Screening and expression analysis of the specific tissue and serum microRNA profile in Hirschsprung disease

Abstract

Objective To investigate the different expressions of pathological tissue and serum microRNAs(miRNAs) in Hirschsprung disease(HSCR). Methods Pathological colon tissues and serum samples were obtained from 52 confirmed HSCR cases respectively by surgery and pathology and from 52 matched controls, respectively.An initial screening of the tissues and serum microRNA expression were performed through TaqMan Low Density Array.The candidate tissue and serum miRNAs were validated by quantitative real-time-PCR in the 20 paired array samples and extra 32 paired samples after the integration of the screening result.The bioinformatical software online including miRbase, Target Scan, PicTar and MiRanda were used to predict the target mRNA of the consistent microRNAs in the tissues and the serum. Results Compared with the controls, 47 microRNAs were differently expressed in HSCR tissues, including 17 up-regulated miRNAs and 30 down-regulated miRNAs; 32 upregulated miRNAs were also detected to be differently expressed in the HSCR serum.Among these microRNAs, miR-218-1 and miR-885-5p were identified to have a consistent significant different expression in both tissues and the serum, which were validated as high-expressed in microarray samples and expanded 32 paired samples(miR-218-1: tissue array 0.017 58±0.002 29 vs 0.003 37±0.000 50, P<0.001; tissue expanded expression 0.013 53±0.001 74 vs 0.004 43±0.000 60, P<0.001.miR-885-5p: tissue array 0.000 30±0.000 11 vs 0.000 04±0.0000 08, P= 0.027 6; tissue expanded expression 0.004 59±0.000 16 vs 0.000 04±0.000 01, P=0.014 5.miR-218-1: serum array 0.769 60±0.285 50 vs 0.045 14±0.015 07, P=0.015 5; serum expanded expression 1.151 00±0.430 00 vs 0.023 07±0.003 81, P=0.008 7.miR-885-5p: serum array 1.595 00±0.441 70 vs 0.169 40±0.034 46, P=0.001 2; serum expanded expression 1.689 00±0.453 00 vs 0.146 10±0.031 24, P=0.001 2). Specifically, the target genes of these 2 microRNAs were RET, PLAG1 and NeuroD1, which had been reported to be directly related to HSCR. Conclusions Significantly differential expressed miRNAs exist in the pathological tissue and the serum of HSCR.MiR-218-1 and miR-885-5p, which showing consistent differential expression, may be involved in the pathogenesis of HSCR. Key words: microRNAs; Hirschsprung disease; Aganglionic colon; Serum