Characteristics of the exosomal microRNA expression profile of HepG2 cells infected with Zika virus
10.3760/cma.j.cn112866-20220817-00180
- VernacularTitle:寨卡病毒感染人肝癌细胞HepG2后外泌体中差异miRNA分析
- Author:
Wankui ZHOU
1
;
Yue GONG
;
Jinbao GU
;
Yanhai WANG
Author Information
1. 广东省佛山市南海经济开发区人民医院检验科，佛山 528000
- Keywords:
Zika virus;
HepG2 cells;
Exosomes;
Exosomal miRNA;
Mosqutio brone disease
- From:
Chinese Journal of Experimental and Clinical Virology
2022;36(6):659-665
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To isolate and identify the exosomes from human hepatocellular carcinoma (HepG2) cells infected with Zika virus (ZKIV), and anaylize the profiles of exosomal miRNA in infectedand uninfected cells, predict and analyze their target genes.Methods:ZIKV(ZJ03 strain) was used to HepG2 cells at a multiplicity of infection (MOI) of 0.5. After 2 days, ultracentrifugation method were used to extract and purify the exosomes from ZIKV infected (Exo-ZIKV) and uninfected HepG2 cells (Exo-NC). The morphology and particle size of exosomes were observed by the transmission electron microscope; the size and concentration of exosomes were determined by Nanoparticle tracking analysis (NTA). Western blot was used to detect protein markers of exosomes. Small RNA was isolated from exosome and analyzed by small RNA deep sequencing. The differentially expressed miRNAs were further detected by bioinformatics analysis. KEGG and GO enriched analysis was used to analyze the target genes of differentially expressed miRNA.Results:Under the transmission electron microscopy, Exo-ZIKV and Exo-NC were spherical or cup-shaped bilayer membrane structures. Exo-ZIKV and Exo-NC shared the similar particle size distribution, wheara Exo-ZIKV displayed the increased concentration (24×10 8 paritcle/ml) than that of Exo-NC (3.06×10 8 paritcle/ml). Typical exosomal markers Hsp70, CD63 and CD9 were detected in both gourps. A total of 20 differentially expressed miRNAs (12 up-rugalated and 8 down-regulated miRNA) were detected. The target genes of differentially expressed miRNAs were suggested to be involved in the pyrimidine and purine metabolism pathways. Conclusions:ZIKV infection could promote the secretion of exosomes in HepG2cells. Furthermore, ZIKV infection cause the change of exosomal miRNA expression profile of HepG2 Cells. This study provides a basis for further elucidation of the role of exosomal miRNA in the mechanisms of ZIKV infection caused hepatocyte injury and provide new ideas for the treatment of ZIKV.
