Four Gentlemen Decoction Combined With 5-fu Induced Experimental Study Of Human Gastric Cancer Cell Apoptosis And Its Mechanism

Objective: To investigate the apoptosis and its mechanism of human gastric carcinoma cell line SGC-7901 induced by SiJunZi decoction (SJZD) and 5-FU. METHODS: The Chinese Herbs' Serum Pharmacological method and cellculture in vitro were used in this study.Human gastric carcinoma cells SGC-7901 were treated by the blood serum containing SJZD and 5-FU for 24 hours.Electron microscopy were used for observing the morphological change of the cells.The rate of inhibition of gastric cancer cell growth was tested by the method of tetrazolium (MTT).The cycle of growth and the index of apoptosis of gastric carcinoma cell were detected by flow cytometry. The immunohistochemical staining method was used for examining the expression of p53,Bcl-2 and Bax. The content of the blood serum containing SJZD were determinated by High pressure liquid chromatography.RESULTS: The serum containing SJZD and 5-FU used together could inhibitthe proliferation of human gastric carcinoma cells SGC-7901.The rates of inhibition increased with dosage dependence.(After 24 hours of exposure to 10 percent of the serum containing SJZD of three different dosage and 5-FU.)SGC-7901 exhibitted morphological features of apoptosis,including cell shrinkage,grew against the wall and the peeled off gradually,which showed the characters of cells apoptosis like chromatin condensation.All these were more significantly than control group.By flow cytometry (FCM) analysis ,the apoptosis index (AI) of groups of two drugs used together were higher than them used single and have a significant difference compared with that of control group. Most cells were arrested in G1/S phase by 5-FU used single and arrested in S phase by the serum containing SJZD of high dosage and 5-FU used together group.The expression of Bcl-2 was down regulated,the expression of Bax was up regulated,and the expression of p53 did not have a significant change with a comparison with control group.Through the method of High pressure liquid chromatography, Ginsenoside Rg1 were determinated in the serum containing SJZD of high dosage group.CONCLUSION: SJZD and 5-FU used together can induce the apoptosis ofSGC-7901 cells cultured in vitro,and its mechanism might be related to the down-regulation of exprseeion of Bcl-2 and the up-regulation of the expression of Bax,but not to the expression of p53.The anti-cancer effective content maybe was Ginsenoside Rg1.This study will provide the treatment of SJZD combined with western medicine with a theoretic support.