Tang Weibing, Tang Junwei, He Jun, Zhou Zhigang, Qin Yufeng, Qin Jingjing, Li Bo, Xu Xiaoqun, Geng Qiming, Jiang Weiwei, Wu Wei, Wang Xinru, Xia Yankai
State Key Laboratory of Reproductive Medicine, Institute of Toxicology, School of Public Health, Nanjing Medical University, Nanjing, China.
Laboratory of Modern Toxicology (Nanjing Medical University), Ministry of Education, China.
J Cell Mol Med. 2015 Jun;19(6):1197-207. doi: 10.1111/jcmm.12454. Epub 2015 Mar 19.
Hirschsprung's disease (HSCR) is a rare congenital disease caused by impaired proliferation and migration of neural crest cells. We investigated changes in expression of microRNAs (miRNAs) and the genes they regulate in tissues of patients with HSCR. Quantitative real-time PCR and immunoblot analyses were used to measure levels of miRNA, mRNAs, and proteins in colon tissues from 69 patients with HSCR and 49 individuals without HSCR (controls). Direct interactions between miRNAs and specific mRNAs were indentified in vitro, while the function role of miR-218-1 was investigated by using miR-218 transgenic mice. An increased level of miR-218-1 correlated with increased levels of SLIT2 and decreased levels of RET and PLAG1 mRNA and protein. The reductions in RET and PLAG1 by miR-218-1 reduced proliferation and migration of SH-SY5Y cells. Overexpression of the secreted form of SLIT2 inhibited cell migration via binding to its receptor ROBO1. Bowel tissues from miR-218-1 transgenic mice had nerve fibre hyperplasia and reduced numbers of gangliocytes, compared with wild-type mice. Altered miR-218-1 regulation of SLIT2, RET and PLAG1 might be involved in the pathogenesis of HSCR.
先天性巨结肠症（HSCR）是一种由神经嵴细胞增殖和迁移受损引起的罕见先天性疾病。我们研究了HSCR患者组织中微小RNA（miRNA）及其调控基因的表达变化。采用定量实时聚合酶链反应和免疫印迹分析来检测69例HSCR患者和49例非HSCR个体（对照）结肠组织中miRNA、信使核糖核酸（mRNA）和蛋白质的水平。在体外确定了miRNA与特定mRNA之间的直接相互作用，同时通过使用miR-218转基因小鼠研究了miR-218-1的功能作用。miR-218-1水平升高与SLIT2水平升高以及RET和PLAG1 mRNA及蛋白质水平降低相关。miR-218-1导致的RET和PLAG1水平降低减少了SH-SY5Y细胞的增殖和迁移。分泌形式的SLIT2过表达通过与其受体ROBO1结合抑制细胞迁移。与野生型小鼠相比，miR-218-1转基因小鼠的肠组织出现神经纤维增生且神经节细胞数量减少。miR-218-1对SLIT2、RET和PLAG1的调控改变可能参与了HSCR的发病机制。
