RNA-Seq Data Analysis To The Mechanism Of IGFBP2 In Esophageal Adenocarcinoma Under Acid Bile Salt Environment

BackgroundEsophageal adenocarcinoma is a common malignant digestive tract tumor with poor prognosis,which seriously endangers human life and health.One of the most important risk factors for esophageal adenocarcinoma is gastroesophageal reflux disease,which runs through the whole process from Barrett’s esophagus to esophageal adenocarcinoma.The main components of reflux are gastric acid and acid bile salts(ABS).In recent years,it has been found that acid bile salts are more closely related to esophageal adenocarcinoma.It has been confirmed that acid bile salts can induce the expression of reactive oxygen species(ROS)in esophageal adenocarcinoma cells and cause DNA damage.IGFBP2 has been shown to be widely involved in multiple tumor-related regulatory pathways.Recent studies have found that IGFBP2 can attenuate ABS-induced DNA damage and apoptosis in esophageal adenocarcinoma,and is involved in regulating the chemotherapy resistance of esophageal adenocarcinoma.RNA sequencing technology(RNA-seq)can comprehensively obtain almost all transcript sequence information of a specific tissue in a specific state by measuring and comparing the sequence of transcripts,which is helpful to analyze the specific mechanism of IGFBP2 in esophageal adenocarcinoma.ObjectivesIn this study,esophageal adenocarcinoma cells were treated with ABS to simulate the physiological gastroesophageal reflux process.RNA-seq were used to analyze the transcriptome changes of esophageal adenocarcinoma after IGFBP2 silencing,aiming to find the potential targets and mechanisms of IGFBP2 in esophageal adenocarcinoma And a meta-analysis was performed to evaluate the effect of PPI use on BE,indirectly providing evidence that bile acids are a more important source of acid exposureMaterials and methods1.OE19 cell were treated with 100μM,200pM and 300μM ABS for 3h 6h and 12h,respectively.With the gene eypression level of IGFBP2 treated with OpM in Oh group as reference,the relative gene expression level of IGFBP2 in each group was determined by 2-AACT method,and the treatment groups with significant differences in expression were selected for subsequent experiments.2.The efficiency of three kinds of si-RNA silencing IGFBP2 was determined by 2-AACT method,and the most efficient si-RNA was selected for experiment.3.The OE19 cell line was treated with 200 μM ABS for 12h,and the OE19 gene expression profile after treatment was detected by RNA-seq.Bowtie2 software was used for gene comparison,RSEM software was used to calculate gene expression level,PossionDis method was used for differential expression analysis to screen differentially expressed genes,and GO and KEGG enrichment analysis were used to screen differentially expressed gene enrichment pathways.4.A systematic literature search was conducted in PubMed and other databases for studies on the relationship between PPI and BE progression.Literature was screened,and relevant data were extracted from the included studies for data summary and results analysis using Review Manage 5.2 software.Results1.Compared with Oh,the expression level of IGFBP2 in OE19 cells in most ABS treatment groups decreased significantly,and there was a very significant difference between 300μM and 6h groups.2.A total of 156 differentially expressed genes were screened out,including 62 up-regulated genes and 94 down-regulated genes.GO enrichment screened 12 significant differential pathways,and KEGG pathway enrichment screened 15 significant differential pathways.Conclusions1.The expression level of IGFBP2 in OE19 cells was correlated with the concentration and time of ABS stimulation.2.The top 10 up-regulated and down-regulated genes were not directly correlated with esophageal adenocarcinoma.Among them,ULK2,ACOT1 and FOXC2 are related to gastrointestinal tumors,which may become potential targets.3.The significantly differential enrichment pathways associated with esophageal adenocarcinoma mainly focused on response to hypoxia,Epithelial-mesenchymal transition,sex hormone metabolism and B vitamin metabolism.Response to hypoxia and epithelial-mesenchymal transition are closely related to IGFBP2,which may be the focus of future research on IGFBP2 and esophageal adenocarcinoma.4.PPI had no protective effect on the progression of BE patients.