Abstract:Objective To observe the effect of Raddeanin A on the expression of vascular endothelial growth factor(VEGF) gene in human hepatocellular carcinoma HepG-2 cells. Methods HepG-2 cells were cultured with different concentrations of Raddeanin A in vitro. Enzyme-linked immuno sorbent assay(ELISA) was used to detect VEGF level in cell culture supernatant after HepG-2 cells treated with different concentration of Raddeanin A for 24 h. Reverse transcription-polymerase chain reaction(RT-PCR) and protein immunoblot (western blot) were used to detect the relative expression level of messenger ribonucleic acid (mRNA) and protein (the ratio of gray scale value of the positive band to the β-actin) of VEGF in HepG-2 cells respectively. Results (1) After HepG-2 cells were treated by different concentrations (5, 10 and 20 μg/ml) of Raddeanin A, the level of VEGF protein in cell culture supernatant decreased in a dose dependent manner(P=0.000). (2) The relative expression level of VEGF mRNA decreased in a dose dependent manner after HepG-2 cells treated with 0, 5, 10 and 20 μg/ml Raddeanin A for 24 h (0.96 ± 0.07 vs 0.75 ± 0.09 vs 0.55 ± 0.10 vs 0.32 ± 0.11, P=0.000). The relative expression level of VEGF mRNA decreased in a time dependent manner after HepG-2 cells treated with 10 μg/ml Raddeanin A for 0, 6, 12 and 24 h (0.77±0.05 vs 0.53±0.09 vs 0.32±0.12 vs 0.15±0.04, P=0.000). (3) The relative expression level of VEGF protein decreased in a dose dependent manner after HepG-2 cells treated with 0, 5, 10 and 20 μg/ml Raddeanin A for 24 h (2. 08±0.28 vs 1. 43±0.22 vs 0.72±0.15 vs 0.17±0.10, P=0.000). The relative expression level of VEGF protein decreased in a time dependent manner after HepG-2 cells treated with 10 μg/ml Raddeanin A for 0, 6, 12 and 24 h (2. 47±0.05 vs 1. 36±0.02 vs 0.30±0.02 vs 0.07±0.02, P=0.000). Conclusion Raddeanin A could down-regulate the expression of VEGF gene in HepG-2 cells, which may be one of the important mechanisms to inhibit the growth and proliferation of liver cancer cells.