陆军军医大学学报 (Jul 2023)
Role of ARRDC3 in enterovirus D68-infected A549 cells
Abstract
Objective To explore the expression of a-arrestin domain containing protein 3 (ARRDC3) in EV-D68 infected A549 cells and its effect on the biological behaviors of the cells. Methods The samples before and after EV-D68 infecting host cells (A549 cells) were analyzed by high-throughput whole transcriptome analysis with sequencing. After bioinformatics analysis was used to screen differentially expressed genes (DEGs), RT-qPCR and Western blotting were used to verify the expression of ARRDC3 at mRNA and protein levels. Then the A549 cells were designed and divided into 4 groups: control group (A549 cells not infected with EV-D68), infection group (A549 cells infected), knockdown ARRDC3 infection group (ARRDC3 siRNA interference-mediated knockdown in A549 cells after infection), knockdown control infection group (A549 cells infected with EV-D68 transfected with NC siRNA interference). Flow cytometry, CCK-8 assay, and Transwell assay were used to determine the effects of ARRDC3 on cell cycle, proliferation, and migration in above groups of cells. Results Gene sequencing showed there were 239 DEGs with more than 2-fold differential expression, with 135 up-regulated and 104 down-regulated genes, and ARRDC3 is one of DEGs. After A549 cells were infected with EV-D68, the expression level of ARRDC3 was significantly increased through RT-qPCR and Western blotting (P < 0.05), cell cycle was arrested and cell migration was inhibited. Silencing of ARRDC3 through siRNA transfection improved EV-D68 infection to A549 cells and prolonged the cell cycle arrested at DNA synthesis phase (P < 0.05). Although the silencing showed no significant effect on the proliferation of host cells, cell migration ability was obviously enhanced (P < 0.05). Conclusion ARRDC3 is significantly up-regulated after EV-D68 infects A549 cells, and exerts its influence on host cells by blocking the DNA synthesis phase and inhibiting migration of host cells.
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