MiR‐141‐3p affects β‐catenin signaling and apoptosis by targeting Ubtd2 in rats with anorectal malformations. Issue 1 (3rd November 2022)

• Record Type: Journal Article
• Authors: Wang, Chen Yi; Li, Si Ying; Xiao, Yun Xia; Zhen, Lin; Wei, Xiao Gao; Tang, Xiao Bing; Yuan, Zheng Wei; Bai, Yu Zuo
• Is Part Of: Annals of the New York Academy of Sciences. Volume 1518:Issue 1(2022)
• Page Start: 315
• Page End: 327
• Publication Date: 2022-11-03
• Subjects: anorectal malformations -- apoptosis -- embryonic development -- hindgut -- miR‐141‐3p; Medical sciences -- Periodicals; Medicine -- Periodicals; Science -- Periodicals; 610
• DOI: 10.1111/nyas.14924
• Languages: English
• ISSNs: 0077-8923

Abstract

Abstract: Anorectal malformations (ARMs) are the most common gastrointestinal malformations. miR‐141‐3p was obtained from whole‐transcriptome sequencing, and Ub domain‐containing protein 2 ( Ubtd2 ) was predicted as the target gene. An ARM rat model was induced using ethylenethiourea. Fluorescence in situ hybridization and immunofluorescence were used to detect the spatiotemporal expression of miR‐141‐3p and Ubtd2, respectively. A dual‐luciferase reporter assay confirmed their targeting relationship, and cell proliferation and apoptosis were investigated after transfection in the intestinal epithelium (IEC‐6). Additionally, western blotting and co‐immunoprecipitation were used to examine the protein levels and the endogenous binding relationship. miR‐141‐3p was downregulated in the ARM group, whereas Ubtd2 increased and colocalized with TUNEL‐positive cells. After miR‐141‐3p inhibition, protein expression of USP5 and β‐catenin was affected via Ubtd2, and USP5 could bind to both Ubtd2 and β‐catenin. Flow cytometry analysis and caspase 3/7 staining demonstrated that downregulated miR‐141‐3p promoted cell apoptosis through Ubtd2. In summary, targeting Ubtd2 decreased in miR‐141‐3p and promoted apoptosis of intestinal epithelium and regulated β‐catenin expression. This may cause aberrant apoptosis during hindgut development and mediate the imbalance of β‐catenin signaling in the cloaca, further affecting the occurrence of ARMs. Abstract : Anorectal malformations (ARMs) are the most common gastrointestinal malformations. This study aimed to investigate the target relationship between Ub domain‐containing protein 2 ( Ubtd2 ), which we show by whole‐transcriptome sequencing is upregulated in the hindgut of ARM rat fetuses, and miR‐141‐3p, which is predicted to bind the Ubtd2 mRNA. Furthermore, we evaluated the effects of miR‐141‐3p and Ubtd2 on rat intestinal epithelial cell proliferation, apoptosis, and β‐catenin signaling.