Study On The Inhibitory Effect Of Oxymatrine Combined With UTD1 On The Proliferation Of Human Breast Cancer MCF-7 Cell Lin

Objective: This project is designed to study the inhibitory effect of oxidized matrine combined with UTD1 on human breast cancer cell strain MCF-7.Materials and Methods: 1.Retrieve compound components and targets of bitter ginseng by TCM pharmacology database and analysis platform(TCMSP);select breast cancer targets by Gend Cards and OMIM databases,and construct visual network map of drug-key compound-disease target by Cytoscape software;finally,build PPI protein interaction network,GO enrichment analysis and KEGG pathway enrichment analysis by String database and Bioconductor platform.2.Subjects: human breast cancer MCF-7 cell line.4.Experimental method: CCK-8 method was used to detect the effect of inhibitory proliferation of human breast cancer cell line MCF-7,The drug concentrations of oxidized matrine were set to 6.25μg/ml,12.5μg/ml,25μg/ml,50μg/ml,and 100μg/ml,The IC50 value is also calculated;The effect of UTD1 on human breast cancer cell line MCF-7 by CCK-8,The drug concentrations of UTD1 was set to 250 nmol / L,500 nmol / L,1 μmol/L,and 2.5 μmol/L,And calculate the IC50 value;The inhibitory effect of oxidized matrine combined with UTD1 on MCF-7 was determined by CCK-8,Using 6.25 μg/ml of oxidized matrine in combination with UTD1 of250 nmol / L,500 nmol / L,1 μmol/L and 2.5 μmol/L,respectively,The IC50 values were also calculated.The effects of 6.25 μg/ml of matrine oxidation,0.6 μmol/L UTD1 monotherapy(48h IC50)and MCF-7 cycle and apoptosis in human breast cancer cell lines were determined by flow cytometry.The effect of oxidized matrine combined with UTD1 on the protein expression of AKT,caspase-9,caspase-3 and Bcl-2 was determined by Western Blot,and the apoptosis mechanism was defined.Results:1.A total of 22 compounds are associated with breast cancer disease targets,achieving therapeutic effects on breast cancer diseases mainly by regulating AKT1,TP53,JUK,VEGFA,BCL-2,caspase-3 genes,P13 K / AKT,and AGE / RAGE.2.Different concentrations of oxidized matrine and UTD1 showed significant inhibitory effects on the human breast cancer cell line MCF-7 in a concentration-and time-dependent manner.Oxidative matrine IC5024h: 33.41 μg/ml,IC5048h: 20.1 μg/ml,IC5072h: 17.24μg/ml;UTD1IC5024h: 0.8 μmol/L,IC5048h: 0.6 μmol/L,IC5072 h value: 0.5 μmol/L.3.6.25 μg/ml with 250 nmol / L,500 nmol / L,1 μmol/L,and 2.5 μmol/L had a synergistic inhibition of cell proliferation,and the IC50 of UTD1 decreased to: 0.5nmol/L,0.4nmol/L,0.3nmol/L at 24 h,48h and 72 h,respectively.4.After 48 h treatment with 6.25 μg/ml oxidized bitrine,0.6 μmol/L UTD1 monotherapy and two-drug combined human breast cancer cell line MCF-7,the results showed that cells in the single agent group showed G1 phase arrest,and both G1 and G2 / M cells increased in the combination group,but showed a significant increase in G2 / M phase.5.The combination of 6.25 μg/ml,0.6 μmol/L monotherapy and UTD1 in human breast cancer cell line MCF-7 showed different degrees of apoptosis in all three groups,and in the combined group to promote UTD1-induced late apoptosis,with a significant increase in total apoptosis rate compared with control and monotherapy groups.6.The results of Western Blot experiments showed that the content of P-AKT,Bcl-2 and caspase-9 were significantly decreased and the expression of caspase-3 was increased,suggesting that the inhibitory effect of oxidized matrine and UTD1 on breast cancer cells was achieved by regulating the P13 K / AKT pathway.Conclusion:1.The extract can treat breast cancer by multi-route and multi-target method.2.Both matrine and UTD1 monotherapy have inhibitory effects on cell lines.Combined with UTD1 can significantly enhance the inhibitory proliferation of human breast cancer MCF-7cell lines and arrest cells in G2 / M phase.3.The combination of oxidized matrine with UTD1 was able to increase the apoptosis ratio by inhibiting the P13 K / AKT pathway.