LncRNA-RMST/SOX2/miR-1251影响肠神经嵴细胞迁移和增殖在先天性巨结肠发生中的机制研究

Hirschsprung’s disease (HSCR)is highly associated with the failure of migration of enteric neural crest-derived cells (ENCCs). Using lncRNA microarray analysis of the established HSCR samples, together with cell experiment results, we found the expressions of lncRNA-RMST, miR-1251 and SOX2 decreased significantly. In vitro, results showed that downregulations of lncRNA-RMST and miR-1251 inhibited migration and proliferation of neuron. Besides, bioinformatics prediction showed that lncRNA-RMST interacting with SOX2 which combined with the promoter region of miR-1251, participated in influence of the ENCCs migration. We hypothesized that lncRNA-RMST is required for the binding of SOX2 to promoter regions of miR-1251 and coregulates the target gene AHNAK, causing the failure of ENCCs’ migration to intestine and finally leading to HSCR..Our research aims to investigate the role of LncRNA-RMST/SOX2/miR-1251 in the failure of ENCCs migration and proliferation. We will identify its role in the occurrence of HSCR in vitro experiments. This research will bring new viewpoints to the HSCR pathogenesis, providing potential targets in the prevention and treatment of HSCR during the embryonic period.

先天性巨结肠（HSCR）发生的主要原因是胚胎期肠神经嵴细胞（ENCCs）迁移增殖障碍。我们采用lncRNA芯片分析，整合人群样本及细胞检测发现：HSCR病变肠管中lncRNA-RMST及miR-1251及转录因子SOX2均为低表达状态，体外实验低表达lncRNA-RMST及miR-1251均抑制神经细胞迁移及增殖，结合生物信息学预测SOX2可以与lncRNA-RMST相结合，同时与miR-1251启动子区域结合，共同参与ENCCs迁移增殖障碍，由此假设：lncRNA-RMST作为SOX2转录共调控因子通过miR-1251调控靶基因AHNAK表达，影响ENCCs的迁移和增殖，参与先天性巨结肠发生。.本研究将阐述LncRNA-RMST/SOX2/miR-1251通路影响ENCCs迁移和增殖，探讨该通路在HSCR发生中的作用，本研究可为HSCR发生机制提供新视点和胚胎期防治提供潜在靶标。

在胚胎期肠神经嵴细胞（Embryonic intestinal neural crest cells, ENCCs）向消化道远端的迁移和增殖出现障碍是导致先天性巨结肠(Hirschsprung’s disease，HSCR)发生的主要原因，但其具体的发生机制仍不清楚。.在本项研究中，我们利用长链非编码RNA（long none-coding RNA, lncRNA）芯片筛选到与神经发育密切相关的lncRNA -RMST在HSCR患儿病变肠管中的表达明显低于对照组织，生物信息学分析提示miR-1251是存在于RMST内含子区域的miRNA。进一步扩大组织样本进行验证发现RMST、miR-1251在HSCR病变段表达显著降低，体外实验提示下调RMST或miR-1251的表达明显抑制神经细胞的迁移增殖功能。.然而，RMST并不直接通过影响miR-1251的表达行使功能。通过生物信息学分析，我们推测miR-1251受到转录因子SOX2调控，而RMST可结合SOX2进而加强SOX2对miR-1251的调控作用，体外实验则证实在同时下调RMST与SOX2后，miR-1251的表达较单独下调SOX2减少更多。最后，我们证实AHNAK是miR-1251的靶基因，而细胞实验结果表明miR-1251是通过下调AHNAK影响神经细胞的增殖及迁移。.本研究率先揭示了RMST作为SOX2转录共调控因子，协同调控miR-1251，使得靶基因AHNAK的表达上调，抑制神经嵴细胞的增殖与迁移的机制。RMST/SOX2/miR-1251/AHNAK通路的提出也为HSCR发生机制的探寻提供了新的线索，这对HSCR的防治具有一定的理论指导意义。