Journal: Cellular & Molecular Biology Letters

Article Title: The marine-derived HIF-1α inhibitor, Yardenone 2, reduces prostate cancer cell proliferation by targeting HIF-1 target genes

doi: 10.1186/s11658-024-00617-2

Figure Lengend Snippet: Effect of Sodwanones and Yardenones on HIF-1α stabilization. A P69, DU145, PC3, and 786-O cells were subjected to hypoxia 1% O 2 (Hx 1%) for 24 and 48h. Cell lysates were analyzed by immunoblotting for HIF-1α. Tubulin was used as a loading control. B P69, DU145, and PC3 cells were treated with Sodwanone A (Sod. A) at 20 μM for 48h in hypoxia (Hx 1%). Cell lysates were analyzed by immunoblotting for HIF-1α. Tubulin was used as a loading control. C , P69, DU145, and PC3 cells were treated with Yardenone 2 (Yard. 2) at 20 μM for 48h in hypoxia (Hx 1%). Cell lysates were analyzed by immunoblotting for HIF-1α. Tubulin was used as a loading control. D, Immunofluorescence labeling and merge images showing the nuclear localization of HIF-1α (in green) and DAPI (in blue) in PC3 cells treated with Sod. A and Yard. 2 at 20 μM for 48h in hypoxia (Hx 1%)

Article Snippet: The prostate cancer DU145 (HTB-81) and PC3 (CRL-1435) cell lines, were purchased from the American Tissue Culture Collection and grown in the Dulbecco's modified eagle medium (DMEM) with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin antibiotics.

Techniques: Western Blot, Control, Immunofluorescence, Labeling

Journal: Cellular & Molecular Biology Letters

Article Title: The marine-derived HIF-1α inhibitor, Yardenone 2, reduces prostate cancer cell proliferation by targeting HIF-1 target genes

doi: 10.1186/s11658-024-00617-2

Figure Lengend Snippet: Impact of the Sodwanone A (Sod. A) and Yardenone 2 (Yard. 2) on cell proliferation and viability. A and B P69, DU145, PC3, and 786-O cells were seeded at the same density and incubated in normoxia (Nx) for 48 h in the absence (Ctl) or presence of 20 µM of Sod. A, and Yard. 2. Cell proliferation ( A ) and cell viability ( B ) were measured using an ADAM cell counter. C and D P69, DU145, PC3, and 786-O cells were seeded at the same density and incubated in hypoxia (Hx 1%) for 48 h in the absence (Ctl) or presence of 20 µM of Sod. A, and Yard. 2. Cell proliferation ( C ) and cell viability ( D ) were measured using an ADAM cell counter. ( E and F ) PC3 cells were seeded at the same density and incubated in hypoxia (Hx 1%) for 24, 48, 72, and 96 h in the absence (Ctl) or presence of 20 µM of Yard. 2. Cell proliferation ( E ) and cell viability ( F ) were measured using an ADAM cell counter. G , Clonogenic assay of P69, DU145, PC3, and 786-O cells. Cell lines were seeded at the same density and incubated in Hx 1% O 2 (Hx 1%) for 7 days (7d) in the absence (Ctl) or presence of Yard. 2 at 20 µM. H Top, Three-dimensional structures obtained from confocal image series using IMARIS software; scale bars = 200 µm. MSK-PC3 organoids have been treated for 15 days in the absence or presence of Yard. 2 (40 µM) every 3 days. Bottom, Quantification of cell area (pixels) at day 15. The 2-way ANOVA is representative of at least five different organoids. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0005. Data represent the mean ± SD of experiments performed at least three times

Article Snippet: The prostate cancer DU145 (HTB-81) and PC3 (CRL-1435) cell lines, were purchased from the American Tissue Culture Collection and grown in the Dulbecco's modified eagle medium (DMEM) with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin antibiotics.

Techniques: Incubation, Clonogenic Assay, Software

Journal: Nature Communications

Article Title: Small molecule in situ resin capture provides a compound first approach to natural product discovery

doi: 10.1038/s41467-024-49367-x

Figure Lengend Snippet: A Principal component analysis of high dimensional phenotypic profiles of 1 – 3 (red, green, yellow, respectively), reference compounds (gray), and DMSO (blue) in nine diverse cell lines: AsPC 1 (human pancreas), A549 (lung), DU145 (prostate), HCT116 (colon), HEPG2 (hepatoma), OVCAR4 (ovarian), U87MG (glioblastoma), WM164 (melanoma), 786-O (renal). DMSO (dimethyl sulfoxide). B Bioactivity expressed as significance (−log 2 p val) across all cell lines. Responses above the dashed line are considered significant ( p < 10 −6 ). P values for compound-to-DMSO distances were calculated based on the empirical null distribution of DMSO-DMSO distances (one-sided, no adjustments for multiple comparisons; Methods). C Representative Ca 2+ transient traces of DMSO (top) and 1 (bottom). D Scatter plot showing beating frequency and mean peak amplitude of 1 (red), DMSO (blue), and anti-arrhythmic and anti-cancer drugs (gray).

Article Snippet: Lung (A549, #CCL-185), prostatic (DU145, #HTB-81), renal (786-O, #CRL-1932), and pancreatic (AsPC1, #CRL-1682) carcinoma cells were purchased from ATCC (Manassas, VA).

Techniques:

Journal: Molecules

Article Title: Novel Antineoplastic Inducers of Mitochondrial Apoptosis in Human Cancer Cells

doi: 10.3390/molecules29040914

Figure Lengend Snippet: Compound 1 ( PT162 , NSC 796018 ) and compound 3 (PT167, NSC 799315) induce cytochrome c translocation from mitochondria into the cytosol, a hallmark of the intrinsic pathway of apoptosis, whereas compound 2 ( PT166 , NSC 750423 ) fails to induce apoptotic cytochrome c translocation. The cytochrome c assays (mitochondrial and cytosolic) were performed with compounds 1 , 2, and 3 in the human prostate cancer cell lines PC-3 and DU-145. The protein content for each cellular fraction was calculated to determine the mass content of cytochrome c in each fraction (pg cytochrome c /mg total protein). The mass content of cytochrome c (pg cytochrome c /mg total protein) was normalized towards the corresponding blank DMSO control and is given in percent (%) of blank DMSO control. Each experiment was representative of the effects of compound 1 , compound 2 , and compound 3 in the human prostate cancer cell lines PC-3 and DU-145. ( A ) Experiment compound 1 in PC-3 cells. ( B ) Experiment compound 1 in DU-145 cells. ( C ) Experiment compound 2 in PC-3 cells. ( D ) Experiment compound 2 in DU-145 cells. ( E ) Experiment compound 3 in PC-3 cells. ( F ) Experiment compound 3 in DU-145 cells.

Article Snippet: The metastatic prostate cancer cell lines PC-3 (CRL-1435™) (bone metastasis, adenocarcinoma) and DU-145 (HTB-81™) (brain metastasis, adenocarcinoma) were retrieved from American Type Culture Collection (ATCC ® , Manassas, VA, USA).

Techniques: Translocation Assay

Journal: Advanced Science

Article Title: Dampened Regulatory Circuitry of TEAD1/ITGA1/ITGA2 Promotes TGFβ1 Signaling to Orchestrate Prostate Cancer Progression

doi: 10.1002/advs.202305547

Figure Lengend Snippet: Loss of α1‐ or α2‐integrins leads to different but synergistic phenotypes in prostate epithelial cells. a) Level of α1‐ and α2‐integrins in normal (RWPE1) and PCa (DuCaP, PC‐3, VCaP, DU145, 22Rv1) epithelial cells. The data shows the mean value from three independent experiments, analyzed with One‐way ANOVA. b) RWPE1‐WT, RWPE1‐α1KO, RWPE1‐α2KO and RWPE1α1α2dKO cell lysates were analyzed for the expression levels of α1‐ and α2‐integrins. β‐tubulin was used as a loading control. The data shows the mean value from three independent experiments, analyzed with One‐way ANOVA. c ) RWPE1‐WT, RWPE1‐α1KO, RWPE1‐α2KO and RWPE1‐α1α2dKO cells grown on glass coverslips for 2 days were imaged using phase contrast microscopy. Scale bar = 10 µm. d) Proliferation of the indicated RWPE1 cell lines were analyzed using an XTT assay, analyzed with Two‐way ANOVA. The data shows mean ±SD from three independent experiments performed in triplicates. e) Migration of the different RWPE1 variants was analyzed using the IncuCyteS3 Scratch Wound module. f) A plot showing the wound‐closure dynamics of the indicated RWPE1 cell lines. The mean ± SD from a representative assay with 3 replicates is plotted in the graph, analyzed with Two‐way ANOVA. The assay was repeated thrice with similar results. g) Phase contrast microscopy images of the indicated RWPE1 variants grown in 3D Matrigel for 7 days. Scale bar = 50 µm. h) Quantitation of the 3D morphology analysis of RWPE1‐WT, ‐α1KO and α2KO cell lines. RWPE1‐α1α2dKO cells formed interconnected multicellular networks and could not be analyzed. Cysts were classified into 4 categories: cysts with a central hollow lumen, multilumen cysts, cysts with individual cells in the lumen and solid cysts with no visible lumen. The data shows the mean ± SD from three independent experiments in which at least 100 cysts per sample was analyzed. Statistical significance is indicated by asterisks: ∗ = p < 0.05; ∗∗ = p < 0.01; ∗∗∗ = p < 0.001.

Article Snippet: PCa cell line PC‐3 (CRl‐1435), VCap (CRL‐2876), DuCap (CVCL_2025), 22Rv1 (CRL‐2505), LNCaP (CRL‐1740) and DU145 (HTB‐81) were purchased from ATCC.

Techniques: Expressing, Microscopy, XTT Assay, Migration, Quantitation Assay

Journal: Advanced Science

Article Title: Dampened Regulatory Circuitry of TEAD1/ITGA1/ITGA2 Promotes TGFβ1 Signaling to Orchestrate Prostate Cancer Progression

doi: 10.1002/advs.202305547

Figure Lengend Snippet: TEAD1 is highly co‐expressed with ITGA1 and ITGA2 and is downregulated during PCa development and progression. a) TEAD1 expression levels are downregulated upon copy loss/del in PCa. The P value was examined by the Mann‐Whitney U test. b) PCa patients with TEAD1 copy number loss/del show higher risks for biochemical recurrence. P ‐value was assessed by the log‐rank test. c,d) TEAD1 expression levels are significantly decreased upon PCa tumor progression to metastasis. P values were determined by Kruskal‐Wallis H test. e–h) TEAD1 downregulation correlates with PCa tumor progression to high tumor stage e), Gleason score f), lymph node metastasis g) and PSA levels h). P values were determined by Kruskal‐Wallis H test or Mann‐Whitney U test g). i–k) Lower TEAD1 expression levels in PCa patients are associated with higher risks for biochemical recurrence i,j) and metastasis k). P ‐values were assessed by the log‐rank test. l) Western blot analysis of the TEAD1 expression in a benign (RWPE‐1) and malignant (DU145, PC3, LnCap, VCap) epithelial prostate cell lines. The blot is representative of three independent experiments with similar results, analyzed with One‐way ANOVA. m) RWPE1‐WT and RWPE1‐TEAD1KO cells grown on glass coverslips for 2 days were imaged using phase contrast microscopy. Scale bar = 10 µm. n) Cell proliferation analysis of RWPE1 and RWPE1‐TEAD1KO cells was done using XTT assay. The data shows mean ±SD from three independent experiments performed in triplicates, analyzed with Two‐way ANOVA. o) Cell migration analysis using the IncuCyte S3 scratch wound module. The plot shows the mean ± SD from a representative assay done in triplicate, analyzed with Two‐way ANOVA. The assay was done three times with similar results. ∗ = p < 0.05; ∗∗ = p < 0.01; ∗∗∗ = p < 0.001.

Article Snippet: PCa cell line PC‐3 (CRl‐1435), VCap (CRL‐2876), DuCap (CVCL_2025), 22Rv1 (CRL‐2505), LNCaP (CRL‐1740) and DU145 (HTB‐81) were purchased from ATCC.

Techniques: Expressing, MANN-WHITNEY, Western Blot, Microscopy, XTT Assay, Migration