Downregulation of microRNA-483-5p Promotes Cell Proliferation and Invasion by Targeting GFRA4 in Hirschsprung's Disease

DNA and Cell Biology
Gang WangXiangyu Wu

Abstract

Recent studies have suggested the critical roles of miRNAs for disease progression. miRNA-483-5p (miR-483-5p) was previously found to have a relationship with tumor cell behavior, but its biological function in Hirschsprung's disease (HSCR) remains undefined. Thus, we explored the role of miR-483-5p in the pathogenesis of HSCR. Histological changes of colonic tissues were evaluated by hematoxylin and eosin (HE) staining. Quantitative real-time PCR and western blotting were used to determine relative expression levels of miRNA, mRNA, and proteins in 20 HSCR patients and 20 normal colon tissues. In this study, we found that miR-483-5p expression in HSCR tissues was significantly increased and their downregulation promoted cell proliferation, cell cycle progression and invasion and inhibited cell apoptosis in human 293T and SH-SY5Y cell lines by the CCK-8, flow cytometry, and Transwell assay. GNDF family receptor alpha 4 (GFRA4) was confirmed as a downstream target of miR-483-5p by dual-luciferase reporter gene assay and inversely correlated with miR-483-5p expression in cell lines. Taken together, miR-483-5p may play a crucial role in the pathogenesis of HSCR by targeting GFRA4.

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Citations

Jun 30, 2019·International Journal of Molecular Sciences·Ana TorroglosaSalud Borrego
Aug 28, 2020·Pharmacogenomics and Personalized Medicine·Yun ZhuWei Zhong
Feb 23, 2021·Molecular Therapy Oncolytics·Vijay G BhojDon L Siegel

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Methods Mentioned

BETA
protein assay
electrophoresis
transfection
Flow Cytometry
Assay
PCR

Software Mentioned

TargetScan

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