Network Pharmacology Analysis And Experimental Verification Of Quercetin Enhancing Radiosensitivity Of Nasopharyngeal Carcinoma

Objectives: The incidence of nasopharyngeal carcinoma(NPC)is high in Guangdong and Guangxi in China.Radiotherapy is the first-line treatment for NPC.xiaochaihu decoction(XCHD)has anti-tumor effect.The aim of this study is to predict the active components,core targets and key signaling pathways of Xiaochaihu decoction in the treatment of nasopharyngeal carcinoma(NPC)using network pharmacology method,and to verify that quercetin can enhance the radiosensitivity of NPC by constructing radioresistant NPC cells in vitro,providing theoretical basis for exploring the clinical treatment of NPC.Methods:(1)The differentially expressed genes of NPC radioresistance were identified by GEO database;Systematic pharmacology of traditional Chinese Medicine(TCMSP)was used to find the active ingredients and targets of Xiaochaihu decoction.Venn diagram was used to obtain intersection genes between effective drug genes and NPC radioresistance differentially expressed genes.Cytoscape software was used to establish the "drug-component-target-disease-network",STRING database and Cytoscape software were used to draw the PPI network,and clinical experiments were conducted to verify the target.R software was used to perform GO and KEGG analysis of intersection genes.Clinical experiments were also conducted to verify the correlation between target genes PPARG(peroxisome proliferator-activated receptor gamma),CASP3(caspase-3)and radiotherapy.(2)Establishment of radioresistance model of nasopharyngeal carcinoma,and detection of biological behavior of normal nasopharyngeal carcinoma CNE-2 cells and nasopharyngeal carcinoma resistant cell CNE-2R by cell scratch test;(3)To verify the accuracy of network pharmacology analysis,CNE-2R cells were treated with different concentrations of quercetin(QC).CCK-8 assay was used to detect the proliferation of CNE-2R cells under the intervention of quercetin,and flow cytometry was used to detect the apoptosis of CNE-2R cells.Results:(1)A total of 2731 differentially expressed genes in radioresistance were obtained from the GEO database,including 1551 up-regulated genes and 1180 downregulated genes.Xiaochaihu Decoction had 246 active ingredients,and 38 intersection genes were obtained by combining the effective drug genes with the differentially expressed genes of NPC.According to the analysis of drug-component-target-disease network,the main active ingredients of Xiaochaihe Decoction were quercetin,kaempferol,hanxanthin,formononetin,isoflavone,stigmasterol,naringenin,β-carotene,and xanthin.PPI analysis showed that the core target genes of Xiaochaihu decoction in improving the radiosensitivity of nasopharyngeal carcinoma were PPARG,RXRA(retinoid X receptor alpha),ACHE(acetylcholinesterase,acetylcholinesterase).ACHE),monoamine oxidase B(MAOB),CASP3,CYP3A4 and BAX target genes,which may enhance the radiosensitivity of nasopharyngeal carcinoma,and the correlation between target genes PPARG and CASP3 and radiotherapy has been verified by clinical practice.(2)The morphology,growth rate and migration rate of CNE-2R cells were changed.(3)The results of CCK-8 assay showed that quercetin inhibited the proliferation of CNE-2R cells in a concentration-and time-dependent manner,and promoted the early apoptosis of CNE-2R cells.Conclusions: The core component of Xiaochaihu Decoction in enhancing the radiosensitivity of nasopharyngeal carcinoma is quercetin,and the key targets are seven target genes,PPARG,RXRA,ACHE,MAOB,CASP3,CYP3A4 and BAX.The target genes PPARG,RXRA and CASP3 were verified by molecular docking.In vitro cell experiments showed that quercetin inhibited the proliferation of nasopharyngeal carcinoma CNE-2R cells and promoted the early apoptosis of CNE-2R cells.