du145 pc cell line  (ATCC)

Journal: Oncotarget

Article Title: Development of a multiplex assay to assess activated p300/CBP in circulating prostate tumor cells.

doi: 10.18632/oncotarget.28477

Figure Lengend Snippet: Figure 1: Optimization of circulating tumor cells (CTC) staining and validation of p300 activity. Prostate cancer cell lines LNCaP and Du145 cells were spiked into whole blood and captured using an EpCAM antibody and stained with acetyl-p300, total p300, acetyl-H3K18 and SIRT2 epigenetic biomarkers Cells of exclusion (not captured with EpCAM) being stained with CD11b, CD66b, CD45 and CD34 showed no positive staining. (A) Cells were stained (LNCaP shown) and imaged with a fluorescence microscope (Magnification X200). Cytokeratin (CK) is used to identify intact cells and Hoeschst staining employed for evaluating cell nuclei. (B) Cells were treated with 0.3 µM of A-485 for 24 hours then harvested for staining and analysis. A-485 is a selective catalytic inhibitor for p300 histone acetyltransferase. Staining for each channel is demonstrated. A reduction in acetyl-P300 and acetyl-H3K18 (left shift) is noted after inhibitor treatment. Assays were performed in duplicate with excellent reproducibility. (C) Western blot confirms the reduction in acetylated p300 and its target, acetylated H3k18 by A-485 seen on fluorescent staining. Experiments were repeated and performed in duplicate.

Article Snippet: The PC cell lines Du145 is resistant to androgen deprivation therapy, and androgen-sensitive LNCaP were obtained from the ATCC and genotyped for validity.

Techniques: Staining, Biomarker Discovery, Activity Assay, Fluorescence, Microscopy, Western Blot

Journal: The Chinese journal of physiology

Article Title: Leonurine suppresses prostate cancer growth in vitro and in vivo by regulating miR-18a-5p/SLC40A1 axis.

doi: 10.4103/0304-4920.365459

Figure Lengend Snippet: Figure 1: Leo inhibits prostate cancer cell proliferation. (a) Chemical structure of leonurine. (b) CCK‑8 assay was used to evaluate cell viability of PC3 and DU145 cells treated with various concentrations of Leo at 0 h, 24 h, 48 h, and 72 h, separately (n = 3). One‑way ANOVA with post hoc test was used to compare results from different groups. (c) Colony formation assay was performed to assess proliferation capacity of PC3 and DU145 cells treated with various concentrations of Leo. The number of colonies was quantified (n = 3). One‑way ANOVA with post hoc test was used to compare results from different groups. (d) Fluorescent staining of BrdU was performed to identify BrdU+ cell in PC3 and DU145 cells treated with various concentrations of Leo. The number of BrdU+ cells was quantified (n = 3). One‑way ANOVA with post hoc test was used to compare results from different groups. ** indicates P < 0.01, *** indicates P < 0.001 compared with the Leo (0 μM) group. SLC40A1: Solute carrier family 40 member 1, CCK‑8: Cell counting kit‑8, ANOVA: Analysis of variance, BrdU: 5-Bromo-2-deoxyuridine.

Article Snippet: PC-3 and DU145 cell lines were purchased from American Type Culture Collection (Bethesda, MD, USA).

Techniques: CCK-8 Assay, Colony Assay, Staining

Journal: The Chinese journal of physiology

Article Title: Leonurine suppresses prostate cancer growth in vitro and in vivo by regulating miR-18a-5p/SLC40A1 axis.

doi: 10.4103/0304-4920.365459

Figure Lengend Snippet: Figure 2: Leo leads to prostate cancer cell cycle arrest. (a) Flow cytometry was performed to evaluate cell cycle in PC3 and DU145 cells treated with various concentrations of Leo. The cell ratios of G1 phase, G2 phase, and S phase were quantified (n = 3). One‑way ANOVA with post hoc test was used to compare results from different groups. (b) Immunoblotting was performed to detect CDK2 and cyclin E protein expression in PC3 and DU145 cells treated with various concentrations of Leo. The grayscale value of the corresponding band was quantitated with ImageJ (n = 3). One‑way ANOVA with post hoc test was used to compare results from different groups. * indicates P < 0.05 and ** indicates P < 0.01 compared with the Leo (0 μM) group. ANOVA: Analysis of variance.

Article Snippet: PC-3 and DU145 cell lines were purchased from American Type Culture Collection (Bethesda, MD, USA).

Techniques: Flow Cytometry, Western Blot, Expressing

Journal: The Chinese journal of physiology

Article Title: Leonurine suppresses prostate cancer growth in vitro and in vivo by regulating miR-18a-5p/SLC40A1 axis.

doi: 10.4103/0304-4920.365459

Figure Lengend Snippet: Figure 3: Leo leads to prostate cancer cell apoptosis. (a) Flow cytometry was performed to evaluate the cell apoptosis ratio of PC3 and DU145 cells treated with various concentrations of Leo. The cell apoptosis ratio was quantified (n = 3). One‑way ANOVA with post hoc test was used to compare results from different groups. (b) Immunoblotting was performed to detect Bax and Bcl‑2 protein expression in PC3 and DU145 cells treated with various concentrations of Leo. The grayscale value of the corresponding band was quantitated with ImageJ (n = 3). One‑way ANOVA with post hoc test was used to compare results from different groups. * indicates P < 0.05 and ** indicates P < 0.01 compared with the Leo (0 μM) group. ANOVA: Analysis of variance.

Article Snippet: PC-3 and DU145 cell lines were purchased from American Type Culture Collection (Bethesda, MD, USA).

Techniques: Flow Cytometry, Western Blot, Expressing

Journal: The Chinese journal of physiology

Article Title: Leonurine suppresses prostate cancer growth in vitro and in vivo by regulating miR-18a-5p/SLC40A1 axis.

doi: 10.4103/0304-4920.365459

Figure Lengend Snippet: Figure 4: Leo suppresses prostate cancer cell growth depending on the upregulation of SLC40A1. (a) RT‑PCR was performed to detect miR‑18a‑5p expression in PC3 and DU145 cells treated with various concentrations of Leo (n = 3). One‑way ANOVA with post hoc test was used to compare results from different groups. (b) Immunoblotting was performed to detect SLC40A1 protein expression in PC3 and DU145 cells treated with various concentrations of Leo. The grayscale value of the corresponding band was quantitated with ImageJ (n = 3). One‑way ANOVA with post hoc test was used to compare results from different groups. ** indicates P < 0.01 compared with the Leo (0 μM) group. (c) Colony formation assay was performed to assess the proliferation capacity of PC3 and DU145 cells transfected with si‑NC or si‑SLC40A1 under Leo (800 μM) treatment. The number of colonies was quantified (n = 3). One‑way ANOVA with post hoc test was used to compare results from different groups.(d) Flow cytometry was performed to evaluate the cell apoptosis ratio of PC3 and DU145 cells transfected with si‑NC or si‑SLC40A1 under Leo (800 μM) treatment. The cell apoptosis ratio was quantified (n = 3). One‑way ANOVA with post hoc test was used to compare results from different groups. (e) Flow cytometry was performed to evaluate cell cycle in PC3 and DU145 cells transfected with si‑NC or si‑SLC40A1 under Leo (800 μM) treatment. The cell ratios of G1 phase, G2 phase, and S phase were quantified (n = 3). One‑way ANOVA with post hoc test was used to compare results from different groups. ** indicates P < 0.01 compared with the control group. ##indicates P < 0.01 compared with the Leo (800 μM) + si‑NC group. ANOVA: Analysis of variance.

Article Snippet: PC-3 and DU145 cell lines were purchased from American Type Culture Collection (Bethesda, MD, USA).

Techniques: Expressing, Western Blot, Colony Assay, Transfection, Flow Cytometry, Control