Zhang Bobin, Yang Jian, Song Aohua, Feng Wei, Guo Zhenhua
Ministry of Education Key Laboratory of Child Development, Chongqing Key Laboratory of Structural Birth Defect and Reconstruction, National Clinical Research Center for Child Health and Disorders, Department of Neonatal Surgery, Children's Hospital of Chongqing Medical University, 20, Jinyu Road, Yubei District, Chongqing, 400025, People's Republic of China.
Department of General Surgery, Yongchuan Maternal and Child Health Hospital, Chongqing, People's Republic of China.
Biochem Genet. 2024 Jun 7. doi: 10.1007/s10528-024-10852-z.
The objective of the study is to investigate how miR-146b-5p might contribute to the etiology of HSCR. The study investigated the expression levels of miRNA, mRNA, and proteins in colon tissues obtained from the HSCR and control groups. The role of miR-146b-5p in cell proliferation and migration was studied in vitro. The interaction between miR-146b-5p and RET was validated through a dual-luciferase reporter experiment. To assess the impact of miR-146b-5p on the development of the enteric nervous system, zebrafish embryos were micro-injected with either miR-146b-5p mimics or negative control, followed by subsequent evaluation. Compared to the control group, miR-146b-5p expression levels in the spastic region of HSCR were significantly increased. In vitro, miR-146b-5p prevented cell migration and proliferation by targeting RET pathway. In zebrafish, miR-146b-5p negatively regulates the migration of neural crest cells through a reduction in RET expression. Overexpression of miR-146b-5p hinders the development of mature neurons by decreasing RET expression. Additionally, the aberrant phenotypes induced by miR-146b-5p were partially ameliorated when RET mRNA was co-injected. By targeting RET in HSCR patients, aberrant expression of miR-146b-5p may play a unique role in the etiology of the disease and be involved in enteric nervous system development.
本研究的目的是探究miR-146b-5p如何对先天性巨结肠（HSCR）的病因产生影响。该研究调查了HSCR组和对照组结肠组织中miRNA、mRNA和蛋白质的表达水平。在体外研究了miR-146b-5p在细胞增殖和迁移中的作用。通过双荧光素酶报告基因实验验证了miR-146b-5p与RET之间的相互作用。为了评估miR-146b-5p对肠神经系统发育的影响，将miR-146b-5p模拟物或阴性对照显微注射到斑马鱼胚胎中，随后进行评估。与对照组相比，HSCR痉挛区域中miR-146b-5p的表达水平显著升高。在体外，miR-146b-5p通过靶向RET途径阻止细胞迁移和增殖。在斑马鱼中，miR-146b-5p通过降低RET表达负向调节神经嵴细胞的迁移。miR-146b-5p的过表达通过降低RET表达阻碍成熟神经元的发育。此外，当共注射RET mRNA时，miR-146b-5p诱导的异常表型得到部分改善。通过靶向HSCR患者中的RET，miR-146b-5p的异常表达可能在该疾病的病因中发挥独特作用，并参与肠神经系统的发育。
