| In recent years, people are looking for new natural medicines inhibiting tumor occurrence and metastasis. Saponins are widely distributed in the plant kingdom, and have good antitumor effect. Saponins could not only inhibit the proliferation of cancer, but also enhance immunity, and inhibit tumor metastasis. Gynostemma Pentaphyllum (GP) belongs to the medicine food homology plant, and gypenosides are the main active ingredient. In the aspect of anti-cancer, there have been many studies about inhibition of tumor cell of the total saponins extracted from GP at present, however, the monomer gypenosides are still rarely reported and their underlying molecular mechanisms have not been well defined. This subject studied the mechanism of induction cancer cells apoptosis, inhibiting metastasis of three monomer gypenosides N-2, J-0-1, J-D-40 by applying different assays.In this study, inhibition of human hepatocarcinoma HepG2 cells proliferation was determined by MTT assay; Scanning electronic microscopy (SEM) to observe changes in cell morphology; nuclear morphology by DAPI staining was observed with fluorescence microscope; Apoptosis was assessed by flow cytometry after staining the cells with annexin V-FITC; Mitochondrial transmembrane potential was detected by Rhodamine-123 staining; Experiments of wound healing and Transwell were used to detect the ability of cell migration, cell invasion.The expression of p53, MMP-2, MMP-9, and HIF-la mRNA by RT-PCR and proteins by Western blotting were also performed.The results show that gypenosides N-2, J-0-1, J-D-40 have the ability to induce cell apoptosis and inhibit metastasis. Samples inhibited HepG2 cells proliferation, and the inhibition was dose-dependent; After treatment with ginsenoside Rg3 200μmol/L, N-2 200 μmol/L, J-0-1 100 μmol/L, J-D-40 50 μmol/L, the HepG2 cells occured typical apoptosis morphology. Samples increased the number of apoptosis cells, elevated the mitochondrial membrane potential. The ability of cell adhesion, cell migration rate, cell invasion were also reduced. The expression of gene p53, MMP-2, MMP-9 and HIF-la mRNA levels were reduced significantly, the same is true for protein levels. Therefore gypenosides N-2, J-0-1, J-D-40 can down-regulation p53, MMP-2, MMP-9 and HIF-la to achieve the purpose of inhibiting cell proliferation, metastasis and invasion of HepG2 cells. |
