Prediction And Experimental Verification Of Resveratrol’s Anti-lung Adenocarcinoma Activity Mechanism

Lung adenocarcinoma is the most common type of lung cancer,accounting for about 40% of all lung cancer cases.Most patients with lung cancer have not been found and diagnosed until the advanced stage,although researchers have made achievements in the pathogenesis and treatment methods of lung adenocarcinoma with the continuous advancement of medical standards.Now,lung adenocarcinoma is still one of the most aggressive and fatal malignancies,and the overall survival rate is less than 5 years.Therefore,finding specific biomarkers related to lung adenocarcinoma has become the key to today’s cancer treatment.Microarrays and bioinformatics can explore the mechanism of cancer action at the genome-wide level.In recent years,with the improvement of genome sequencing technology,the popularization of bioinformatics,and the open use of biological big data,more and more researchers have used bioinformatics to analyze the differential expression of cancer and lung cancer tissues.Early diagnosis and treatment of cancer are of great significance.In this paper,the anti-tumor candidate drug resveratrol is screened by predicting the action mechanism of lung adenocarcinoma activity,and then the in vitro cell experiment is performed to verify the action mechanism of resveratrol against lung adenocarcinoma.Whether resveratrol can be used in the treatment of lung adenocarcinoma can provide a theoretical basis for reference.The research of this paper is mainly divided into two parts:Part 1: Screening of anti-adenocarcinoma drugs based on gene expression profilesObjective:Lung adenocarcinoma is the most common type of lung cancer.This paper analyzes gene expression profile data of lung adenocarcinoma by gene chip and sequencing data to obtain differential genes.Through the analysis of differential genes,the metabolic pathways related to lung adenocarcinoma,the differential expression of lung adenocarcinoma and lung cancer tissues,the prognostic relationship,and the small drug molecules that are most negatively related to the differential genes of lung adenocarcinoma were screened to find potential lung Drug molecules related to carcinogenesis,development or treatment.Method:1.Screening of differentially expressed genes: The data of the four LUCD gene expression series(GSE18842、GSE27262、GSE74706、GSE101929)from GEO datasets were analyzed by GEO2R(a software of GEO datasets,based on open source R statistical programming language).The cutoff value criteria are as follow: adj.P.value< 0.05,| log2 Fold Change | >1.2.Pathway expression analysis: The DAVID database and the KOBAS database were used for functional annotation and signal pathway analysis of the differential genes,and the potential biological pathways related to the development of lung adenocarcinoma were screened.3.CMAP analysis: Compare the obtained lung adenocarcinoma differential genes with the CMAP database,retrieve the small molecular compounds with the most significant negative correlation with lung adenocarcinoma,and analyze their potential therapeutic potential in the treatment of lung adenocarcinoma.4.Lung adenocarcinoma gene expression and its relationship with prognosis:Differential genes were introduced into STRING to construct a protein-protein interaction network map and imported into Cytoscape software.The most representative seven were selected using the local analysis method in the Cytohubba plug-in.Genes MAD2L1,ASPM,NUSAP1,BIRC5,DLGAP5,BUB1,ZWINT.The Oncomine online database was used to analyze gene expression,and the online data tool GEPIA was used to verify the expression of the seven genes in the TCGA database in lung adenocarcinoma tissues and normal tissues,and to analyze their relationship with prognostic survival.Conclusion: A total of 228 differentially expressed genes were obtained by screening of differential genes which 64 were up-regulated and 164 were down-regulated.Analysis of pathway expressions included mitosis,proliferation,migration and growth of high-density lipoprotein particle binding,ATP microtubule exercise,etc.,while signal function annotations mainly include metabolic pathway,cancer pathway,PI3K-Akt signaling pathway,cell cycle,p53 signaling pathway,PPAR signaling pathway,glycolysis/gluconeogenesis,tyrosine metabolism,etc;analysis of protein-protein interaction network diagrams,screening out the most representative seven genes(MAD2L1,ASPM,NUSAP1,BIRC5,DLGAP5,BUB1,ZWINT).GEPIA analysis results show that these seven genes are highly expressed in cancer tissues compared to normal tissues,and prognostic analysis also shows that all of them are related to recovery significant correlation.According to the CMAP analysis,10 small molecular compounds or drug candidates were selected.This study selected Resveratrol for subsequent experimental verification to explore its effect on A549 cells in vitro.Part 2: Effect of resveratrol on proliferation inhibition of lung adenocarcinoma A549 cells1.Resveratrol inhibits the proliferation of A549 cells: Verify the resveratrol inhibits the proliferation of A549 cells at different concentrations(12.5 μmol/L,25μmol/L,50 μmol/L,100 μmol/L,200 μmol/L),and set blank control and negative In the control group,the proliferation inhibition rate was detected by CCK-8 method after treatment with resveratrol for 24 h,48 h,and 72 h.The experimental results show that: at the same time and different concentrations,the resveratrol has a statistically significant inhibition effect on the growth of A549 cells(P<0.05),the greater the concentration,the higher the inhibition rate;the same concentration,there are statistics at different times difference(P<0.05),the longer the time,the higher the inhibition rate.2.The effect of resveratrol on glucose metabolism in A549 cells: Verified by glucose detection kit,resveratrol has an inhibitory effect,that the higher the concentration,the more significant,on glucose consumption in A549 cells.3.Effect of resveratrol on A549 cell migration: verify the effect of resveratrol at different concentrations(12.5 μmol/L,25 μmol/L,50 μmol/L,100 μmol/L,200μmol/L)on the migration of A549 cells Observe the healing of the scratches on the cells after 24 h and 48 h.The results showed that: compared with the control group,the higher the drug concentration,the lower the scratch healing rate,and compared with the control group,there were significant differences in the experimental group for 48 hours(P <0.05).Conclusion: In a certain concentration range,resveratrol can inhibit the proliferation of A549 cells,reduce the glucose consumption rate of A549 cells,also inhibit cell migration.