Journal: bioRxiv

Article Title: Improve the Efficacy of B7-H3-Targeting Antibody-Drug Conjugate DS-7300a in TP53-deficient Tumors by Inducing Ferroptosis

doi: 10.64898/2026.05.05.722958

Figure Lengend Snippet: A. Schematic of the mechanism of action of B7-H3-targeted antibody-drug conjugate, DS-7300a equipped with a DNA topoisomerase I (TOP1) inhibitor, Dxd. B-C. Western blot of p53 and dose-response curves of DS-7300a in TP53-WT versus TP53-KO LNCaP (B) and 22Rv1 (C) cells. D. TP53 was reintroduced into TP53-KO LNCaP cells, followed by Western blot and DS-7300a dose-response assays. E. IC 50 determination of DS-7300a in diverse PCa cell lines containing wildtype or mutated TP53 . Cells were treated with the indicated concentrations of DS-7300a for 6 days in all experiments with at least three replicates. Data represent the mean ± standard deviation of triplicates or otherwise stated. IC 50 values were calculated using GraphPad Prism version 10.4.1.

Article Snippet: 2×10 6 LNCaP cells with or without TP53 knockouts [ ] were subcutaneously injected into both flanks of seven-week-old male SCID mice (CB17 Charles River).

Techniques: Western Blot, Standard Deviation

Journal: bioRxiv

Article Title: Improve the Efficacy of B7-H3-Targeting Antibody-Drug Conjugate DS-7300a in TP53-deficient Tumors by Inducing Ferroptosis

doi: 10.64898/2026.05.05.722958

Figure Lengend Snippet: A. Schematic of experimental design using xenograft models. 2×10 6 TP53-WT and TP53-KO LNCaP cells were subcutaneously injected into both flanks of seven-week-old male SCID mice. When tumors reached approximately 75mm 3 , tumor-bearing mice were randomized for treatment of ABS vehicle or DS-7300a (0.5 mg/kg; i.v., biweekly, twice). B-C. Tumor growth over time (B) and tumor weights at the endpoint (C) in xenograft models are shown. D. Representative images and quantification of Ki67 IHC staining in xenograft tumors after treatments. Scale bar = 50um. E. Schematic of experimental design using humanized B7-H3 models. Human B7-H3 gene was introduced into the syngeneic DX1 PCa cell line to generate DX1-hCD276 cells. 2×10 6 DX1-hCD276 cells were subcutaneously injected into both flanks of male B-hB7-H3 mice, followed by treatment of ABS vehicle control or DS-7300a (0.5 mg/kg; i.v., biweekly, twice). F. B7-H3 protein expression of DX1-hCD276 cells was verified by Flow Cytometry. G-H. Tumor growth over time (G) and tumor weights at the endpoint (H) of humanized B7-H3 PCa models after treatment. Data represent the mean ± standard deviation of triplicates or otherwise stated. Statistics were calculated using GraphPad Prism version 10.4.1. P values determined by unpaired two-tailed t test (B, C, H) or by two-way ANOVA analysis (H); *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; ns, not significant.

Article Snippet: 2×10 6 LNCaP cells with or without TP53 knockouts [ ] were subcutaneously injected into both flanks of seven-week-old male SCID mice (CB17 Charles River).

Techniques: Injection, Immunohistochemistry, Control, Expressing, Flow Cytometry, Standard Deviation, Two Tailed Test

Journal: bioRxiv

Article Title: Improve the Efficacy of B7-H3-Targeting Antibody-Drug Conjugate DS-7300a in TP53-deficient Tumors by Inducing Ferroptosis

doi: 10.64898/2026.05.05.722958

Figure Lengend Snippet: A. Dose-response curves and IC 50 of DXd in LNCaP with or without TP53 KO. B. Re-introducing TP53 ORF sensitizes TP53-KO LNCaP cells to DXd. C. Comparison of DXd IC 50 in various PCa cell lines containing wildtype or mutated TP53. D. Western blot determining the expression of p53, p21, and DNA damage response signaling in TP53-WT and TP53-KO cells treated with DXd. E-G. mRNA expression levels of TP53 (E) and its direct target genes involved in senescence (F) and apoptosis (G) in TP53-WT and TP53-KO cells after DXd treatment, determined by qPCR. H-I. TP53-WT and TP53-KO LNCaP cells were treated with DXd at indicated concentrations, followed by Annexin V and DAPI analysis using Flow (H) or beta-galactosidase staining (I). Scale bar = 50um. Cells were treated with DXd for 48 hours in all experiments. Data represent the mean ± standard deviation of triplicates. IC 50 values were calculated using GraphPad Prism version 10.4.1. P values determined by one-way ANOVA analysis; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; ns, not significant.

Article Snippet: 2×10 6 LNCaP cells with or without TP53 knockouts [ ] were subcutaneously injected into both flanks of seven-week-old male SCID mice (CB17 Charles River).

Techniques: Comparison, Western Blot, Expressing, Staining, Standard Deviation

Journal: bioRxiv

Article Title: Improve the Efficacy of B7-H3-Targeting Antibody-Drug Conjugate DS-7300a in TP53-deficient Tumors by Inducing Ferroptosis

doi: 10.64898/2026.05.05.722958

Figure Lengend Snippet: A. Western blot determining the expression of p53, p21, and DNA damage response signaling in TP53-WT and TP53-KO cells treated with DS-7300a for 96h. B-D. mRNA expression levels of TP53 (B) and its direct target genes involved in senescence (C) and apoptosis (D) in TP53-WT and TP53-KO cells after DS-7300a treatment for 96h, determined by qPCR. E-F. TP53-WT and TP53-KO cells were treated with DS-7300a for 72h, followed by apoptosis analysis (E) or beta-galactosidase staining (F). Scale bar = 50um. G . Representative images and quantification of IHC staining of indicated markers in TP53-WT and TP53-KO LNCaP xenograft tumors treated with 0.5 mg/kg DS-7300a. Scale bar = 50um. Data represent the mean ± standard deviation of triplicates. Statistics were calculated using GraphPad Prism version 10.4.1. P values determined by one-way ANOVA analysis; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; ns, not significant.

Article Snippet: 2×10 6 LNCaP cells with or without TP53 knockouts [ ] were subcutaneously injected into both flanks of seven-week-old male SCID mice (CB17 Charles River).

Techniques: Western Blot, Expressing, Staining, Immunohistochemistry, Standard Deviation

Journal: bioRxiv

Article Title: Improve the Efficacy of B7-H3-Targeting Antibody-Drug Conjugate DS-7300a in TP53-deficient Tumors by Inducing Ferroptosis

doi: 10.64898/2026.05.05.722958

Figure Lengend Snippet: A-B. GPX4 expression in TP53-KO LNCaP cells treated with DS-7300a in vitro (A) and TP53-KO LNCaP xenograft tumors after DS-7300a treatment in vivo (B), determined by Western blot and qPCR. C. Lipid peroxidation levels in TP53-KO LNCaP cells treated with DS-7300a or/and RSL-3 with or without ferroptosis inhibitor ferrostatin. D-G. Dose-response matrix (left) and drug interaction plots (right) revealing the synergistic effects of DS-7300a and RSL-3 in various TP53- deficient PCa cell lines: TP53-KO LNCaP (D), TP53-KO 22Rv1 (E), DU145 (TP53-mutated, F), and LAPC4 (TP53-mutated, G). Drug interaction landscapes and synergy scores were generated using SynergyFinder. H. Cell viability assay of TP53-KO LNCaP cells treated with DS-7300a or/and RSL-3 with or without ferroptosis inhibitor ferrostatin. Data represent the mean ± standard deviation of quadruplicates. Data represent the mean ± standard deviation of triplicates. Statistics were calculated using GraphPad Prism version 10.4.1. P values determined by unpaired two-tailed t test (B) or one-way ANOVA analysis (A, C, H). *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; ns, not significant.

Article Snippet: 2×10 6 LNCaP cells with or without TP53 knockouts [ ] were subcutaneously injected into both flanks of seven-week-old male SCID mice (CB17 Charles River).

Techniques: Expressing, In Vitro, In Vivo, Western Blot, Generated, Viability Assay, Standard Deviation, Two Tailed Test

Journal: bioRxiv

Article Title: Improve the Efficacy of B7-H3-Targeting Antibody-Drug Conjugate DS-7300a in TP53-deficient Tumors by Inducing Ferroptosis

doi: 10.64898/2026.05.05.722958

Figure Lengend Snippet: A. Schematic of experimental design of combination treatment in humanized B7-H3 models. 2×10 6 DX1-hCD276 cells were subcutaneously injected into both flanks of male B-hB7-H3 mice, followed by treatment of single agents or combination of DS-7300a (2 mg/kg; i.v.; once) or/and JKE-1674 (10 mg/kg; oral gavage, every two days). B-D. Tumor growth over time (B), tumor weights at the endpoint (C), and mice body weight changes (D) of humanized B7-H3 PCa models after treatments. E. Schematic of experimental design in xenograft models. 2×10 6 TP53-KO LNCaP cells were subcutaneously injected into both flanks of SCID mice. When tumors reached approximately 75mm 3 , tumor-bearing mice were randomized for combination treatment (3 mg/kg DS-7300a, i.v., biweekly, twice; 15 mg/kg JKE-1674, oral gavage, every two days) or ABS vehicle control. F-G. Tumor growth over time (F) and H&E staining at the endpoint (G) in xenograft models are shown. Scale bar = 2 mm. H. Representative images and quantification of 4-HNE IHC staining in xenograft tumors after treatments. Scale bar = 50um I. Schematic working model. Data represent the mean ± standard deviation of triplicates or otherwise stated. P values determined by unpaired two-tailed t test (F, H) or one-way ANOVA analysis (B, C, D). *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; ns, not significant.

Article Snippet: 2×10 6 LNCaP cells with or without TP53 knockouts [ ] were subcutaneously injected into both flanks of seven-week-old male SCID mice (CB17 Charles River).

Techniques: Injection, Control, Staining, Immunohistochemistry, Standard Deviation, Two Tailed Test

Journal: Oncology Reports

Article Title: Sphingosine-1-phosphate receptor 1 enhances olfactory receptor 51E1-mediated inhibition of proliferation via Src/JNK signaling in prostate cancer cells

doi: 10.3892/or.2026.9103

Figure Lengend Snippet: OR51E1 agonists reduce LNCaP cell viability, but expression alone does not predict patient outcome. (A) LNCaP cells were treated with increasing concentrations (0.1–1 mM) of NA or BA for 48 h, and cell viability was measured using the Cell Counting Kit-8 assay. Values were normalized to untreated controls. (B) The effect of NA on cell viability was assessed in control and OR51E1 knockout LNCaP cells after 48 h of NA (0.5 mM) treatment. (C) Relative OR51E1 mRNA expression in LNCaP, DU145, and PC3 prostate cancer cell lines was assessed by reverse transcription-quantitative PCR. Expression levels were normalized to β-actin. Representative PCR products were visualized by agarose gel electrophoresis. (D) LNCaP, DU145 and PC3 cells were cultured with various concentrations of NA for 48 h, and cell viability was measured. Data represent the mean ± SEM of three independent experiments. Statistical significance was determined using an unpaired Student's t-test. (E) OR51E1 expression across pathological stages of prostate cancer. OR51E1 mRNA expression levels were compared between normal prostate tissues from the GTEx dataset and prostate adenocarcinoma samples from TCGA stratified by pathological stage: Stage I (T2b), Stage II (T2b and T2c), Stage III (T3a and 3b), and Stage IV (T4). Transcript expression values (RSEM TPM) were obtained from the UCSC Xena Browser using the TCGA-TARGET-GTEx TOIL RNA-seq recompute dataset. Statistical significance was determined using an unpaired Student's t-test. (F and G) Kaplan-Meier survival curves for (F) overall survival and (G) progression-free interval stratified by OR51E1 expression levels. Red and blue lines indicate high- and low-expression groups, respectively. Survival probabilities were compared using the log-rank test, and P-values are shown in each panel. *P<0.05, **P<0.01, ***P<0.001 and ****P<0.0001. OR51E1, olfactory receptor 51E1; BA, butyric acid; NA, nonanoic acid; GTEx, Genotype-Tissue Expression; TCGA, The Cancer Genome Atlas; ns, not significant.

Article Snippet: LNCaP cells were obtained from the Korean Cell Line Bank, and Du145 and PC3 cells were purchased from the American Type Culture Collection.

Techniques: Expressing, Cell Counting, Control, Knock-Out, Reverse Transcription, Real-time Polymerase Chain Reaction, Agarose Gel Electrophoresis, Cell Culture, RNA Sequencing, Olfactory

Journal: Oncology Reports

Article Title: Sphingosine-1-phosphate receptor 1 enhances olfactory receptor 51E1-mediated inhibition of proliferation via Src/JNK signaling in prostate cancer cells

doi: 10.3892/or.2026.9103

Figure Lengend Snippet: S1PR1 enhances OR51E1-mediated cytotoxicity and apoptosis in LNCaP cells. (A) LNCaP cells were transfected with an empty vector, S1PR1, or DRD2. After 48 h, cells were treated with NA (0.5 mM) for an additional 48 h, and cell viability was measured using the Cell Counting Kit-8 assay. (B) LNCaP cells were transduced with lentivirus encoding S1PR1 and selected with puromycin (1 µg/ml). Cell viability of parental LNCaP and LNCaP-S1PR1 cells was measured 48 h after treatment with increasing concentrations of NA (0.1–1 mM). (C) Apoptosis was assessed in LNCaP cells treated with NA for 48 h using annexin V/PI staining. Apoptotic cells were quantified as the percentage of annexin V-positive cells (early apoptosis) and annexin V/PI double-positive cells (late apoptosis) relative to the total cell population. (D) Caspase-3 activation was measured by flow cytometry following NA treatment. (E) PARP1 cleavage was analyzed by western blotting in NA-treated cells. Band intensities were quantified using ImageJ software. Data represent the mean ± SEM of at least three independent experiments. Statistical significance was determined using an unpaired Student's t-test. *P<0.05, **P<0.01 and ***P<0.001. S1PR1, sphingosine-1-phosphate receptor 1; OR51E1, olfactory receptor 51E1; DRD2, dopamine receptor D2; NA, nonanoic acid; PI, propidium iodide; ns, not significant.

Article Snippet: LNCaP cells were obtained from the Korean Cell Line Bank, and Du145 and PC3 cells were purchased from the American Type Culture Collection.

Techniques: Transfection, Plasmid Preparation, Cell Counting, Transduction, Staining, Activation Assay, Flow Cytometry, Western Blot, Software, Olfactory

Journal: Oncology Reports

Article Title: Sphingosine-1-phosphate receptor 1 enhances olfactory receptor 51E1-mediated inhibition of proliferation via Src/JNK signaling in prostate cancer cells

doi: 10.3892/or.2026.9103

Figure Lengend Snippet: Activation of S1PR1 enhances OR51E1-mediated apoptosis in LNCaP cells. (A) LNCaP and LNCaP-S1PR1 cells were treated with NA, TC-G1006 (a selective S1PR1 agonist), or both for 48 h, and apoptosis was assessed using annexin V/PI staining. (B) LNCaP and LNCaP-S1PR1 cells were preincubated with DMSO, PTX, or NIBR-0213 (a selective S1PR1 antagonist) for 1 h and then treated with NA for 48 h. Cell viability was measured using the CCK-8 assay. Data represent the mean ± SEM of at least three independent experiments. Statistical analysis was performed using two-way ANOVA with Tukey's multiple comparison test. *P<0.05, **P<0.01 and ****P<0.0001. S1PR1, sphingosine-1-phosphate receptor 1; OR51E1, olfactory receptor 51E1; NA, nonanoic acid; PI, propidium iodide; DMSO, dimethyl sulfoxide; PTX, pertussis toxin; ns, not significant.

Article Snippet: LNCaP cells were obtained from the Korean Cell Line Bank, and Du145 and PC3 cells were purchased from the American Type Culture Collection.

Techniques: Activation Assay, Staining, CCK-8 Assay, Comparison, Olfactory

Journal: Oncology Reports

Article Title: Sphingosine-1-phosphate receptor 1 enhances olfactory receptor 51E1-mediated inhibition of proliferation via Src/JNK signaling in prostate cancer cells

doi: 10.3892/or.2026.9103

Figure Lengend Snippet: Co-expression of S1PR1 amplifies OR51E1-Src-JNK signaling and suppression of proliferation. (A and B) Inhibitor profiling reveals signaling pathways involved in NA-induced reduction of LNCaP cell viability. LNCaP cells were preincubated for 1 h with (A) canonical OR pathway inhibitors (SQ-22536, 100 µM; H-89, 3 µM; ESI-09, 3 µM; SU6656, 10 µM) or with (B) other, less well-characterized pathway inhibitors (SU6656, 10 µM; Dasatinib, 10 nM; SP600125, 3 µM; SB203580, 3 µM; U0126, 10 µM; AG-490, 10 µM), followed by treatment with NA for 48 h. SQ-22536 and SU6656 were included in both panels for comparison. Cell viability was measured using the CCK-8 assay. Data represent the mean ± SEM of at least three independent experiments. Statistical analysis was performed using an unpaired Student's t-test. Hash symbols (#) indicate statistical significance between vehicle- and NA-treated cells, whereas asterisks (*) indicate statistical significance between cells treated with NA alone and those co-treated with NA and the indicated inhibitors. (C) Western blot analysis of JNK activation in LNCaP and LNCaP-S1PR1 cells treated with NA for 1 h. Protein levels of p-JNK and total JNK were analyzed. (D) Effect of Src inhibition on NA-induced JNK activation. LNCaP cells were pretreated with the Src inhibitor SU6656 (10 µM) for 1 h and then treated with NA for 1 h. Protein levels of p-JNK and total JNK were assessed. Band intensities were quantified using ImageJ software. Data represent the mean ± SEM of three independent experiments. Statistical significance was determined using an unpaired Student's t-test. (E) Effect of Src inhibition on NA-mediated suppression of proliferation. LNCaP and LNCaP-S1PR1 cells were pretreated with SU6656 for 1 h, and cell viability was measured using the CCK-8 assay after NA treatment. Data represent the mean ± SEM of at least three independent experiments and were analyzed using two-way ANOVA with Tukey's multiple comparison test. (F) Effect of Src inhibition on NA-induced apoptosis. LNCaP and LNCaP-S1PR1 cells were pretreated with dasatinib (100 nM) for 1 h and then treated with NA. Apoptosis was assessed using annexin V/PI staining. Data represent the mean ± SEM of three independent experiments and were analyzed using two-way ANOVA with Tukey's multiple comparison test. ### P<0.001 and #### P<0.0001; *P<0.05, **P<0.01, ***P<0.001 and ****P<0.0001. S1PR1, sphingosine-1-phosphate receptor 1; OR51E1, olfactory receptor 51E1; NA, nonanoic acid; CCK-8, Cell Counting Kit-8; p-, phosphorylated; PI, propidium iodide; ns, not significant.

Article Snippet: LNCaP cells were obtained from the Korean Cell Line Bank, and Du145 and PC3 cells were purchased from the American Type Culture Collection.

Techniques: Expressing, Protein-Protein interactions, Comparison, CCK-8 Assay, Western Blot, Activation Assay, Inhibition, Software, Staining, Olfactory, Cell Counting

Journal: PLOS Computational Biology

Article Title: CASER: A semi-supervised model with multi-omics data integration prioritizes cancer-associated epigenetic regulator genes

doi: 10.1371/journal.pcbi.1014253

Figure Lengend Snippet: The robust rank aggregation (RRA) scores of negative selections are shown for different gene sets in (A) LNCaP and (B) MDA-MB-231 cells. The Chronos dependency scores are shown for different gene sets in (C) LNCaP and (D) TNBC cancer cell. The RRA scores of negative selection were calculated using MAGeCK. The higher -log 10 (RRA) values indicate a greater effect on cell survival after gene knockdown. The lower DEMETER and Chronos score indicate a greater effect on cell survival after gene knockdown. The genes in this analysis are both cancer genes and ERGs. Non-cancer ERs refer to the ER genes that are also neutral genes. P -value is calculated by Wilcoxon rank-sum one-tailed test.

Article Snippet: The human melanoma cell line SK-mel-2, clear cell renal cell carcinoma cell line Caki-1, breast cancer cell line MDA-MB-231, and prostate cancer cell line LNCaP, along with their corresponding complete median, were purchased from Procell Life Science & Technology Co., Ltd. (Wuhan, China).

Techniques: Selection, Knockdown, One-tailed Test

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 LNCaP metastatic PCa cell line , ATCC , CRL-1740; RRID:CVCL_A4BQ.

Techniques: In Vivo, In Vitro, Concentration Assay