Exploring The Pharmacodynamic Material Basis And Molecular Mechanism Of Xihuang Pill In Breast Cancer Based On Tumor Metabolic Reprogramming

Research purposes:Based on the theory of tumor metabolic reprogramming,this study explored the pharmacodynamic material basis and complex molecular mechanism of Xihuang Pill’s intervention in breast cancer by using a multidisciplinary cross-integration research model such as bioinformatics,integrative pharmacology and pharmacological experiments.It is hoped that the changes in the metabolic environment of breast cancer will be explained in depth,and the interaction mechanism of the breast metabolic network will be visualized,so as to provide a theoretical basis for the modern scientific research and clinical application of Xihuang Pills,and to provide ideas for the construction of a modern molecular mechanism research model for Chinese medicine to interfere with malignant tumors.Research method:1.To explore the "traditional Chinese medicine-component-target-disease" characteristics of Xihuang Pill under the platform of integrated pharmacology.First,the active components of each Chinese medicine in Xihuang Pills were comprehensively sorted out by searching the Chinese medicine research platforms of "TCMSP","TCMID","TCM@taiwan" and literature databases of "Pubmed" and "National Knowledge Infrastructure".Then,based on platforms such as "TCMSP" and "Swiss Target Prediction",the putative therapeutic targets of the active components of Xihuang Pills were predicted,The biological characteristics and pathway characteristics of the target were explained by "KEGG" and "GO" analysis;finally,interaction networks such as "TCM-component","component-target","target-pathway" and " target-target " were constructed.Analyze network characteristics through topology analysis and module analysis.2.Bioinformatics technology to explore the metabolism-related molecular pathogenesis of breast cancer.First,the breast cancer data was downloaded from the "TCGA" database,and statistical methods were used to screen and organize the data to obtain the relevant differentially expressed genes of breast cancer;then,from the The GSEA database downloaded the genes related to metabolic pathways,obtained the differentially expressed genes of breast cancer metabolism after taking the intersection,and analyzed and sorted the genes.Finally,based on the "R language" analysis,the genes related to survival in breast cancer and triple negative breast cancer were obtained.3.Screening the metabolism-related genes that Xihuang Pill acts on breast cancer based on computer molecular docking technology.First,the protein energy crystal of the gene was downloaded from the RCSB PDB platform,and the "mol2" format of the active ingredient of Xihuang Pill was downloaded from the TCMSP platform.Then,the "drug-target" was molecularly docked using the Surflex-Dock module of the SYBYL X.2.1.1 software.,and obtained the Surflex-Dock score;finally,the active ingredients and genes of Xihuang Pills with the highest binding degree were obtained,and the docking results were further optimized using Py MOL software.4.Experiment to verify the drug activity of Xihuang Pill on breast cancer cells.First,the CCK8 proliferation assay was used to explore the effect of different concentrations of Xihuang Pill on the proliferation of breast cancer cells MCF-7 and MDA-MB-231 cells;then,scratch assay and invasion assay were used to detect the effect of different concentrations of Xihuang Pill on MCF-7 And MDA-MB-231 cell migration ability and invasion ability;Finally,q RT-PCR and western blot experiments were used to detect the expression of metabolism-related genes andproteins in breast cancer cells under the intervention of Xihuang Pill.Research result:1.By integrating pharmacology methods such as database search,literature mining and protein network construction,a total of 53 small molecules of drug activity of Xihuang Pill were obtained,and 318 therapeutic targets of Xihuang Pill were presumed and predicted.These targets are mainly enriched in cancer and metabolism-related pathways,including PI3K-Akt signaling pathway,proteoglycan in cancer,MAPK signaling pathway,micro RNA and other pathways in cancer,AGE-RAGE signaling pathway,lipid and atherosclerosis pathway,etc.These studies It shows that Xihuang Pill can be used for clinical treatment of tumors,metabolic diseases and other diseases.2.By analyzing breast cancer high-throughput big data,445 differentially expressed genes related to breast cancer metabolism were obtained,including 226 up-regulated genes and 219 down-regulated genes,of which 35 genes were associated with prognosis.445 genes are involved in the production of various metabolites and enzymes related to metabolic processes,and the main metabolic pathways involved include purine metabolism,pyrimidine metabolism,carbon metabolism,amino sugar and nucleotide sugar metabolism,arginine and proline Metabolism,fructose and mannose metabolism,glycolysis/gluconeogenesis,cysteine and methionine metabolism and other pathways indicate that in the molecular mechanism of the occurrence and development of breast cancer,the metabolic network has been remodeled,and the importance of the metabolic network of breast cancer has been remodeled.Programmed interventions may provide insights into the clinical treatment of breast cancer.3.Molecular docking results show that multiple active components in Xihuang Pill can act on the same target,and the same target can be regulated by multiple components.For example,the docking results of 10 active small molecules in Xihuang Pill and GGCT are greater than 7 points,and there are The docking results of 6 active small molecules of Xihuang Pill with NUDT5 are greater than 7,which explains the "multi-component-multi-target" characteristics of traditional Chinese medicine compound intervention in diseases,indicating that Xihuang Pill can interfere with tumor metabolic reprogramming,thereby inhibiting the progression of breast cancer.4.The experimental results show that Xihuang Pill can inhibit the proliferation,migration and invasion of breast cancer cells.Subsequently,q RT-PCR and Western Blot experiments were used to verify the molecular mechanism of Xihuang Pill’s intervention on the metabolic genes of the classical metabolic pathway.After the intervention of Xihuang Pill,the expression of GGCT and NUDT5 in MCF-7 cells was inhibited,and the expression of ALDOC and SHMT1 in MDA-MB-231 cells was activated.This study found that GGCT and NUDT5 were highly expressed in breast cancer,and were associated with poor prognosis.related.After the intervention of Xihuang Pill,the expression of these genes was altered,and the tumor metabolic reprogramming that promoted breast cancer was turned into a favorable factor for inhibiting breast cancer progression.Furthermore,we applied this integrative pharmacology research model to triple-negative breast cancer and found that ALDOC and SHMT1 were lowly expressed in triple-negative breast cancer and correlated with poor prognosis.It further proved the feasibility and effectiveness of this research model in exploring the molecular mechanism of TCM anti-tumor.In conclusion:Based on the integrated pharmacology model,this study explained the pharmacological properties and clinical application of Xihuang Pills from four aspects: traditional Chinese medicine,components,targets,pathways,and diseases;Based on high-throughput big data analysis of bioinformatics,taking tumor metabolic reprogramming as an entry point,the molecular mechanism of tumor metabolic reprogramming in breast cancer was explained;Based on molecular docking and experimental verification technology,it is shown that the intervention of Xihuang Pill on metabolic reprogramming of breast cancer involves a variety of metabolic processes,including glutathione metabolism(GGCT),purine metabolism(NUDT5),folic acid metabolism(SHMT1),Glycolysis(ALDOC),etc.,thereby promoting the transformation of the metabolic network of breast cancer to a favorable factor that inhibits the progression of breast cancer,thereby inhibiting the molecular mechanism of the occurrence and development of breast cancer.And the multidisciplinary research mode adopted in this study provides ideas for the modern research of traditional Chinese medicine anti-tumor.