Wang Juping,Yan Yingnan,Zhao Fangzheng,et al.Molecular mechanism of hsacirc_049751 influencing the pathogenesis of Hirschsprung’s disease via neural cell migration[J].Journal of Clinical Pediatric Surgery,2026,(06):552-557.[doi:10.3760/cma.j.cn101785-20251112-00061]
hsacirc_049751通过调控神经细胞迁移影响先天性巨结肠发病进程的分子机制研究
- Title:
- Molecular mechanism of hsacirc_049751 influencing the pathogenesis of Hirschsprung’s disease via neural cell migration
- Keywords:
- Hirschsprung Disease; Sequence Analysis; RNA; Circular RNA; Glial Cell Line-derived Neurotrophic Factor; Enteric Nervous System
- 摘要:
- 目的 探讨环状 RNA hsacirc_049751 在先天性巨结肠(hirschsprung disease,HSCR)中的表达特征并初步研究其潜在分子调控机制。方法 采用Illumina HiSeq平台对HSCR与正常肠组织进行RNA测序(RNA-seq),筛选差异表达circRNAs。通过qRT-PCR在扩大样本量中验证候选circRNAs的表达水平。利用生物信息学方法预测hsacirc_049751的潜在结合miRNA及其下游靶基因。同时,在SH-SY5Y细胞中过表达或敲低hsacirc_049751,通过细胞愈合实验(wound healing assay,WHA)检测其对神经细胞迁移过程的影响。结果 HSCR组织中存在显著的circRNAs表达异常。qRT-PCR验证结果显示,hsacirc_049751在HSCR组织中的表达水平显著低于正常对照组(P<0.01)。功能实验表明,过表达hsacirc_049751可显著促进SH-SY5Y细胞迁移,而抑制hsacirc_049751则显著阻碍细胞迁移。生物信息学分析预测,hsacirc_049751可能作为竞争性内源RNA吸附miR-516b,进而影响其对胶质细胞系来源的神经营养因子(glial cell line-derived neurotrophic factor,GDNF)及其受体家族成员α3(glial cell line-derived neurotrophic factor family receptor alpha-3,GFRA3)的调控,从而影响肠神经系统发育相关通路。结论 hsacirc_049751在HSCR中显著低表达,其表达水平与miR-516b及GDNF/GFRA3信号通路相关,提示其可能参与肠神经系统发育过程,为进一步阐明HSCR的分子调控机制及寻找潜在诊疗靶点提供了新的理论依据。
- Abstract:
- Objective To explore the expression profile of circular RNA hsacirc_049751 in Hirschsprung’s disease (HSCR) tissues and preliminarily examine its potential molecular regulatory mechanisms. Methods High-throughput RNA sequencing (Illumina HiSeq) was performed on intestinal tissues from HSCR children and normal controls to identify differentially expressed circRNAs.The expression of hsacirc_049751 was further validated in an expanded cohort by quantitative real-time polymerase chain reaction (qRT-PCR).Bioinformatic analyses were conducted for predicting potential miRNAs interacting with hsacirc_049751 and their downstream target genes.Functional assays were performed in SH-SY5Y cells through an over-expression or a knockdown of hsacirc_049751.And its effects on neural cell migration were evaluated by wound healing assay (WHA). Results Multiple circRNAs became significantly dysregulated in HSCR tissues.qRT-PCR validation demonstrated that hsacirc_049751 was markedly down-regulated in HSCR as compared with normal controls (P<0.01).Functional assays indicated that an over-expression of hsacirc_049751 greatly enhanced SH-SY5Y cell migration,whereas an inhibition of hsacirc_049751 significantly suppressed the migratory capacity of these cells.Bioinformatic predictions revealed that hsacirc_049751 might function as a competing endogenous RNA by sponging miR-516b,thereby attenuating its inhibitory effects on glial cell line-derived neurotrophic factor (GDNF) and glial cell line-derived neurotrophic factor family receptor alpha 3 (GFRA3).Both are key components of enteric nervous system development-related signaling pathways. Conclusions Hsacirc_049751 becomes significantly down-regulated in HSCR and may influence enteric nervous system development through regulating the axis of miR-516b-GDNF/GFRA3 signaling.These findings provide novel mechanistic insights into the molecular pathogenesis of HSCR and hint at potential diagnostic and therapeutic targets.
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备注/Memo
收稿日期:2025-11-12。
基金项目:上海市自然科学基金(22ZR1451500)
通讯作者:王阳,Email:wangyangwy@shsmu.edu.cn