Inhibited Mechanism Of Total Saponins From Physalis Alkekengi L. Var. Franchetii Makino

[Objective] To explore the anti-cancer molecular and protein mechanism of TSP,the inhibitory effects of the total saponins from Physalis alkekengi L. Var. frawcheti(Mast.)Makino(TSP) on the proliferation of SMMC-7721 human hepatoma cells.[Methods] 1.The proliferous inhibitory effect of the TSP on SMMC-7721 human hepatoma cells was treated with MTT assay. The results showed that the proliferous inhibitory effect of the TSP on SMMC-7721 human hepatoma cells was augmented with the increased concentrations and prolonged time.2.Annexin V/PI double stainning was used to determine the natural compounds on TSP induced apoptosis. The results showed that production which TSP induced-apoptosis when TSP treated SMMC-7721 human hepatoma cells for 48h.3.The qRT-PCR assays were performed to test the expression of apaf-1, caspase-3, caspase-9, cytochromeC, Bax and p53, which are tumor related genes. The results showed that TSP with the concentration of 25μg/ml,50μg/ml, 100μg/ml,200μg/ml could significantly or extremely significantly promote regulated expression of caspase-3 and cytochromeC mRNA in the proliferation of SMMC-7721 human hepatoma cells; when SMMC-7721 human hepatoma cells was treated with TSP for 72h, TSP could down-regulated expression of apaf-1 and p53 mRNA, and reduced 54.1%,56.25%, respectively; when SMMC-7721 human hepatoma cells was treated with TSP for 72h, and the concentration of 200μg/ml could significantly promote regulated expression of Bax mRNA, and increased 43.9%. And TSP could not significantly promote regulated expression of caspase-9 mRNA.4.The western blot assays were carried out to examine the expression of caspase-3 and cytochromeC proteins. The results found that treatment with 100μg/ml TSP could remarkably increased caspase-3 and cytochromeC protein expression in a concentration-dependent and prolonged time manner, and increased 68.18%,75.47%, respectively.[Conclusion]TSP significantly inhibited the proliferation of SMMC-7721 human hepatoma cells, and up-regulated the expression of tumor-regulated caspase-3, cytochromeC genes and proteins. TSP could down-regulated the expression of apaf-1 and p53 genes. Our data showed that TSP exhibited anti-tumor effect by suppressing or promoting tumor regulated-genes, such as apaf-1, caspase-3, cytochromeC, and p53, suggesting that TSP has certain anti-tumor effect.