The Mechanism Of Glycitein Induced Human Gastric Cancer AGS Cells Apoptosis

Gastric cancer is a common malignant tumor and seriously endangers human health.Every year,about 1 million people were diagnosed all over the world.At present,680000 people were diagnosed in China.Glycitein is an important nutrient component in soybean and its products.Glycitein has a variety of biological activities and has attracted the attention of many researchers.But its anticancer mechanism and effective concentration are not clear.Therefore,the objection of this study is explain anti-cancer effects of glycitein on gastric cancer cell and potential molecular mechanisms through the toxicology,molecular biology and cell biology method.It provides new ideas for the development of functional factors and the development of new anti-tumor functional food.In this study,cell counting kit-8 assay was used to detect the toxic effect of glycitein on kinds of human gastric cancer cells and its toxic effect on kinds of normal human cells.The apoptosis of human gastric cancer cells was detected by Hochest 33342/PI double staining,JC-1 mitochondrial membrane potential fluorescence staining,Annexin v-fitc/PI double staining and flow cytometry.The effects of glycitein on the relative content of DNA by flow cytometry and the expression of cycline-related proteins were explored by western blot in AGS cells.The effect of glycitein on expression of apoptosis-related proteins in human gastric cancer cells was determined by western blot.Western blot was used to detect the expression levels of signaling pathway proteins p38,JNK,ERK,NF-?B and STAT3.The regulation of MAPK signaling pathway,NF-?B signaling pathway and STAT3 signaling pathway after treatment of human gastric cancer cells with glycitein and pathway protein inhibitors was detected by western blot.DCFH-DA,flow cytometry and western blot were used to detect the level of ROS in AGS cells.Cell counting kit-8 assay results showed that glycitein decreased the survival rate of kinds of human gastric cancer cells in a different concentration and dose-dependent manner,and the difference was statistically significant compared with the positive control 5-FU.Hochest 33342/PI double staining and Annexin v-fitc/PI double staining result showed that apoptotic morphology of AGS cells changed significantly and the apoptotic proportion increased after the treatment with glycitein.In addition,glycitein had no significant toxic on all four types of normal human cells,and significantly lower than control group.Glycitein reduced mitochondrial membrane potential,inhibiting the expression of anti-apoptotic protein Bcl-2 and up-regulating the expression of pro-apoptotic protein Bax,activated caspase-3 and PARP,and finally caused the apoptosis of AGS cells.At the same time,flow cytometry and western bolt results showed that glycitein caused G0/G1 phase cells arrest in human gastric cancer AGS cells by decreasing the expression level of cyclinD1,cyclinE and CDK2/4/6 proteins,and increasing the expression level of P21 and P27.Glycitein up-regulated the protein expression levels of p-p38 and p-JNK in human gastric cancer AGS cells,and inhibited ERK,NF-?B and STAT3 signaling pathway.Glycitein promoted the production of reactive oxygen species,caused cell apoptosis in human gastric cancer AGS cells.These findings suggested that glycitein induced AGS cell apoptosis and G0/G1 phase cell cycle arrest via ROS-related MAPK/STAT3/NF-?B signaling pathways.Thus,glycitein has the potential to a novel targeted therapeutic agent for human gastric cancer and theoretical basis for the development of functional factors the development of new anti-tumor functional food.