Differential expression of microRNA in neonatal necrotizing enterocolitis based on high-throughput sequencing

Abstract

Objective To analyze the differential expression of microRNA (miRNA) and its significance in patients with neonatal necrotizing enterocolitis (NEC). Methods Twenty-five patients diagnosed with NEC with Bell stage ≥Ⅱ, and 25 non-NEC patients as control group admitted to Guangzhou Women and Children's Medical Center between October 2014 and November 2015 were collected. White blood cells were extracted from the peripheral blood. Five samples were selected randomly each from NEC group and control group, and sequenced by second-generation Illumina high-throughput sequencing, screened for differentially expressed miRNA and analyzed for target genes prediction and biological function. The rest samples of the two groups were detected by real-time fluorescent quantitative polymerase chain reaction technology (RT-qPCR), the results were used to validate the results of high-throughput sequencing. Differentially expressed miRNA in the two groups of data was analyzed using DEGseq software. P<0.05 was considered statistically significant. P<0.01, q<0.001 and丨Log2 Ratio丨≥1 were taken as criteria for screening the differential expression. The differential expressions of miRNA in NEC group and control group were analyzed by cluster analysis using MeV4.6 software. Results A total of 482 miRNAs were differentially expressed in the two groups, with significant difference (P<0.05). Among them, 126 were known miRNAs with significantly differential expression in the two groups, with 58 being up-regulated and 68 being down-regulated. The results of up-regulated miRNAs (hsa-miR-223-5p, -183-3p, -222-5p) and down-regulated miRNAs (hsa-miR-23b-5p, -150-5p, -146a-3p, -1298-5p) were confirmed to be consistent with the results of sequencing. Bioinformatics analysis showed that the target genes with differential miRNA expression mainly involved Toll-like receptor signal transduction pathway, mitogen-activated protein kinase pathway, JAK-STAT and other signal transduction pathways. Conclusions There are significantly differential expressions of miRNAs in peripheral white blood cells of NEC neonates. These miRNs may be involved in the occurrence and development of NEC via adjusting different target genes to regulate the signal pathway. Key words: Enterocolitis, necrotizing; MicroRNAs; High-throughput nucleotide sequencing; Infant, newborn