when compounds are ranked by Z-AUC. (E) Heatmap of Z-AUC values for XPO1 inhibitors (Eltanexor, Selinexor, Leptomycin B, and KPT-276) across the indicated panel of prostate cancer cells (LNCaP, VCaP, 22Rv1, LNCaP-Abl, LNCaP-95, VCaP-CR, DU145, and PC3). (F) Single agent dose-response curves displaying percent cell viability following Eltanexor treatment across an 11-point concentration range (0.8 nM-45 μM; 1:3 dilution series) in representative castration-sensitive, castration-resistant, and aggressive variant prostate cancer cells. (G) As in (F), dose-response curves for Selinexor. (H-K) Clinical prostate cancer samples were stratified into quartiles based on XPO1 expression, ranging from low (Q1) to high (Q4). An AR-V7 target gene score was calculated as the summed expression of genes from the Sharp et al. (2019) signature (n = 59). Differences across quartiles were assessed using the Kruskal-Wallis test, followed by Dunn’s post hoc testing with Benjamini-Hochberg correction. Analyses were performed in primary prostate cancer cohorts from (H) DKFZ and (I) TCGA, and metastatic castration-resistant prostate cancer cohorts from (J) SU2C/PCF and (K) UW/FH. Statistical significance is indicated (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001). (L) Kaplan-Meier overall survival analysis in the SU2C/PCF cohort comparing low (Q1) versus high (Q4) XPO1 expression. Hazard ratio (HR) per standard deviation (SD) with 95% confidence interval (CI) and Cox proportional hazards p-value are shown; numbers at risk are indicated below. See also Figure S1.

Article Snippet: The human prostate cancer cell lines 22Rv1 (RRID: CVCL_1045), DU145 (RRID: CVCL_0105), PC3 (RRID: CVCL_0035), VCaP (RRID: CVCL_2235), and LNCaP (RRID: CVCL_1379) were obtained from the American Type Culture Collection (ATCC).

Techniques: High Throughput Screening Assay, Activity Assay, Derivative Assay, Expressing, Concentration Assay, Variant Assay, Standard Deviation

Journal: bioRxiv

Article Title: Co-Targeting Nuclear Export and Translation Initiation Uncovers a Therapeutic Vulnerability in Lethal Prostate Cancer

doi: 10.64898/2026.05.04.722693

Figure Lengend Snippet: (A) Unsupervised hierarchical clustering of a 42-drug all-versus-all combination screen based on correlation of Excess Highest Single Agent (Excess HSA) scores, identifying clusters of compounds with similar combination profiles. Compounds selected based on single agent activity and mechanistic relevance were tested across 10 × 10 dose matrices (n = 861 combinations) in LNCaP-95 and VCaP-CR cells, with Excess HSA values averaged across both models. The black box highlights clustering of combination profiles involving the XPO1 inhibitors Eltanexor and Selinexor. (B) Excess HSA synergy scores overlaid onto the same unsupervised clustering heatmap, highlighting synergistic (red) and antagonistic (blue) interactions. The black boxes highlight strong synergy between XPO1 inhibitors (Eltanexor, Selinexor) and the EIF4A1 inhibitor Zotatifin. (C) Circos plot displaying the most synergistic drug combinations (Excess HSA < -2000) identified in the screen. (D) Top 10 most synergistic combinations with Eltanexor, ranked by mean Excess HSA across LNCaP-95 and VCaP-CR cells. (E) Top 10 most synergistic combinations with Zotatifin, ranked by mean Excess HSA across LNCaP-95 and VCaP-CR cells. (F) Targeted combination screening evaluating XPO1 and EIF4A1 inhibition across prostate cancer models. Eltanexor was combined with EIF4A1 inhibitors (Zotatifin, CR-1-31-B, Rocaglamide A, and C5-desmethyl PatA), and Zotatifin was combined with XPO1 inhibitors (Eltanexor, Selinexor, Verdinexor, and Leptomycin B) across a range of starting concentrations. Excess HSA scores were calculated to quantify synergy across DU145, PC3, LNCaP-95, LNCaP, 22Rv1, VCaP-CR, and VCaP cells. (G-H) Focused 10 × 10 dose-response matrices for the Eltanexor + Zotatifin combination extracted from the combination screen and analyzed using SynergyFinder in (G) LNCaP-95 and (H) VCaP-CR cells. Heatmaps display percent inhibition of cell viability, with corresponding three-dimensional surface plots depicting Highest Single Agent (HSA) synergy scores. (I-K) Validation of Eltanexor + Zotatifin synergy in patient-derived organoid models. Percent inhibition heatmaps and three-dimensional HSA synergy surface plots are shown for (I) LuCaP167-CR, (J) LuCaP136-CR, and (K) Lym1 organoids. Data represent the mean from n=3 replicates. See also Figure S2-4.

Article Snippet: The human prostate cancer cell lines 22Rv1 (RRID: CVCL_1045), DU145 (RRID: CVCL_0105), PC3 (RRID: CVCL_0035), VCaP (RRID: CVCL_2235), and LNCaP (RRID: CVCL_1379) were obtained from the American Type Culture Collection (ATCC).

Techniques: Activity Assay, Inhibition, Biomarker Discovery, Derivative Assay

Journal: bioRxiv

Article Title: Co-Targeting Nuclear Export and Translation Initiation Uncovers a Therapeutic Vulnerability in Lethal Prostate Cancer

doi: 10.64898/2026.05.04.722693

Figure Lengend Snippet: (A) Time-course Caspase-Glo screening in which Zotatifin was combined with XPO1 inhibitors (Eltanexor, Selinexor, Verdinexor, and Leptomycin B) in LNCaP-95 and VCaP-CR cells using 10 × 10 dose-response matrices. Caspase activity was measured every 2 hours from 2 to 24 hours post-treatment, and synergy was quantified as Excess Highest Single Agent (Excess HSA). (B-D) Focused analysis of the Zotatifin-Eltanexor combination at (B) 16, (C) 20, and (D) 24 hours in LNCaP-95 cells. Heatmaps display the percent apoptotic response measured by Caspase-Glo, with corresponding three-dimensional surface plots depicting Highest Single Agent (HSA) synergy scores. (E-F) Live-cell imaging (Incucyte) of LNCaP-95, VCaP-CR, and 22Rv1 cells treated with DMSO, single agents, or the Eltanexor + Zotatifin combination. Images were acquired every 4 hours over 7 days. (E) Cell proliferation quantified as percent confluence normalized to 0 h. (F) Apoptotic index quantified using Caspase-3/7 fluorescent dye, normalized to cell confluence. Data represent mean ± SEM from n = 2-3 replicates. (G) Representative Incucyte images of LNCaP-95 cells grown as 3D spheroids and treated with the Eltanexor + Zotatifin combination. Images show Caspase-3/7 fluorescence (green) with corresponding phase-contrast images at 0, 48, and 120 hours. Scale bar, 400 μm. (H) As in (G), representative Incucyte images of VCaP-CR 3D cell aggregates.

Article Snippet: The human prostate cancer cell lines 22Rv1 (RRID: CVCL_1045), DU145 (RRID: CVCL_0105), PC3 (RRID: CVCL_0035), VCaP (RRID: CVCL_2235), and LNCaP (RRID: CVCL_1379) were obtained from the American Type Culture Collection (ATCC).

Techniques: Activity Assay, Live Cell Imaging, Fluorescence

Journal: bioRxiv

Article Title: Salvianolic acids are natural senolytics and increase lifespan in old age

doi: 10.64898/2026.04.29.721790

Figure Lengend Snippet: ( a ) A schematic workflow illustrating the preclinical therapeutic procedure. Two weeks after subcutaneous inoculation and in vivo uptake of PC3/PSC27 recombinants, severe combined immunodeficient (M-NSG) mice were subject to either single agent or combinatorial treatment in a metronomic schedule of several cycles. The illustration was created in BioRender. ( b ) Comparative statistics of tumor end volumes. PC3 cancer cells were inoculated either alone or combined with PSC27 stromal cells before being implanted subcutaneously to the hind flank of M-NSG animals, which were administered with MIT and SAA, either alone or in combination. ( c ) Representative images of in vivo senescence in tumor foci by SA-β-gal staining. Scale bar, 50 μm. ( d ) Comparative statistics of tumor senescence as described in ( c ). ( e ) Transcriptional analysis of a subset of SASP factors expressed in epithelial vs. stromal cells acquired from tumor foci after laser capture microdissection (LCM) to isolate stromal and cancer cells, respectively. Signals were normalized to that of the sample with lowest value in the placebo group. ( f ) Transcriptional analysis of SASP factors and two canonical senescence biomarkers p16 INK4a and p21 CIP1 . ( g ) Statistical assessment of DDR and cellular apoptosis in tumors. Values are shown as the percentage of cells positively stained by IF or IHC specific to γH2AX or cleaved caspase 3 (CCL3), respectively. ( h ) Representative images of IHC staining for CCL3 at the completion of treatment regimens. Scale bar, 50 μm. ( i ) Comparative survival of animals sacrificed upon development of advanced bulky disease. Survival duration was calculated from tissue recombinant injection until death. P values were calculated by a two-sided log-rank (Mantel-Cox) test. DDR, DNA damage response. IF, immunofluorescence. IHC, immunohistochemistry. Data in b , d , e , f and g are shown as mean ± SD and representative of 3 independent biological replicates, with P values calculated by Student’s t -tests. ^, P > 0.05; *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

Article Snippet: The human prostate epithelial cancer cell line, PC3, was from ATCC and cultured with F-12K media (10% FBS).

Techniques: In Vivo, Staining, Laser Capture Microdissection, Immunohistochemistry, Recombinant, Injection, Immunofluorescence

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