Journal: Scientific Reports

Article Title: Resveratrol suppresses serum-induced vasculogenic mimicry through impairing the EphA2/twist-VE-cadherin/AKT pathway in human prostate cancer PC-3 cells

doi: 10.1038/s41598-022-24414-z

Figure Lengend Snippet: Effect of resveratrol on the cell viability against prostate cancer cells. ( A ) Chemical structure of resveratrol. PC-3 ( B) or DU145 ( C ) cells were treated with various concentrations of RES for 24 h followed by the MTT assay. Data present as the means ± SD of triplicate determinations.

Article Snippet: Resveratrol (RES) (Purity: ≥ 99% as determined by HPLC, Fig. A), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), primary antibody for β-actin and propidium iodide (PI) solution were obtained from Sigma-Aldrich (St Louis, MO, USA).

Techniques: MTT Assay

Journal: Scientific Reports

Article Title: Resveratrol suppresses serum-induced vasculogenic mimicry through impairing the EphA2/twist-VE-cadherin/AKT pathway in human prostate cancer PC-3 cells

doi: 10.1038/s41598-022-24414-z

Figure Lengend Snippet: Resveratrol suppresses serum-induced invasion in prostate cancer cells. PC-3 ( A ) or DU145 cells ( B ) with RES seeded on a matrigel-coated up chamber of transwell and serum was filled in a down chamber. After a 24 h-incubation, the cells were fixed and stained. The images were taken with an inverted microscope at 200 × magnification. Scale bar = 100 μm. The number of invading cells was counted and expressed as a percentage of the untreated control group. Data present as the mean ± SD of three independent experiments by analysis of Student’s t- test. *** p < 0.001 versus untreated control; ## p < 0.01 and ### p < 0.001 versus serum-treated control.

Article Snippet: Resveratrol (RES) (Purity: ≥ 99% as determined by HPLC, Fig. A), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), primary antibody for β-actin and propidium iodide (PI) solution were obtained from Sigma-Aldrich (St Louis, MO, USA).

Techniques: Incubation, Staining, Inverted Microscopy

Journal: Scientific Reports

Article Title: Resveratrol suppresses serum-induced vasculogenic mimicry through impairing the EphA2/twist-VE-cadherin/AKT pathway in human prostate cancer PC-3 cells

doi: 10.1038/s41598-022-24414-z

Figure Lengend Snippet: Resveratrol decreases serum-induced VM tube formation in prostate cancer cells. PC-3 ( A ) or DU145 cells ( B ) were seeded on a matrigel-coated a 24-well plate and treated with serum with or without RES. After a 16 h-incubation, the images were taken with an inverted microscope at 40 × magnification. Scale bar = 250 μm. The number of VM tube formation was counted and expressed as a percentage of the untreated control group. Data present as the mean ± SD of three independent experiments by analysis of Student’s t- test. *** p < 0.001 versus untreated control; ### p < 0.001 versus serum-treated control.

Article Snippet: Resveratrol (RES) (Purity: ≥ 99% as determined by HPLC, Fig. A), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), primary antibody for β-actin and propidium iodide (PI) solution were obtained from Sigma-Aldrich (St Louis, MO, USA).

Techniques: Incubation, Inverted Microscopy

Journal: Scientific Reports

Article Title: Resveratrol suppresses serum-induced vasculogenic mimicry through impairing the EphA2/twist-VE-cadherin/AKT pathway in human prostate cancer PC-3 cells

doi: 10.1038/s41598-022-24414-z

Figure Lengend Snippet: Resveratrol inhibits serum-induced the activation of EphA2 in PC-3 cells. Cells were treated with serum with or without RES for 24 h. ( A ) The same amount of proteins (25–30 μg) were analyzed by Western blot using the phospho-EphA2 and EphA2 antibodies. As a loading control, β-actin was used. ( B ) The bands of proteins were quantified. Data present as the means ± SD of three independent experiments by analysis of Student’s t -test. *** p < 0.001 versus untreated control; # p < 0.05 and ## p < 0.01 versus serum-treated control.

Article Snippet: Resveratrol (RES) (Purity: ≥ 99% as determined by HPLC, Fig. A), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), primary antibody for β-actin and propidium iodide (PI) solution were obtained from Sigma-Aldrich (St Louis, MO, USA).

Techniques: Activation Assay, Western Blot

Journal: Scientific Reports

Article Title: Resveratrol suppresses serum-induced vasculogenic mimicry through impairing the EphA2/twist-VE-cadherin/AKT pathway in human prostate cancer PC-3 cells

doi: 10.1038/s41598-022-24414-z

Figure Lengend Snippet: Resveratrol down-regulates serum-induced VE-cadherin expression through decreasing the nuclear twist expression in PC-3 cells. Cells were treated with serum with or without RES for 24 h. ( A ) The same amount of proteins (25–30 μg) were analyzed by Western blot using the VE-cadherin antibody. As a loading control, β-actin was used. ( B )) The mRNA levels were analyzed by RT-PCR using the VE-cadherin primer. As a loading control, β-actin primer was used. ( C ) The same amount of proteins (25–30 μg) were analyzed by Western blot using the twist antibody. β-actin was used as a control. Data present as the means ± SD of three independent experiments by analysis of Student’s t- test. * p < 0.05 and *** p < 0.001 versus untreated control; ## p < 0.01 and ### p < 0.001 versus serum-treated control. ( D ) The serum-treated cells with or without RES for 24 h were fixed, permeabilized and blocked. After probing with Twist antibody followed by incubation with fluorescein isothiocyanate (FITC)-conjugated secondary antibody, the cells were counterstained with propidium iodide (PI). The images were taken with a fluorescence microscope at 400 × . Scale bar = 50 μm.

Article Snippet: Resveratrol (RES) (Purity: ≥ 99% as determined by HPLC, Fig. A), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), primary antibody for β-actin and propidium iodide (PI) solution were obtained from Sigma-Aldrich (St Louis, MO, USA).

Techniques: Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction, Incubation, Fluorescence, Microscopy

Journal: Scientific Reports

Article Title: Resveratrol suppresses serum-induced vasculogenic mimicry through impairing the EphA2/twist-VE-cadherin/AKT pathway in human prostate cancer PC-3 cells

doi: 10.1038/s41598-022-24414-z

Figure Lengend Snippet: Resveratrol inactivates serum-induced the AKT signaling pathway in PC-3 cells. Cells were treated with serum with or without RES for 24 h. The same amount of proteins (25–30 μg) were analyzed by Western blot using the phospho-AKT, AKT ( A ), MMP-2 ( B ) and LAMC2 ( D ) antibodies. As a loading control, β-actin was used. ( C ) Gelatin zymography was performed using the CM. Data present as the means ± SD of three independent experiments by analysis of Student’s t- test. *** p < 0.001 versus untreated control; ## p < 0.01 and ### p < 0.001 versus serum-treated control.

Article Snippet: Resveratrol (RES) (Purity: ≥ 99% as determined by HPLC, Fig. A), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), primary antibody for β-actin and propidium iodide (PI) solution were obtained from Sigma-Aldrich (St Louis, MO, USA).

Techniques: Western Blot, Zymography