关键词: 肠道病毒A型, 人, 转录组, 基因表达, 自噬, 横纹肌肉瘤细胞, 免疫应答

Abstract: Objective　To investigate the effect of EV71 infection on autophagy and immune response of human rhabdomyosarcoma (RD) cells and its mechanism based on transcriptome sequencing. Methods　Multiplicity of infection (MOI) was determined with different doses of EV71 infected RD cells. When MOI=1, the different time points (0, 0.5, 1, 1.5, 3, 6, 9 and 12 h) were selected for follow-up experiments. The changes in cell morphology and cell number were observed by inverted microscope. Western blot assay was used for observing protein expression. Changes in cell function and gene expression were analyzed by transcriptome sequencing. The intracellular protein expression was observed by high-conglutinative cell imaging. Results　The inverted microscope showed that cells gradually changed from fusiform adherence to circular suspension at 3 h after virus infection, and the number of cells decreased significantly compared with the 0 h group (P＜0.05). Western blot results showed that the cells had synthesised viral capsid protein1 (VP-1) at 6 h after infection, and values of LC-3Ⅱ/LC-3Ⅰwere significantly increased at 9 h after infection compared with those in the 0 h group (P＜0.01). Transcriptome sequencing results showed that gene expression changes in cells reached the peak at 3 h after infected, with      1 199 up-regulated genes and 126 down-regulated genes. KEGG analysis suggested that most of the altered signaling pathway molecules were related to autophagy and immunity. High-conglutinative cell imaging showed that the number of cells decreased and the average fluorescence intensity of intracellular autophagy protein increased after infection (P＜0.05). Conclusion　EV71 infects RD cells to promote the replication and assembly of the virus in cells through the autophagy related signaling pathway.

Key words: enterovirus A, human, transcriptome, gene expression, autophagy, rhabdomyosarcoma cell, immune response