Expression pattern of Wif1 and β-catenin during development of anorectum in fetal rats with anorectal malformations

Abstract

PurposeThis study was performed to investigate the expression pattern of Wnt inhibitory factor 1 (Wif1) and β-catenin during anorectal development in normal and anorectal malformation (ARM) embryos and the possible role of Wif1 and β-catenin in the pathogenesis of ARM.MethodsARM was induced with ethylenethiourea on the 10th gestational day in rat embryos. Cesarean deliveries were performed to harvest the embryos. The expression pattern of Wif1 and β-catenin protein and mRNA was evaluated in normal rat embryos (n = 288) and ARM rat embryos (n = 306) from GD13 to GD16 using immunohistochemical staining, Western blot, and real time RT-PCR.ResultsImmunohistochemical staining revealed that in normal embryos Wif1 was constantly expressed in the cloaca from GD13 to GD16. On GD13 and GD14, Wif1-immunopositive cells were extensively expressed in the cloaca. On GD15, the expression of Wif1 were mainly detected on the very thin anal membrane. In ARM embryos, the epithelium of the hindgut and urorectal septum demonstrated faint immunostaining for Wif1 from GD14 to GD16. Western blot and real time RT-PCR revealed that Wif1 and β-catenin protein and mRNA expression level was significantly decreased in the ARM groups compared with the normal group on GD14 and GD15 (p < 0.05).ConclusionsThis study demonstrated that the expression pattern of Wif1 and β-catenin was disrupted in ARM embryos during anorectal morphogenesis, which demonstrated that downregulation of Wif1 and β-catenin at the time of cloacal separation into the primitive rectum and urogenital septum might related to the development of ARM.

Spatial transcriptomics reveals gene interactions and signaling pathway dynamics in rat embryos with anorectal malformation

“…According to previous theoretical foundations, this may be because of the continuous downward extension of the URS during GDs 14–15, causing the gradual separation of the hindgut and the urethra. This is a dynamic and continuous process, and regardless of whether the hindgut and urethra are successfully separated, this process ends before GD16 (Qi et al 2000 ; Tang et al 2018 ). Therefore, based on the characteristic module analysis of clusters 0–4, the gene modules that showed positive correlation trends at GD14 and GD15 may be the key effector molecules that promote normal or ARM development.…”

Identification of optimal endogenous reference RNAs for RT-qPCR normalization in hindgut of rat models with anorectal malformations

“…In previous ARM-related studies, Gapdh and Actb were most commonly used as reference genes in RT-qPCR (Geng et al, 2017; Tang et al, 2018; Xiao et al, 2018). However, this study revealed that these two genes could be replaced by better candidates.…”

Investigation of the Genome-Wide Genetic and Epigenetic Networks for Drug Discovery Based on Systems Biology Approaches in Colorectal Cancer

“…The next pathway is initiated with receptor WNT4. Several studies have shown that this pathway could inhibit the proliferation and migration of tumors (Seth and Ruiz I Altaba, 2016;Tang et al, 2018). In the WNT-signaling pathway, ligand WIF1 (signal transduction) binds with the receptor WNT4 to transmit the signal to TF ZEB1 through cascade proteins TUBA1B and TMEM39A.…”

“…In early-stage disease, PTK6 (hypoxic environment) binds with STAP2 to activate essential protein MAP4K1 and consequently the MAPK pathway (Fujita et al, 2014;Cho et al, 2017). WIF1 (Signal transduction) binds with WNT4 to activate the WNT signaling pathway (Seth and Ruiz I Altaba, 2016;Tang et al, 2018). MUC2 and HECTD3, modified by DNA methylation, could promote immune response, inflammation, cell cycle, and apoptosis.…”