Effects Of Vbe-3, A Noval Lignanoid From The Acetoacetate Extract Of Vitex Negundo Seed On The Growth And Akt Signaling Of Human Hepatocellular Carcinoma Hepg2 Cell Line

Objective:To investigate the effects of VBE-3, a noval lignanoid from the acetoacetate extract of Vitex negundo seed on the growth and Akt signal pathway in human hepatocellular carcinoma HepG2 cell line.Methods:Human hepatocellular carcinoma HepG2 cell line and human embryon liver L-02 cell line were cultured in vitro. MTT assay was used to determine the cell viability in both cell lines. The trypan blue exclusion method was used to count the number of living cells, to draw cell growth curve. Western blot was used to analyze expression of p-Akt, p-mTOR and p-ERK1/2 protein in HepG2 cells.Results:The MTT assay showed that cell viability inhibitory rate was 6.45%±0.19%,9.67%±0.56%,22.22%±0.32%,35.32%±0.54%,43.29%±0.44% and 58.18%±0.79%, respectively, after HepG2 cells treatment with VBE-3 at various concentration of 0.1,0.3,1.0,3.0,10.0,30.0μmol/L for 48h, with IC50 was 13.86μmol/L. The cell viability inhibitory rate was 1.06%±0.04%,7.65%±0.21%,15.16%±3.2%,8.73%±5.90%,22.55%±5.06% and 27.07 %±8.35%, respectively, after treatment with VBE-3 at various concentration of 0.1, 0.3,1.0,3.0,10.0,30.0μmol/L for 48h, with IC50 was 79.98μmol/L, in L-02 cell line. Using trypan blue exclusion method, the results showed VBE-3 at 10.0μmol/L had inhibitory growth of HepG2 cells, in a time-dependent manner. Western blot analysis indicated that exposure to VBE-3 or LY294002 or both at 10.0μmol/L for 24h, the phosphorylated protein level of Akt in HepG2 cells was decreased by 87.76 %±1.60%,90.00%±2.26% and 82.06%±1.79%, respectively in comparision with the control group. At same condition, the phosphorylated protein level of mTOR and ERK1/2 were down-regulated by 96.30%±1.87%,91.76%±1.34%,86.86%±2.26% and 85.83%±1.13%,91.40%±1.74%,82.53%±1.87%,respectively in comparision with the control group.Conclusion:1.VBE-3 selectively inhibite the proliferation and growth of HepG2 cells.2.The growth inhibitory effects of VBE-3 in HepG2 cell line may be involved in the down-regulation of the phosphorylated protein level of Akt,mTOR and ERK1/2.