Resveratrol promotes regression of renal carcinoma cells via a renin-angiotensin system suppression-dependent mechanism

Figures

• 

  Figure 1.. Resveratrol suppressed renal carcinoma cell proliferation. (A) Cell growth, as detected by Cell Counting Kit-8 assay, was suppressed in a dose- and time-dependent manner subsequent to resveratrol treatment. (B) Resveratrol inhibited proliferation in ACHN and A498 cells at 24 h, which was visualized by microscopy (magnification, ×100). (C) Representative pictures and quantification of resveratrol-induced colony formation suppression in renal carcinoma cells followed 6 h resveratrol treatment. *P<0.05 and **P<0.01, experimental vs. control groups.

• 

  Figure 2.. Resveratrol induced apoptosis in renal carcinoma cells. (A) Cytotoxicity of resveratrol was determined by a lactate dehydrogenase assay. (B) Caspase 3/7 activity subsequent to resveratrol treatment was determined by a Caspase-Glo 3/7 assay. (C) Resveratrol-induced apoptosis in renal carcinoma cells with different concentrations (0, 50, 100 and 200 µM) for 24 h, which was analyzed by flow cytometry. (D) Cells were treated with resveratrol (0, 50, 100 and 200 µM) for 24 h, and the levels of caspase 9, Bax and Bcl-2 were detected by western blotting. The data showed that resveratrol regulated apoptotic proteins in renal carcinoma cells in a dose-dependent manner. **P<0.01, experimental vs. control groups. JS-K, O2-(2,4-dinitrophenyl)1-[(4-ethoxyxarbonyl) piperazin-1-yl]diazen-1-ium-1,2-diolate; FITC, fluorescein isothiocyanate; Bcl-2, B-cell lymphoma 2; Bax, bcl-2-like protein 4; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

• 

  Figure 3.. Resveratrol suppressed intracellular AngII, AT1R, VEGF and COX-2 levels in a dose-dependent manner. Subsequent to renal carcinoma cells being treated with resveratrol (0, 50, 100 and 200 µM) for 24 h, the intracellular levels of (A) AngII, (B) AT1R, (C) VEGF and (D) COX-2 were determined. *P<0.05 and **P<0.01, experimental vs. control groups. AngII, angiotensin II; AT1R, AngII type 1 receptor; VEGF, vascular endothelial growth factor; COX-2, cyclooxygenase-2.

• 

  Figure 4.. Resveratrol decreased AngII, AT1R, VEGF and COX-2 levels in culture medium for renal carcinoma cells. The levels of (A) AngII, (B) AT1R, (C) VEGF and (D) COX-2 in the culture medium for renal carcinoma cells subsequent to being cultured for 24 h were determined using enzyme linked immunosorbent assays. *P<0.05 and **P<0.01, experimental vs. control groups. AngII, angiotensin II; AT1R, AngII type 1 receptor; VEGF, vascular endothelial growth factor; COX-2, cyclooxygenase-2.

• 

  Figure 5.. AT1R and VEGF reversed resveratrol-induced cell proliferation inhibition and apoptosis. (A) Resveratrol-induced cell proliferation inhibition in renal carcinoma cells with or without (a) AngII (2 pg/ml), (b) AT1R (5 pg/ml), (c) VEGF (2 pg/ml) and (d) COX-2 (0.5 ng/ml). (B) Apoptosis of (a) ACHN and (b) A498 cells was analyzed according to flow cytometry distributions with or without AngII (2 pg/ml), AT1R (5 pg/ml), VEGF (2 pg/ml) and COX-2 (0.5 ng/ml). *P<0.05, experimental vs. control groups. AngII, angiotensin II; AT1R, AngII type 1 receptor; VEGF, vascular endothelial growth factor; COX-2, cyclooxygenase-2; FITC, fluorescein isothiocyanate.