The Screening Of Chinese Medicine Monomers Against Nasopharyngeal Carcinoma And The Study On Effect And Mechanism Of Sanguinarine Against Nasopharyngeal Carcinoma

Purpose: Nasopharyngeal carcinoma(NPC)is a malignant tumor characterized by regional specificity.In this study,we have completed the screening of 70 Chinese medicine monomers.Finally,sanguinarine,a traditional Chinese medicine monomer that effectively inhibits the proliferation of nasopharyngeal carcinoma cells,was selected as the research object.Analyze the feasibility of sanguinarine against nasopharyngeal carcinoma through network pharmacology and conduct research on the biological behavior and mechanism of anti-nasopharyngeal carcinoma.This provides a preliminary theoretical basis for sanguinarine to become an effective anti-nasopharyngeal cancer drug and its subsequent development and utilization.Methods:1.The effect of seventy kinds of traditional Chinese medicine monomers on the vitality and proliferation of nasopharyngeal carcinoma cells(CNE1,CNE2,5-8F and 6-10B)through high-throughput screening of Cell Titerglo method.2.In this study,the method of network pharmacology was used to predict the potential drug target of sanguinarine for nasopharyngeal carcinoma.Then,it was verified by molecular docking.3.The Cell Counting Kit-8(CCK-8)experiment and the cell clone formation experiment were used to determine the effect of sanguinarine on the viability and proliferation of nasopharyngeal carcinoma cells;The cell scratch test,Transwell migration and invasion test were used to detect sanguinarine effects the influence of nasopharyngeal carcinoma cell migration and invasion;Annexin V-FITC/PI fluorescence staining flow cytometry to analyze the effect of sanguinarine on the apoptosis of nasopharyngeal carcinoma cells;Western-blot method to study the cell signal transduction pathway.Results:1.Through the screening of seventy kinds of Chinese medicine monomers,we found that six kinds of Chinese medicine monomers have a good inhibitory effect on the proliferation of nasopharyngeal carcinoma cells.Based on the analysis of the experimental results and the comprehensive evaluation of the research progress and efficacy of the anti-tumor effects of these Chinese medicine monomers by consulting the literature,finally,we choose sanguinarine,which is currently less researched in the field of anti-tumor,as the anti-nasopharyngeal cancer research Object,conduct further molecular research.2.Through the prediction of network pharmacology,a total of thirty-eight potential common targets of nasopharyngeal carcinoma and sanguinarine have been identified.GO functional enrichment analysis showed that protein serine/threonine kinase activity,histone kinase activity,integrin binding,protein tyrosine kinase activity and cell adhesion molecule binding ranked first.KEGG enrichment analysis shows that the potential pathway is mainly involved in the PI3K-Akt signaling pathway.The"drug-target-disease-pathway"network model diagram points out key genes such as MAPK10,MAPK14,JAK2,BRAF,GSK3B,MET,HSP90AA1,and SRC.According to the results of molecular docking,sanguinarine has a strong binding ability with MAPK10 and MAPK14.3.Sanguinarine inhibits the proliferation of nasopharyngeal carcinoma cells(poorly differentiated CNE2 cells and highly metastatic 5-8F cells)in a time and dose-dependent manner.Cell clone formation experiments showed that sanguinarine reduced the cloning ability of nasopharyngeal carcinoma cells.In addition,sanguinarine has a certain inhibitory effect on the invasion and migration of nasopharyngeal carcinoma cells.Annexin V-FITC/PI fluorescence staining flow cytometry experiments showed that sanguinarine promoted the apoptosis of nasopharyngeal carcinoma.Western blotting experiments showed that the mechanism of sanguinarine against nasopharyngeal carcinoma is mainly related to the inhibition of mTOR signal transduction.Conclusion:1.Through the screening of seventy kinds of Chinese medicine monomers,it is found that alkaloids,flavonoids,steroids,terpenes(sesquiterpenes,monoterpenes,diterpenes,triterpenes)Chinese medicine monomers have a strong inhibitory effect on cell proliferation.2.In addition,we found that sanguinarine may become a new high-efficiency anti-nasopharyngeal cancer monomer of traditional Chinese medicine,which has certain reference value for our follow-up in vivo research and clinical trials.