Study On The Mechanism And Related Experiments Of Ginseng Active Ingredients On Three Negative Breast Cancer

Objective:Through systematic network pharmacology and bioinformatics methods combined with experimental verification,to explore the pharmacological substance basis and related mechanism of ginseng in triple negative breast cancer(TNBC),and provide theoretical support and clinical application guidance for ginseng in TNBC.Method:1.Active components and target genes of ginseng.The effective components of ginseng were retrieved by TCMSP database,and the target genes of effective components were obtained by UniProt database.The target gene of ginseng was enriched and analyzed to study its biological function process.2.TNBC microarray and differentially expressed genes.The microarray gene chips related to TNBC were retrieved by GEO database and the differential expression was analyzed to obtain the differential expression genes of TNBC.We visualized the protein-protein interaction network of differentially expressed genes,and got the key genes of the network after topological analysis.3.Combination of effective components of ginseng with TNBC target.Through molecular docking technology,the key genes differentially expressed between ginseng and TNBC were scored.Obtain the most effective components of ginseng,and carry out the next step of cell experiment verification.4.The active components of ginseng act on MDA-MB-468 and MDA-MB-231 cell lines.CCK8 method was used to observe the effect of different concentrations of ginseng active ingredients on the proliferation of human breast cancer cell lines MDA-MB-468 and MDA-MB-23 1.Cell migration and invasion experiments were carried out to detect the migration and invasion of MDA-MB-468 and MDA-MB-231 cells.The expression of key genes in breast cancer MDA-MB-468 and MDA-MB-231 cells treated with different concentrations of ginseng active ingredients was detected by real-time fluorescence quantitative PCR.Results:1.Through tcmsp database,15 kinds of effective chemical constituents of ginseng were retrieved,and 172 target genes of effective chemical constituents were predicted by UniProt Enrichment analysis showed that these target genes are involved in cancer pathway,calcium signaling pathway,cAMP signaling pathway,PI3K Akt signaling pathway,transcription disorders in cancer and other TNBC related pathways.2.Three TNBC microarray gene chips GSE45827,GSE38959 and GSE81838 were screened by GEO database,and 196 differentially expressed genes were obtained.According to the topological analysis,MAD2L1?CCNB1?TOP2A?BUB1?CDC6?KIF11?BUB IB and AURKA are 8 key genes in 196 differentially expressed gene networks.Moreover,pathway analysis showed that It was also found that BUB1 and CCNB1 were co-enriched in the cell cycle pathway,which was closely related to the disease process of TNBC.In this pathway BUB1 regulated CCNB1 by CDH1 in cell cycle.Therefore,BUB1 and CCNB1 are three key genes differentially expressed in TNBC.3.Two key genes(BUB1 and CCNB1)of TNBC and 15 active components of ginseng were used for molecular docking simulation,and it was found that the best docking simulation was between two key genes and ginsenoside Rh2(G-Rh2).Because BUB1,CCNB1 and CDH1 have regulatory relationship,the next walking cell experiment verification.4.The results of CCK8 experiment showed that G-Rh2 could inhibit the proliferation of MDA-MB-468 and MDA-MB-231 cells in breast cancer,and it was time and concentration dependent(P<0.05).The migration and invasion experiments showed that G-Rh2 inhibited the migration and invasion of cells significantly(P<0.01).The results of real-time fluorescent quantitative PCR showed that the expression of BUB 1,CCNB1 and CDH1 in MDA-MB-468 and MDA-MB-231 could be inhibited with the increase of G-Rh2 concentration(P<0.01).Conclusion:This study suggests that G-Rh2,as a key active component of ginseng,can inhibit the proliferation and migration of human breast cancer cell lines MDA-MB-468 and MDA-MB-231.Its mechanism is to regulate TNBC by inhibiting the expression of BUB1,CCNB1 and CDH1.This result can provide molecular basis for the further study of ginseng in the treatment of TNBC,and also shows the feasibility of using network pharmacology to connect large-scale target data as the mechanism of traditional Chinese medicine interference.