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: Journal of Immunology Research

Article Title: Combination Blockade of the IL6R/STAT-3 Axis with TIGIT and Its Impact on the Functional Activity of NK Cells against Prostate Cancer Cells

doi: 10.1155/2022/1810804

Figure Lengend Snippet: Advanced prostate cancer cells secrete a higher concentration of growth factors and cytokines according to stage progression. The cells were cultured at different times, the supernatants from PCa lines were collected, and 13 cytokines and 10 growth factors were evaluated using LEGENDplex™ technology. (a, b) The concentration of growth factors and cytokines from the 22Rv1, LNCaP, and DU145 supernatant was evaluated at 12, 24, and 48 h. (c,e) The concentration of soluble molecules increases, conforming with the advances of stage progression; the highest VEGF, M-CSF, CXCL8, and IL-6 values were observed in the DU145 cell line. Data are shown as the mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 (ANOVA with Tukey's multiple comparison test).

Article Snippet: The prostate cancer (PCa) cell lines DU145/ATCC® HTB-81™ (metastatic tumor and castration-resistant), LNCaP clone FGC/ATCC® CRL1740™ (metastatic tumor and androgen-dependent), and 22Rv1/ATCC® CRL2505™ (non-metastatic tumor) were obtained from the ATCC® (Manassas, VA, USA).

Techniques: Concentration Assay, Cell Culture

Journal: Journal of Immunology Research

Article Title: Combination Blockade of the IL6R/STAT-3 Axis with TIGIT and Its Impact on the Functional Activity of NK Cells against Prostate Cancer Cells

doi: 10.1155/2022/1810804

Figure Lengend Snippet: The expression of CD155 is increased in DU145 cells. The cells were cultured, and the expression of the ligands was subsequently evaluated by flow cytometry. (a) Ten thousand events from the region of the live cells were implemented for ligand evaluation. (b, c) The average percentage expression and (d) MFI of the ligands in the three PCa cell lines are shown. All experiments were repeated at least three times. Data are shown as the mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 (ANOVA with Tukey's multiple comparison test).

Article Snippet: The prostate cancer (PCa) cell lines DU145/ATCC® HTB-81™ (metastatic tumor and castration-resistant), LNCaP clone FGC/ATCC® CRL1740™ (metastatic tumor and androgen-dependent), and 22Rv1/ATCC® CRL2505™ (non-metastatic tumor) were obtained from the ATCC® (Manassas, VA, USA).

Techniques: Expressing, Cell Culture, Flow Cytometry

Journal: Journal of Immunology Research

Article Title: Combination Blockade of the IL6R/STAT-3 Axis with TIGIT and Its Impact on the Functional Activity of NK Cells against Prostate Cancer Cells

doi: 10.1155/2022/1810804

Figure Lengend Snippet: Stattic and tocilizumab combination increase pSTAT-3 dephosphorylation in DU145 cells. The cells were cultured with the different treatments according to the corresponding groups for 24 h. (a) IL6R/STAT-3 axis expression was compared between PCa cell lines using 50 μ g of protein by Western blot; the presence of constitutive pSTAT-3 expression was shown in the DU145 line only; protein expression was then verified by in-cell Western. (b) Use of treatment with Stt causes a decrease in metabolic activity with concentrations greater than 5 μ M. (c) The effective Stt decrease is only shown at concentrations above the IC50. (d) The combined treatments with Tcz allow a lower Stt concentration than the IC50 with greater efficiency in the dephosphorylation of pSTAT-3. All experiments were repeated at least three times. Data are shown as the mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001; (unpaired t -test) against the basal group. Stt: stattic; Tcz: tocilizumab.

Article Snippet: The prostate cancer (PCa) cell lines DU145/ATCC® HTB-81™ (metastatic tumor and castration-resistant), LNCaP clone FGC/ATCC® CRL1740™ (metastatic tumor and androgen-dependent), and 22Rv1/ATCC® CRL2505™ (non-metastatic tumor) were obtained from the ATCC® (Manassas, VA, USA).

Techniques: De-Phosphorylation Assay, Cell Culture, Expressing, Western Blot, In-Cell ELISA, Activity Assay, Concentration Assay

Journal: Journal of Immunology Research

Article Title: Combination Blockade of the IL6R/STAT-3 Axis with TIGIT and Its Impact on the Functional Activity of NK Cells against Prostate Cancer Cells

doi: 10.1155/2022/1810804

Figure Lengend Snippet: Stattic + Anti-TIGIT + Tocilizumab increase the cytotoxicity of NK-92 cells against DU145 prostate cancer cells. (a) Significant decrease in KT50 in the Stt + Anti-TIGIT + Tcz treated groups compared to the basal group. (b) A significant increase was observed in the percentage of cytolysis in the groups treated with Stt + Anti-TIGIT + Tcz compared with the basal group observed at 4 and 24 h of coculture in both ranges E : T. All experiments were repeated at least three times. Data are shown as the mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 (ANOVA with Bonferroni multiple comparison test). Stt: stattic; Tcz: tocilizumab.

Article Snippet: The prostate cancer (PCa) cell lines DU145/ATCC® HTB-81™ (metastatic tumor and castration-resistant), LNCaP clone FGC/ATCC® CRL1740™ (metastatic tumor and androgen-dependent), and 22Rv1/ATCC® CRL2505™ (non-metastatic tumor) were obtained from the ATCC® (Manassas, VA, USA).

Techniques:

Journal: Molecular Biomedicine

Article Title: Knocking down claudin receptors leads to a decrease in prostate cancer cell migration, cell growth, cell viability and clonogenic cell survival

doi: 10.1186/s43556-021-00053-0

Figure Lengend Snippet: Cldn expression in human prostate cells and tissue specimens. a The levels of Cldn3 and Cldn4 were higher in metastatic human prostate cancer (PC3, LNCaP, and DU145) cells compared to benign prostate (RWPE-1) cells. Immunohistochemistry was performed on ( b ) human benign prostatic hyperplasia specimen, ( c ) human high-grade prostate cancer specimen 1 (Gleason grade group 4 +), and ( d ) human high-grade prostate cancer specimen 2 (Gleason grade group 4 +) for Cldn4 (brown). Representative H&E and IHC staining are shown at 100× and 400×. Scale bars = 50 μM

Article Snippet: PC3 (CRL-1435), LNCaP (CRL-1740), and DU145 (HTB-81) prostate cancer cells were acquired from the American Type Culture Collection (ATCC).

Techniques: Expressing, Immunohistochemistry

Journal: Molecular Biomedicine

Article Title: Knocking down claudin receptors leads to a decrease in prostate cancer cell migration, cell growth, cell viability and clonogenic cell survival

doi: 10.1186/s43556-021-00053-0

Figure Lengend Snippet: Knock down of Cldn3 and Cldn4 in human prostate cancer cells. Western blots showing the effects of Cldn3 siRNAs (siCldn3) and Cldn4 siRNAs (siCldn4) on the expression of ( a ) Cldn3 and ( b ) Cldn4 respectively, in human PC3 and LNCaP prostate cancer cells after 72 h. Four Cldn3 and Cldn4 siRNAs were tested. c A heatmap demonstrates the percent knockdown (i.e., Color scale is used where white represents a 100% knockdown of protein expression and dark blue represents a 0% knockdown of protein expression) for each siRNA target sequence for PC3 and LNCaP prostate cancer cells. d Relative Cldn3 protein levels in LNCaP prostate cancer cells after 2 weeks of siCldn3 and siCldn4 treatment

Article Snippet: PC3 (CRL-1435), LNCaP (CRL-1740), and DU145 (HTB-81) prostate cancer cells were acquired from the American Type Culture Collection (ATCC).

Techniques: Western Blot, Expressing, Sequencing

Journal: Molecular Biomedicine

Article Title: Knocking down claudin receptors leads to a decrease in prostate cancer cell migration, cell growth, cell viability and clonogenic cell survival

doi: 10.1186/s43556-021-00053-0

Figure Lengend Snippet: Assessment of cell growth upon siCldn treatment. Cell growth of ( a ) LNCaP and ( b ) PC3 human prostate cancer cells following siCldn3 or siCldn4 treatment relative to growth following siSC. Data are shown as mean ± SD of 3 to 4 independent experiments. * represents p < 0.05 and *** represents p < 0.001

Article Snippet: PC3 (CRL-1435), LNCaP (CRL-1740), and DU145 (HTB-81) prostate cancer cells were acquired from the American Type Culture Collection (ATCC).

Techniques:

Journal: Molecular Biomedicine

Article Title: Knocking down claudin receptors leads to a decrease in prostate cancer cell migration, cell growth, cell viability and clonogenic cell survival

doi: 10.1186/s43556-021-00053-0

Figure Lengend Snippet: The effects of knocking down Cldn3 and Cldn4 on cell viability. Cell viability was assessed on human ( a ) LNCaP or ( b ) PC3 prostate cancer cells following siCldn3 or siCldn4 treatment. c Crystal violet staining was performed to visualize viable cells. Data are shown as mean ± SD of 3 to 4 independent experiments. *** represents p < 0.001 and **** represents p < 0.0001

Article Snippet: PC3 (CRL-1435), LNCaP (CRL-1740), and DU145 (HTB-81) prostate cancer cells were acquired from the American Type Culture Collection (ATCC).

Techniques: Staining

Journal: Molecular Biomedicine

Article Title: Knocking down claudin receptors leads to a decrease in prostate cancer cell migration, cell growth, cell viability and clonogenic cell survival

doi: 10.1186/s43556-021-00053-0

Figure Lengend Snippet: Cell migration of prostate cancer cells treated with siCldn. The percent difference of ( a ) LNCaP and ( b ) PC3 prostate cancer cell scratch coverage between treatments (siCldn3, orange line and siCldn4, blue line) and control (siSC, black line) was determined at 6, 12, and 18 h. Data are shown as mean ± SD of 3 to 4 independent experiments where * represents p < 0.05

Article Snippet: PC3 (CRL-1435), LNCaP (CRL-1740), and DU145 (HTB-81) prostate cancer cells were acquired from the American Type Culture Collection (ATCC).

Techniques: Migration