normal prostate epithelial cell line hprec  (ATCC)

Journal: Current Issues in Molecular Biology

Article Title: Effect of Soy Isoflavone on Prostate Cancer Cell Apoptosis Through Inhibition of STAT3, ERK, and AKT

doi: 10.3390/cimb46110743

Figure Lengend Snippet: Effects of genistein on cell viability, cell morphology, and nuclear morphology. ( A ) Chem ical structure of genistein. ( B ) Cell viability was measured using an MTT assay in normal HPrEC and DU145 cells. Cells were exposed to increasing concentrations of genistein (0, 5, 10, 20, 30, 40, 50, 75, 100, and 200 μM). ( C ) Cell morphology was assessed under phase-contrast microscopy in HPrEC and DU145 cells. ( D ) Nuclear morphology was evaluated with a fluorescence microscope following DAPI (2 μg/mL) staining in HPrEC and DU145 cells. ( E ) A wound healing assay was conducted. * p < 0.05 was compared to respective controls. GEN: genistein, HPrEC: human prostate epithelial cell line, DAPI: 4′,6-diamidino-2-phenylindole, and Conc.: concentration.

Article Snippet: The human prostate epithelial cell line HPrEC and the human prostate cancer cell line DU145 were sourced from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: MTT Assay, Microscopy, Fluorescence, Staining, Wound Healing Assay, Concentration Assay

Journal: Current Issues in Molecular Biology

Article Title: Effect of Soy Isoflavone on Prostate Cancer Cell Apoptosis Through Inhibition of STAT3, ERK, and AKT

doi: 10.3390/cimb46110743

Figure Lengend Snippet: Caspase-3 activation by genistein in HPrEC and DU145 cells. ( A – C ) Cells were treated with increasing concentrations (0–100 μM) of genistein for 48 h. Caspase-3/7 activity was determined using an ApoTox-Glo TM Triplex Assay ( A ). Cell lysates were subsequently analyzed by Western blotting with antibodies against procaspase-3, cleaved caspase-3, PARP, Bax, Bcl-2, and β-actin ( B ). The percentage of apoptotic cells after annexin V-PE binding was quantified by a Muse cell analyzer ( C ). ( D – F ) Cells underwent treatment with genistein (100 μM) for 48 h in the presence or absence of Z-VAD (10 μM). Caspase-3/7 activity was determined using the ApoTox-Glo TM Triplex Assay ( D ). Cell lysates were then analyzed with Western blotting for procaspase-3, cleaved caspase-3, PARP, and β-actin ( E ). The percentage of apoptotic cells after annexin V-PE binding was determined using a Muse cell analyzer ( F ). Error bars represent mean ± S.D. from three independent experiments. * p < 0.05 was compared to respective controls. # p < 0.05 was compared to Z-VAD (−) groups. GEN: genistein, HPrEC: human prostate epithelial cell line, and Conc.: concentration.

Article Snippet: The human prostate epithelial cell line HPrEC and the human prostate cancer cell line DU145 were sourced from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Activation Assay, Activity Assay, Western Blot, Binding Assay, Concentration Assay

Journal: Current Issues in Molecular Biology

Article Title: Effect of Soy Isoflavone on Prostate Cancer Cell Apoptosis Through Inhibition of STAT3, ERK, and AKT

doi: 10.3390/cimb46110743

Figure Lengend Snippet: Effects of genistein and pan-caspase inhibitor on cell distribution at sub-G 0 /G 1 , G 1 , S, and G 2 /M phases in HPrEC and DU145 cells. Cells were treated with increasing concentrations (0–100 μM) of genistein for 48 h. Cells were treated with Z-VAD (10 μM) for 2 h prior to incubation with genistein (100 μM) for 48 h. ( A , B ) Subsequently, cells were analyzed using flow cytometry after staining with propidium iodide (20 μg/mL). Representative results were sourced from one of three independent experiments. The quantitative data are presented as mean ± S.D. * p < 0.05 was compared to respective controls. # p < 0.05 was compared to Z-VAD (−) groups. GEN: genistein, and HPrEC: human prostate epithelial cell line.

Article Snippet: The human prostate epithelial cell line HPrEC and the human prostate cancer cell line DU145 were sourced from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Incubation, Flow Cytometry, Staining

Journal: Current Issues in Molecular Biology

Article Title: Effect of Soy Isoflavone on Prostate Cancer Cell Apoptosis Through Inhibition of STAT3, ERK, and AKT

doi: 10.3390/cimb46110743

Figure Lengend Snippet: Effects of genistein and pan-caspase inhibitor on ROS and mitochondrial function in HPrEC and DU145 cells. Cells were treated with Z-VAD (10 μM) for 2 h before incubation with genistein (100 μM) for 48 h. ( A , B ) Cells were subsequently analyzed using flow cytometry after staining with propidium iodide (20 μg/mL). ( C , D ) Mitochondrial membrane potential was measured after staining the cells with rhodamine123. ( E , F ) Intracellular ATP levels. Representative results were obtained from one of three independent experiments. The quantitative data are presented as mean ± S.D. * p < 0.05 was compared to respective controls. # p < 0.05 was compared to Z-VAD (−) groups. GEN: genistein; HPrEC: human prostate epithelial cell line.

Article Snippet: The human prostate epithelial cell line HPrEC and the human prostate cancer cell line DU145 were sourced from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Incubation, Flow Cytometry, Staining, Membrane

Journal: Current Issues in Molecular Biology

Article Title: Effect of Soy Isoflavone on Prostate Cancer Cell Apoptosis Through Inhibition of STAT3, ERK, and AKT

doi: 10.3390/cimb46110743

Figure Lengend Snippet: Effects of genistein on the p53, p21, AKT, ERK, MAPK, STAT3 pathways, and VEGF protein levels in HPrEC and DU145 cells. Cells were treated with increasing concentrations (0–100 μM) of genistein for 48 h. ( A – E ) The cell lysates were subjected to Western blot analysis targeting proteins such as p53, p21, p-AKT, AKT, p-ERK, ERK, p-p38, p38, p-STAT3, STAT3, and VEGF. GEN: genistein; HPrEC: human prostate epithelial cell line.

Article Snippet: The human prostate epithelial cell line HPrEC and the human prostate cancer cell line DU145 were sourced from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Western Blot