Expression analysis of miRNA profiles in apheresis platelets at the end of storage
10.13303/j.cjbt.issn.1004-549x.2021.08.005
- VernacularTitle:贮存末期血小板miRNA的表达谱分析
- Author:
Mingming QIAO
1
;
Wei LI
2
;
Yunlong ZHUANG
1
;
Hui YE
1
;
Qun LIU
1
;
Xia GAI
1
;
Yuanfeng CHEN
1
;
Hua SHEN
1
;
Baoyun JIANG
1
;
Yuxia WANG
1
Author Information
1. Blood Center of Shandong Province, Jinan 250014, China
2. Heze Central Blood Station
- Publication Type:Journal Article
- Keywords:
microRNA, platelet;
miRNA expression profile;
DNA Nano Ball (DNB) sequencing;
functional analysis;
fluorescence quantitative PCR;
small RNA
- From:
Chinese Journal of Blood Transfusion
2021;34(8):821-827
- CountryChina
- Language:Chinese
-
Abstract:
【Objective】 To investigate the changes of platelet microRNA (miRNA) expression profiles of storage in vitro, and to explore the molecular mechanism of miRNAs involved in the regulation of platelet storage lesion (PSL). 【Methods】 20 platelet samples (5 mL / sample) were collected from apheresis platelet donors, fully mixed and stored in a shaker with (22±2) ℃ horizontal agitation, sampled on day 1 and day 5, and sequenced by DNA nanoball (DNB) sequencing technology. The miRNAs with more than 2 times expressions (P<0.01) were considered as significantly differences between d5 and d1 groups. The miRanda and TargetScan softwares were used to predict the target genes. Gene Ontology (GO) function enrichment analysis and Kyoto Encyclopedia of genes and genomes (KEGG) pathway enrichment analysis were performed on the target genes of significant differentially expressed miRNAs. The expression of miRNAs was verified by real-time fluorescence quantitative PCR (qRT-PCR). 【Results】 Compared with d1 group, 315 miRNAs with significantly different expression (P<0.01) were screened in d5 group, including 146 up-regulated miRNAs (such as miR-146a, let-7b), and 169 down-regulated miRNAs (such as mir-30d, mir-142). Among 126 known miRNAs, 43 were up-regulated and 83 were down-regulated. There are 189 new miRNA sequences. The enriched GO terms of target genes of differentially expressed miRNAs in d5 and d1 groups included cell components, organelles, cell membrane and other cell structures, molecular functions such as adhesion, catalysis and activity, and biological processes such as cell processing, metabolism, biological regulation and stress. The corresponding pathways in the top 10 of KEGG enrichment were mainly signal transduction, secretion, membrane transport, amino acid metabolism, polysaccharide metabolism, protein synthesis and environmental adaptation. The 6 randomly selected differentially expressed miRNAs verified by qRT-PCR were consistent with those of DNB sequencing. 【Conclusion】 The expression profiles of platelets miRNAs have changed significantly between the d1 and d5 of storage in vitro. Functional prediction suggested that these miRNAs might be involved in the regulation of platelet PSL.
