The study of IncRNA LINC00265 in the Hirschsprung's disease at transcriptional level
Objective:To investigate the localization and expression of long non-coding RNA LINC00265 in the stenotic lesions and dilated normal intestine to provide new ideas for the pathogenesis of Hirschsprung's disease(HD).Methods:12 cases of HD from January 2017 to January 2019 were collected.The pathologically confirmed narrow full-thicknessintestine was used as the experimental group and the pathologically confirmed expended full-thickness intestine contatin-ing ganglion cells was used as the control group.The intestine was embedded in paraffin and sectioned routinely.Fluorescence in situ hybridiza-tion was used by adding hybridization solution containing probe CY3-labeled oligonucleotides to the sections,and the whole sections were scanned by scanner.The fields under the same multiple were selected for positive cell counting,and the final counting results were statistically analyzed.The target gene of LINC00265 were predicted by three prediction softwares of miranda,PITA and Targetscan.LINC00265 ceRNA network was constructed and screen out the genes associated with neu-rotrophin pathways and neuronal growth migration.Results:Fluorescence in situ hybridization results show that LINC002656 is mainly located in the cytoplasm of the mucosa and myometrial cells of the intestine,and the expression of LINC002656 in the stenotic lesions is significantly less than that of the normal intestine.The positive cell counts at same multiple of the stenotic lesions and normal intestinal were 3.67±1.30 & 10.33±2.46 respectively with different statistically significant(P＜0.05),and the related down-regulated target genes were 97,of which angiotensin-converting enzyme,guanine nucleotide exchange factor protein,and microtubule movement protein were associated with brain-derived neurotrophic factor(BDNF).Conclusions:This experiment shows that LINC00265 may regulate the microenviroment of the intestinal wall by regulating the expression of brain-derived neurotrophic factor and interfere the migration and proliferation of intestinal neural crest stem cells,which may berelatedto the occurrence of HD.
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