miR-141-3p affects β-catenin signaling and apoptosis by targeting Ubtd2 in rats with anorectal malformations

Area of Science:

• Developmental Biology
• Molecular Biology
• Genetics

Background:

• Anorectal malformations (ARMs) represent the most frequent congenital gastrointestinal anomalies.
• Understanding the molecular mechanisms underlying ARM pathogenesis is crucial for improving diagnostic and therapeutic strategies.

Purpose of the Study:

• To investigate the role of miR-141-3p and its target gene, Ubtd2, in the development of anorectal malformations.
• To elucidate the molecular pathways involving miR-141-3p, Ubtd2, USP5, and beta-catenin in intestinal epithelial cells.

Main Methods:

• Induction of an ARM rat model using ethylenethiourea.
• Detection of miR-141-3p and Ubtd2 expression using fluorescence in situ hybridization and immunofluorescence.
• Dual-luciferase reporter assay to confirm the targeting relationship between miR-141-3p and Ubtd2.
• Western blotting, co-immunoprecipitation, flow cytometry, and caspase 3/7 staining to analyze protein interactions and cell apoptosis.

Main Results:

• miR-141-3p was significantly downregulated in the ARM rat model, while Ubtd2 expression was increased.
• Downregulation of miR-141-3p promoted apoptosis in intestinal epithelial cells (IEC-6) via Ubtd2.
• Ubtd2 affected the protein expression of USP5 and beta-catenin, with USP5 binding to both Ubtd2 and beta-catenin.
• Aberrant apoptosis and dysregulated beta-catenin signaling were observed in the context of hindgut development.

Conclusions:

• Targeting of Ubtd2 by miR-141-3p plays a critical role in regulating intestinal epithelial cell apoptosis.
• The miR-141-3p/Ubtd2 axis influences beta-catenin signaling, potentially contributing to the pathogenesis of anorectal malformations.
• These findings provide novel insights into the molecular basis of ARMs and suggest potential therapeutic targets.