human prostate carcinoma cell line du145  (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