Study On Antitumor Activity Of Ginsenoside Combined Transforming Components

ObjectiveThis paper explores the cytotoxic effects of ginsenosides conversion components (sample A) on five tumor cells; On the basis of this study, discusses the anti-proliferation and promote apoptosis and its influence on HepG2 cell cycle that sample A do to the human hepatoma cell line HepG2, and explore the possible mechanism. The study provides a theoretical basis for the clinical and experimental evidence, for the anti-tumor effects’ targets and clinical applications of sample A.MethodsCulture the tumor cells by the conventional methods, select the logarithmic phase cells which grow in good condition to study.①In the experiment, five different concentrations of sample A of Tet 10,20,40,80,160ug/ml, were selected as experimental groups, which were respectively co-cultivated with HepG2 cells and the remaining cells for 24,48,72 hours. Then undergo MTT to find their cellular toxicities.②Based on the experiment ①, choice HepG2 cells as a follow-up research object. Three different concentrations of sample A of Tet 20,40,80ug/ml were selected as experimental groups, which were respectively co-cultivated with HepG2 cells for 48 hours. The cell morphological change of apoptosis was observed under inverted microscope and confocal microscope. The percentage of cell apoptosis, the cycle’s change of DNA were examined by flow eytometry.③Three different concentrations of sample A of Tet 20,40,80ug/ml, were selected as experimental groups, which were respectively co-cultivated with HepG2 cells for 48 hours. Western blot was used to assess the protein expression of Bcl-2 and Bax in HepG2 cells.Results①The studies demonstrate that sample A inhibited proliferation of HepG2 cells, MGC-803 cells, SGC-7901 cells, Hela cells and Caski cells. Among them, the cellular cytotoxicity of sample A on HepG2 cells was strongest. And the IC50 values on HepG2 cells were (98.68±2.82) μg/ml, (42.15 ±1.64) μg/ml, and (34.26 ±4.45) μg/ml. When its concentration was 40μg/ml, it had the strongest effect in 48h.②After HepG2 cells were treated by 20μg/ml,40μg/ml,80μg/ml sample A for 48 hours, morphological changes were observed by inverted microscope. Apoptosis of HepG2 cells was analyzed by Hochest 33342 fluorescent staining. It is thus evident that morphologic characteristics of typical apoptosis in electron microscope. Numerous round cells, shrinkage of cellular membrane and dead cells, karyopyknosis, chromatic agglutination adjacent to the nuclear membrane and apoptotic body were observed.③After adding 20,40 and 80μg/ml sample A in HepG2 cells, use flow cytometry to test the cellular proliferation index. In company with the rising of sample A concentration gradient, the inhabition rate gradually increasedin a dose-dependent manner, the values from blank control group to 20,40 and 80μg/ml group sequentially were 9.14%,20.79% and 31.53%.④After adding 20,40 and 80μg/ml sample A in HepG2 cells, use flow cytometry to test the cell-cycle. Sample A can interfere the cell-cycle phase in HepG2 cells and cause cell remain in G0/G1-phase, and this effect shows dosage-dependent.⑤After HepG2 cells were treated by 20μg/ml,40μg/ml,80ug/ml sample A for 48 hours, the expression of Bcl-2 decreased and that of Bax increased.ConclusionsSample A have anti-proliferative effects on five tumor cells. Sample A can inhibit proliferation, induce apoptosis, and interfere with cell cycle progression in HepG2 cells, and its functions relate to Bcl-2, Bax factor.