Expression Levels And Significance Of The Related MiRNAs In Colonic Tissues From Children With Hirschsprung's Disease

Background:Hirschsprung's disease(HSCR),is a common digestive system malformation in pediatric general surgery.The etiology of Hirschsprung's disease is the absence of ganglion cells in the submucosal and myenteric plexi of the gut.It results in the intestinal spasms and functional obstruction.The high-throughput screening was used to select a subset of differentially expressed miRNAs between HSCR stenotic colon segments and normal colon segments including miRNA-147?miRNA-1303?miRNA-320 and miRNA-133b.In this study,TapMan probe qRT-PCR verifiaction was used to detect the expression levels of relative miRNAs in colon tissues.Aims:To investigate the differentially expressed miRNAs between Hirschsprung's disease stenotic colon segments and normal colon segments.Methods:A total of 65 human colon tissues including 33 HSCR stenotic colon segments and 32 matched normal colon segments who undertook operative treatment(such as enterostomy and colectomy)without congenital enteric nerve disease were collected from Children's Hospital of Soochow University.The total RNA of colon tissues was extracted and purified by TRIzol.cDNA was formed by reverse transcriptase.TaqMan probe qRT-PCR was used to verify the expression levels of relative miRNAs in colon tissues.microT-CDS database and TargetScan database were used to screen out the target of relative miRNAs.Gene ontology and KEGG were used to analyze these target genes.Results:Decreased expression of miRNA-147 was verified by qRT-PCR and statistical analysis(the Student's t-test)in HSCR colon tissues compared to that in normal colon tissues(p<0.0001),the same as miRNA-1303(p=0.0052).The area under the ROC curve(AUC)of miRNA-1303 was 0.6998(95%CI:0.5693,0.8304;p=0.0056),and that of miRNA-147 was 0.8258(95%CI:0.7236,0.9280;p<0.0001).MiRNA-320(p=0.0894)was proved to have tiny differences without statistical significance,whose AUC was 0.6231(95%CI:0.4848,0.7614;p=0.0880).miRNA-133b(p=0.1229)was found no significant difference between Hirschsprung's disease stenotic colon segments and normal colon segments.The AUC of miRNA-133b was 0.6117(95%CI:0.4701,0.7534;P=0.1215).GO enrichment analysis and KEGG indicated that these intersectional target genes of miRNA-1303 and miRNA-147 were obviously enriched in biological regulation,protein binding and Hippo signaling pathway.Conclusions:The over-expressed miRNA-147 and miRNA-1303 may take part in the pathogenesis of HSCR.The role of miRNA-320 in the pathogenesis of HSCR has not been determined.MiRNA-133b does not participate in the pathogenesis of HSCR.