Determination of miRNA profile alteration of host cells induced by enterovirus-A71 through high-throughput sequencing
10.3760/cma.j.cn112866-20211101-00188
- VernacularTitle:高通量测序确定肠道病毒A71型感染致宿主细胞miRNA表达谱的改变
- Author:
Pingping SUN
1
;
Xue LIU
;
Dan LI
;
Qing REN
;
Meng SU
;
Wenping GUO
;
Luanying DU
;
Jiangli WANG
;
Guangcheng XIE
Author Information
1. 承德医学院病原生物学教研室 病原防控研究室，承德 067000
- Keywords:
Enterovirus-A71;
Human tonsillar epithelial cells;
miRNA;
High-throughput sequencing
- From:
Chinese Journal of Experimental and Clinical Virology
2022;36(1):1-7
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To determine the alteration of miRNA profile of human tonsillar epithelial cells induced by enterovirus-A71 (EV-A71) infection.Methods:Human tonsillar epithelial cells UT-SCC-60B were infected with EV-A71 at multiplicities of infection (MOI) of 1 and total RNA was extracted using Trizol reagent. Small RNA library was constructed and high-throughput sequencing was performed using Illumina NextSeq 500. Differential significantly expressed known and novel miRNAs and putative targets were selected after the processing of raw data. Gene ontology (GO), kyoto encyclopedia of genes and genomes (KEGG) pathways were analyzed through online database. Kinds of miRNA could target EV-A71 genome was determined through psRNATarget. Validations of random selected miRNAs were done through real-time RT-PCR.Results:A total of 61 known significantly expressed miRNAs (21 miRNAs were down-regulated and 40 miRNAs were up-regulated) and 559 novel significantly expressed miRNAs were identified through high-throughput sequencing. Novel significantly expressed miRNA had typical "hairpin structure" of pre-miRNA. Fold changes of hsa-miR-517b-3p and hsa-miR-199a-5p which was determined by real-time RT-PCR had similar change trends with high-throughput sequencing. Putative targets of significantly expressed miRNA were referred to different biological processes and signaling pathways. A total of 24 significant miRNAs (5 known significantly expressed miRNAs and 19 novel significantly expressed miRNAs) had "seed sequence" in EV-A71 genome.Conclusions:Expression of miRNA profile in UT-SCC-60B was significantly changed by EV-A71 infection and the identified significantly expressed miRNAs potential target EV-A71 genome to regulate EV-A71 replication.
