Oridonin enhances in vitro anticancer effects of lentinan in SMMC-7721 human hepatoma cells through apoptotic genes

The aim of the present study was to determine the ability of oridonin to enhance the anticancer activity of lentinan (LNT) in SMMC-7721 human hepatoma cells in vitro by using various techniques, including MTT, flow cytometry, quantitative PCR and western blot assays. The results demonstrated that 20 mu g/ml was a non-toxic concentration of oridonin for L02 normal liver cells and SMMC-7721 cells, while 0-200 mu g/ml of LNT only had anti-proliferative effects on SMMC-7721 cells. LNT at 100 and 200 mu g/ml inhibited the growth of SMMC-7721 cells by 22.8 and 60.0%, respectively, and after addition of 20 mu g/ml oridonin, the inhibitory rate of 100 and 200 mu g/ml LNT was increased to 47.2 and 80.7%, respectively. Oridonin (20 mu g/ml) + LNT (200 mu g/ml)-treated SMMC-7721 cells showed the highest apoptotic rate, which was 40.5 +/- 2.5%, which was higher than that of cells treated with LNT only. LNT raised the mRNA and protein expression of caspase-3, -8 and -9 as well as B-cell lymphoma 2 (Bcl-2)-associated X protein, p53 and p21, while reducing the expression of Bcl-2, Bcl extra large protein, epidermal growth factor (EGF) and EGF receptor expression in SMMC-7721 cells as compared to that in control cells. Treatment with 20 mu g/ml oridonin and 200 mu g/ml LNT increased these changes of gene expression. From the obtained results, it may be concluded that oridonin raised the in vitro anti-cancer effects of LNT in SMMC-7721 cells. Oridonin may also be used as a sensitizing agent to increase the anticancer activity of LNT in vivo.