环状RNA及Wnt3a在肛门直肠畸形发生发展中的作用及机制的研究

先天性肛门直肠畸形(anorectal malformation ARMs)的基因表达调控机制复杂，涉及多种信号通路。课题组前期工作已证实Wnt通路为调控ARMs发生的主要途径之一 ，同时证实Wnt3a与ARMs的发生及进一步的病理改变密切相关，经生物学预测分析在其表达过程中涉及到mir-128-1/mir-216a及多种环状RNA（circRNAs），但其具体的调控机制仍然未知。本课题拟利用乙烯硫脲致畸的ARMs大鼠模型、肠神经干细胞系，Wnt3a的3’UTR荧光素酶表达载体，利用RT-PCR、Western-blot、激光捕获以及跨胎盘siRNAs干扰等技术，验证mir-128-1/mir-216a及多种circRNAs与Wnt3a之间的相互调控作用及对ARMs发生的调控机制，为ARMs的发病调控机制绘制出更全面、详细的网络框架及开拓疾病新的治疗思路。

Anorectal malformations(ARMs) are one of the most common congenital anomalies in the newborn. The gene regulation mechanism associated with ARMs is complicated. On the basis of our previous research, the Wnt is one of the main pathways in ARMs .Wnt3a is an important signal gene in classical pathway and the more detailed regulatory mechanisms of Wnt3a in ARMs are still not clear. We have confirmed much circRNAs and mir-128-1/mir-216a are closely related with Wnt3a by forecast using Targetscan and Starbasev2.0 software.. The design of our study is to use ethylenethiourea (ETU)-exposed rat embryos with ARMs，enteric neural stem cell and the virus vectors of Wnt3a’s 3’UTR with immune luciferase. The effect and regulation mechanism of circRNAs will be confirmed by using real-time PCR (RT-PCR), western blot and trans- planceta interference of siRNAs to test the relationship between mir-128-1/mir-216a、circRNAs and ARMs and their regulation mechanism. . This study will give a new point of view to learn the mechanism of circRNAs and Wnt3a in ARMs and more comprehensive understanding the pathogenesis of circRNAs and Wnt3a in the ARMs.This will concentrate to guide the ARMs’ clinical treatments.