Study On The Anti-tumor Effect And The Mechanism Of Total Flavones Extracts From Ampelopsis Megalophylla

Ampelopsis megalophylla Diels et.Gilg is a medicinal plant of the genus Ampelopsis,its medicinal part is tender leaves and stems.It is neutral in nature,sour and bitter tasting,with the functions of clearing heat and cooling blood.It is mainly distributed in hubei,shanxi,yunnan and other places in China,used in Enshi Hubei province.In preliminary research,our team has been completed extracted and isolated to obtain total flavones extracts through the percolation method.The six kinds of tumor cells,Hela,A549,MCF-7,HepG2,A2780 and SW620 cells were selected to screen anti-tumor activities for the total flavones extracts from A.Megalophylla via MTT method in vitro.The experimental results showed that the total flavones extracts from A.Megalophylla inhibited the proliferation of six tumor cells in dose-and time-dependent manners.And the more sensitive cell lines were MCF-7,A2780 and SW620 cells,IC50 were 28.47,18.41 and21.85 μg·mL-1 respectively.Moreover,the potential mechanism of cytotoxic effect had been evaluated on MCF-7 cell induction by the total flavones extracts from A.Megalophylla.On the basis of Hochest33258 staining,propidium iodide(PI)staining,Annexin V-FITC/PI double marking,rhodamine123(Rh-123)and western blot analysis,the results indicated that the total flavones extracts from A.Megalophylla induced apoptosis in MCF-7 cells resulting in inhibition,apoptotic bodies could be found under fluorescent microscopy,apoptoticpercent presented a dose-dependent in a certain range.Hochest33258 staining and Annexin V-FITC/PI double marking suggested that the total flavones extracts from A.Megalophylla could induce apoptosis from two aspects of qualitative and quantitative respectively;The total flavones extracts from A.Megalophylla could make MCF-7 cells cycle arrested in G2/M phase by PI staining.Meanwhile,the mitochondrial membrane potential was markedly decreased.Western blot assay showed that the expression of Caspase8,Bax,Cytc,Apaf-1,Caspase9 and Caspase3 proteins dramatically increased after incubation with the total flavones extracts from A.Megalophylla,while the expression of Bcl-2 decreased,then the ratio of the Bcl-2 / Bax declined.Based on the analysis of above results,the mechanism of the total flavones extracts from A.Megalophylla induced apoptosis in MCF-7 cell may be related to the mitochondrial pathway.