Study On The Potential Target And Molecular Mechanism Of The Shufeng Jiedu Capsule On Lung Adenocarcinoma Based On Network Pharmacology And Basic Experimental Methods

Background: Lung cancer is the main cause of cancer-related mortality.Non-small cell lung cancer is the main types of lung cancer,accounting for 80-85% of all lung cancer cases.In recent years,with the popularization of lung cancer screening,the incidence of lung adenocarcinoma characterized by peripheral nodules has increased,becoming a new hotspot for the diagnosis and treatment of lung cancer.At present,the standard treatment of lung cancer(including surgery,chemoradiotherapy,targeted therapy and immunotherapy,etc.)has greatly improved the survival rate of patients,but many patients still have poor therapeutic effect and overall survival rate.Traditional Chinese medicine(TCM)has unique advantages in treating tumors.In recent years,more and more studies have begun to explore the potential efficacy and mechanism of TCM in counteracting tumors,especially in finding TCM in counteracting lung cancer.Shufeng Jiedu Capsule is a traditional Chinese medicine used to treat a variety of lung diseases and has also played a great role in the treatment of new coronavirus.At present,domestic and foreign research has not paid attention to the role of this traditional Chinese medicine in cancer treatment.In this study,we try to use biological information technology combined with basic experimental methods to explore the potential target and mechanism of Shufeng Jiedu Capsule against lung adenocarcinoma.Methods: The bioactive components and corresponding target gene information of Shufeng Jiedu Capsules were retrieved from the online pharmacology of traditional Chinese medicine platform(TCMSP)and the gene datasets related to lung adenocarcinoma and normal lung tissues were downloaded from the GEO database and analyzed to obtain differential genes related to lung adenocarcinoma.Based on the results of target gene information and lung adenocarcinoma gene data of traditional Chinese medicine,the potential targets of Shufeng Jiedu Capsule against lung adenocarcinoma were obtained by network pharmacological analysis method and the Shufeng Jiedu Capsule-lung adenocarcinoma-target compound network was constructed.Afterwards,string online database was used to construct protein-protein interaction(PPI)network,and GO and KEGG enrichment analysis of target genes were performed to obtain molecular biological processes related to Shufeng Jiedu Capsule against lung adenocarcinoma and pathway information involved in this process.Shufeng Jiedu Capsule solution was prepared to culture human lung adenocarcinoma A549 cells,and the target and pathway mechanism of Shufeng Jiedu Capsule in the treatment of lung adenocarcinoma were verified in vitro,including CCK-8 to verify the effect on cell proliferation,scratch assay and plate cloning assay to verify the effect on cell migration and differentiation,and AO/PI double staining method to identify the effect on apoptosis and necrosis.Western blot was used to verify the expression of related proteins and involved pathways and specific pathway inhibitors were selected as controls to further investigate the effect of traditional Chinese medicine on a certain pathway.Results: A total of eight drugs were selected from the TCMSP database,which were Patrinia,Isatis indigotica,Bupleurum chinense,Glycyrrhiza uralensis,Polygonum cuspidatum,Lianqiao,Reed root,and Verbena officinalis.In addition,242 gene targets corresponding to eight active ingredients were selected.2438 genes were obtained by analyzing the gene data of lung adenocarcinoma and normal lung tissues in GEO database.955 differential genes were analyzed by R language,of which 682 genes were highly expressed related genes and 273 genes were lowly related genes.The target genes of TCM and disease-related target genes were intersected,and a total of 125 intersection genes were obtained as potential target genes and Shufeng Jiedu Capsule – lung adenocarcinoma – target compound network diagram and protein interaction network diagram were constructed.It is concluded that AKT1,ALB,EGFR,IL6 and HSP90AA1 are the core targets of Shufeng Jiedu Capsule acting on lung adenocarcinoma.A total of187 pathways directly related to cancer development were obtained from KEGG pathway enrichment analysis,with the PI3K/AKT pathway having the highest correlation.GO enrichment analysis suggests that Shufeng Jiedu Capsule is mainly involved in protein kinase-related molecular biological processes.The results also showed that Shufeng Jiedu Capsule could inhibit the proliferation and migration of human lung adenocarcinoma cells,and play a role in the apoptosis of lung adenocarcinoma cells.Western blot showed that Shufeng Jiaotou Capsule could significantly reduce the expression levels of PI3K/AKT pathway-related proteins,and this phenomenon was more significant when PI3K/AKT pathway inhibitors synergized.Conclusion: This study preliminarily confirmed that Shufeng Jiedu Capsule has anti-lung adenocarcinoma ability,Shufeng Jiedu Capsule can inhibit the proliferation and migration of human lung adenocarcinoma cells,and active in lung adenocarcinoma cell apoptosis,which may complete the anti-lung adenocarcinoma effect by inhibiting PI3K/AKT signaling pathway.