Analysis of the Promotion with Reactive Oxygen Species on Migration of Imma-ture Dendritic Cells Based on RNA-seq Technique
Based on the detection of the cell cycle and migration ability of immature dendritic cells(imDCs)in response to reactive oxygen species(ROS),this study aims to analyze differentially expressed genes(DEGs)through RNA sequencing(RNA-seq)techno-logy and explore the potential mechanism of ROS promoting the migration of imDCs.The oxidative stress model of imDCs was construc-ted by simulating ROS with H2O2.The CCK8 method was used to detect cell viability.The flow cytometry was deployed to analyze cell apoptosis and the changes of cell cycle.The free migration of imDCs was analyzed by cell real-time imaging.Total RNA was extracted and RNA-seq was performed.GO and KEGG enrichment analysis of DEGs was performed.Transcripts related to metabolism and skele-ton regulation were screened according to the changes of DEGs expression|log2FC|≥1 and FDR≤～0.01.The result showed that there was no significant changes in cell viability and apoptosis of imDCs when H2O2 concentration was less than 100 μmol/L.The cell cycle of imDCs was arrested at G0/G1 phase and the free migration ability of imDCs was enhanced when in the ROS environment.RNA-seq data showed there were 282 DEGs between the control group and the H2O2 treatment group.Among them,225 differentially expressed genes were down-regulated,and 57 were up-regulated.GO and KEGG enrichment analyses found that these genes were involved in me-tabolism,cell cycle and cytoskeleton-related signalling pathways.Gsta1,Gsta3,Cstdc5 and Prdx1 genes involved in response to xeno-biotic metabolism were up-regulated but Shmt2 was down-regulated.S100a8,Ppbp and Tm4sf19 genes involved in skeleton regulation were also up-regulated.Simultaneously,Nusap1,Stmn1,Kif20 and Prc1 were down-regulated.Therefore,the activation of the antioxi-dant system,cytoskeletal remodeling and cell cycle arrest may be involved in regulating the migration ability of imDCs in response to extracellular ROS.
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