Effect And Mechanism Of Xiaotan Sanjie Decoction In The Treatment Of Gastric Cancer

Research objective:The high morbidity,high mortality,poor prognosis and drug resistance of gastric cancer pose a severe test for routine surgical treatment and chemotherapy.According to the theory of traditional Chinese medicine,gastric cancer generally belongs to deficiency of origin and excess of sign,which means empty of spleen and retention of phlegm.Traditional Chinese medicine has the advantages of "supporting the right and expelling the evil" and "consolidating the foundation and clearing the source".Xiaotan Sanjie decoction is an effective traditional Chinese medicine prescription developed on the basis of the ’phlegm theory’ of gastric cancer.Previous studies have shown that traditional Chinese medicine prescriptions have multi-target and multi-pathway anticancer effects.Therefore,this study is based on the method of network pharmacology to predict the molecular mechanism of Xiaotan Sanjie Decoction in the treatment of gastric cancer,and combined with the experiment of the effect of Xiaotan Sanjie decoction containing serum on gastric cancer cell SGC-7901,in order to explore the effect and mechanism of Xiaotan Sanjie Decoction in the treatment of gastric cancer,and provide a theoretical basis for subsequent experimental research.Methods:1.Network pharmacology predictionWe collected and screened the active components of Xiaotan Sanjie Decoction through Traditional Chinese Medicine Systems Pharmacology database and analysis platform(TCMSP)to predict the target of the components through TCMPS database;we screened gastric cancer targets in NCBI gene database(NCBI),Gene Cards database and OMIM database(Online Mendelian inheritance in Man,OMIM);we took drug targets and disease targets to Make Venn diagram,and used common targets to make PPI network diagram in String database and cytoscape3.8.0,and screened core targets and core genes by MCODE cluster analysis;we imported the relevant results into Cytoscape3.8.0 to draw the component-disease-target network diagram;the common targets of drug and diseases,we used Omicshare and Metascape databases to do the GO analysis and KEGG analysis.2.Effect and mechanism of Xiaotan Sanjie Decoction on gastric cancer SGC-7901cells2.1 Rats were perfused with Xiaotan Sanjie Decoction to prepare drug containing serum for subsequent cell experiments.2.2 SGC-7901 cells were treated with serum containing different concentrations of Xiaotan Sanjie decoction.The effect of serum containing different concentrations of drugs on SGC-7901 cell proliferation was detected by CCK-8 method,and the appropriate concentration and action time of serum containing drugs were screened.2.3 SGC-7901 cells were treated with 4% and 8% Xiaotan Sanjie decoction containing serum for 72 hours,and their effects on cell cycle and apoptosis were detected by flow cytometry.2.4 Western blot assay was used to detect the effect of Xiaotan Sanjie decoction containing serum at 4% and 8% concentrations on the expression of apoptosis related factors Bcl-2,Bax,caspase-3 and caspase-9 in SGC-7901 cells,and to explore the possible mechanism of Xiaotan Sanjie Decoction inducing apoptosis.Results:1.The "component-disease-target" network of Xiaotan Sanjie Decoction in the treatment of gastric cancer involves 216 common targets.Through PPI network construction and MCODE cluster analysis,9 gene clusters were obtained and 10 core genes were screened,namely STAT3,VEGFA,AKT1,Jun,CASP3,MMP9,IL6,TNF,PTGS2 and IL1 B.2.The function enrichment of GO suggests that the biological process and function of Xiaotan Sanjie Decoction in the treatment of gastric cancer focus on cell process,biological regulation,oxidative stress response,biological metabolism,multicellular biological process and so on.3.KEGG pathway enrichment analysis indicates that Xiaotan Sanjie Decoction mainly involves cancer-related pathways,IL-17,mitogen activated protein kinase(MAPK),phosphatidylinositol 3-kinase / protein kinase B(PI3K-Akt),p53,tumor necrosis factor(TNF),apoptosis,cell cycle and other signal pathways in the treatment of gastric cancer.4.The results of CCK-8 method showed that Xiaotan Sanjie decoction could effectively suppress the proliferation of gastric cancer SGC-7901 cells in a time and concentration dependent manner.5.The results of flow cytometry showed that with the increase of the dose of Xiaotan Sanjie decoction,its effect of blocking SCG-7901 cell cycle and inducing apoptosis gradually increased.6.The Western blot test showed that after a 72-hour intervention treatment of Xiaotan Sanjie Decoction medicated serum on SCG-7901 cells,the expression of apoptosis suppressing related protein BCL-2 decreased in a dose-dependent manner,while the expression of apoptosis promoting related proteins Bax,Caspase-3 and caspase-9 increased in a dose-dependent manner.Conclusion:1.The "component-disease-target" network of Xiaotan Sanjie Decoction in the treatment of gastric cancer involves 216 common targets.Through PPI network construction and MCODE cluster analysis,9 gene clusters were obtained and 10 core genes were screened,namely STAT3,VEGFA,AKT1,Jun,CASP3,MMP9,IL6,TNF,PTGS2 and IL1 B.2.Xiaotan Sanjie decoction has multi-functional,multi-target and multi-channel effects on gastric cancer.KEGG pathway enrichment analysis indicates that Xiaotan Sanjie Decoction mainly involves cancer-related pathways,IL-17,MAPK,PI3K-Akt,p53,TNF,apoptosis,cell cycle and other signal pathways in the treatment of gastric cancer.3.The serum containing Xiaotan Sanjie decoction can effectively inhibit the proliferation of gastric cancer SGC-7901 cells,block the process of cell cycle and inducing apoptosis.The mechanism may be to down regulate the expression of apoptosis related protein BCL-2 and up regulate the expression of apoptosis promoting related proteins Bax,Caspase-3 and caspase-9.4.The target and molecular mechanism of Xiaotan Sanjie Decoction in the treatment of gastric cancer may be related to the biological process and signal pathway of regulating cell proliferation,apoptosis and cell cycle.