Study On The Mechanism Of Taohong Siwu Decoction In The Treatment Of Breast Cancer Based On Network Pharmacology

ObjectiveThe incidence rate and mortality rate of breast cancer(BC)are the first and fifth of the female malignant tumors in China respectively.They both show a rising trend.They have become a serious disease threatening women’s health in China and even the world.Tao-Hong-Si-Wu Decoction consists of six herbs:Prunus persica(L.)Batsch(Taoren,TR),Carthamustinctorius L.(Honghua,HH),Angelica sinensis(Oliv.)Diels(Danggui,DG),Ligusticum chuanxiong hort(Chuanxiong,CX),Paeoniae Radix Alba(Baishao,BS),and Rehmanniaglutinosa(Gaertn.)DC.(Shudi,SD).Clinical often on the basis of this prescription and combined with drugs to treat breast cancer.However,there is no study on the material basis and mechanism of its therapeutic effect.Therefore,this study uses the network Pharmacology Method to explore the main material basis of THSWD in the treatment of breast cancer,and then uses in vitro cell experiments to verify and elaborate the main targets and pathways of its therapeutic effect,and to provide the thinking and the way of clinical treatment for breast cancer reference resources.Methods1 Analysis of target and pathway of THSWD in the treatment of breast cancer based on network pharmacologyStudy on the network pharmacology of THSWD based on the 107 components of THSWD identified by UPLC-Q-TOF-MS~E.The active components of THSWD were screened according to the principle of similar drugs.Using Pharm Mapper database to predict the target of serum prototype components,using Gene Cards and OMIM database to predict the target of breast cancer,These common targets are the potential target of THSWD acting on breast cancer.The key components of THSWD in the treatment of breast cancer were determined by using the software of Cytoscape to visualize the results.Using the STRING online platform to construct the protein-protein interaction(PPI),the key target genes were selected.The GO analysis and KEGG pathway analysis of the common targets were carried out by using David database.Auto Dock vina software was used for molecular docking verification of the selected key components and key targets.2 Intervention effect of THSWD on breast cancer cellsThe serum containing THSWD was prepared,and the blank serum was set as the negative control group.CCK8 method was used to detect the effect of the serum containing THSWD(2.5%,5%,10%,20%,30%,40%,50%)on the proliferation of MCF-7 and MDA-MB-231 cells at 24,48,72 hours,to find out the best dosage and the best intervention time.The apoptosis of MCF-7 and MDA-MB-231 cells was detected by AO/EB staining and flow cytometry.Transwell migration test was used to observe the effect of THSWD on cell migration.Explore the molecular mechanism of THSWD by RT-qPCR and WB assay.Results1 Analysis of target and pathway of THSWD in the treatment of breast cancer based on network pharmacology27 active components were obtained from 107 in vitro components,Pharm Mapper database obtained 461 component targets,Gene Cards and OMIM database obtained3264 breast cancer related targets,218 targets related to the treatment of breast cancer with THSWD were obtained.12 key components of THSWD in the treatment of breast cancer were obtained by using Cytoscape software to construct the“component-target”network.Using the STRING online platform to construct the protein-protein interaction(PPI),the 14 key target genes were selected.The GO analysis and KEGG pathway analysis of the common targets were carried out by using David database.The results show that THSWD in the treatment of breast cancer by affecting Ras,FoxO,PI3K-Akt and other signaling pathways.The results of molecular docking showed that the key components and corresponding key target proteins had good binding activity,which increased the reliability of pathway prediction.2 Intervention effect of THSWD on breast cancer cellsThe results of CCK-8 showed that in MCF-7 cells,the optimal concentration of serum containing THSWD was 21.7%,and the optimal action time was 48 h;in MDA-MB-231 cells,the optimal concentration of serum containing THSWD was 19.5%,and the optimal action time was 48 h.The serum containing THSWD and the inhibitor group could promote the apoptosis or necrosis of MCF-7 and MDA-MB-231 cells,and inhibit their migration.Compared with cell control group and normal serum group,the m RNA and protein expression levels of HRAS,mapk1,AKT1,Grb2 and Mapk14 in THSWD group and inhibitor group were decreased.Conclusion1 The results of network pharmacology showed that THSWD could intervene breast cancer by acting on multiple targets and pathways.Luteolin,kaempferol,Senkyunolide E and other 12 compounds may be the core active components of THSWD in the treatment of breast cancer;2 It is revealed that the mechanism of THSWD in the treatment of breast cancer may be related to Ras,FoxO,PI3K-Akt and other signal pathways associated with the core targets HRAS,MAPK1,AKT1,GRB2 and MAPK14.