High-throughput sequencing reveals the dynamic changes in the differential expression of cellular miRNAs during EBV reactivation
10.3760/cma.j.cn112866-20240930-00147
- VernacularTitle:高通量测序揭示EBV再激活过程中宿主细胞miRNA差异表达的动态变化
- Author:
Haotian LI
1
;
Hui WANG
1
;
Jiao WANG
1
;
Xuexin LU
1
;
Jieqiong ZHANG
1
;
Mingming WANG
1
;
Dongbo YU
1
;
Ying LI
1
;
Shiwen WANG
1
Author Information
1. 中国疾病预防控制中心病毒病预防控制所，北京 102206
- Publication Type:Journal Article
- Keywords:
Epstein-Barr virus;
Raji cell line;
MiRNA;
High-throughput sequencing
- From:
Chinese Journal of Experimental and Clinical Virology
2025;39(1):1-8
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the dynamic changes of cellular miRNA expression profiles in EBV latently infected Raji cells upon reactivation with Phorbol ester (TPA).Methods:Total RNA was extracted using TRIzol reagent from Raji cells treated with TPA at different time points (0 h, 24 h, 48 h). Small RNA libraries were constructed and sequenced on an Illumina SE50 platform. Differentially expressed miRNAs were identified, and their target genes were predicted and functionally annotated. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were carried out through online tools. Additionally, miRanda and RNAhybrid software were used to predict cellular miRNAs targeting the EBV genome. Real-time RT-qPCR was employed to validate the expression levels of differentially expressed novel miRNAs.Results:High-throughput sequencing identified 1 301 celluar miRNAs, comprising 1 189 known and 112 novel miRNAs. A total of 264 known differentially expressed cellular miRNAs and 13 novel miRNAs were identified through high-throughput miRNA sequencing. Secondary structure prediction revealed that the novel miRNAs exhibited typical pre-miRNA hairpin structures. Stem-loop quantitative real-time PCR (RT-qPCR) validation of Novel_miR_183 and Novel_miR_242 did not exhibit a statistically significant difference ( F=1.407, P=0.370 7 for Novel_miR_183; F=1.277, P=0.397 0 for Novel_miR_242) between the TPA-stimulated and untreated groups. Target gene prediction analysis revealed that the differentially expressed cellular miRNAs were involved in various important biological processes and signaling pathways. Furthermore, 1 189 known cellular miRNAs and 108 novel miRNAs were predicted to target the EBV genome. Conclusions:Treatment of Raji cells with TPA stimulation successfully reactivated Raji cells and significantly altered their miRNA expression patterns. Differentially expressed miRNAs were identified, suggesting that these miRNAs probably play crucial roles in regulating EBV infection and replication by directly targeting the EBV genome.
