Downregulation of microRNA-145 may contribute to liver fibrosis in biliary atresia by targeting ADD3

Abstract

Background and objectivesBiliary atresia (BA) is a pediatric liver disease characterized by fibro-obliteration and obstruction of the extrahepatic biliary system, that invariably leads to cirrhosis and even death, if left untreated for extended time. However, its pathology and etiology still remained unknown. In this study, we tested the expression of adducin 3 (ADD3), the gene identified as a susceptibility gene in BA by GWAS, and uncovered its upstream regulatory microRNA in the pathogenesis of BA.MethodsIn this study, 14 infants with BA and 14 infants with choledochal cyst (CC) were enrolled as experimental group and control group, respectively. ADD3 and microRNA-145 (miR-145) expression profiles in liver tissues of BA and CC were determined using qPCR. Luciferase reporter assay was performed to verify the direct interaction between miR-145-5p and ADD3 3’ Untranslated Regions (3’UTR). The Lentiviral vectors containing miR-145, miR-145-3p inhibitor, miR-145-5p inhibitor, empty vector were transfected into human hepatic stellate cell line (LX-2) to determine the functional effect of miR-145 on ADD3 expression at both mRNA and protein level.ResultsMiR-145 was shown to be down-regulated in liver tissues of infants with BA compared to CC (p = 0.0267). ADD3, verified as a target of miR-145-5p, was shown to be overexpressed in infants with BA at the mRNA level (p = 0.0118). Transfection of lentiviruses containing miR-145 into LX-2 cells decreased the expression of ADD3 at both mRNA and protein level compared to negative control group, and suppressed the expression of p-Akt at protein level.ConclusionsOur study has shown that overexpressed ADD3 and downregulated miR-145 were detected in BA liver tissues. MiR-145-5p was confirmed to target ADD3 by luciferase reporter assay. The downregulation of miR-145 may contribute to liver fibrosis in BA by upregulating the expression of ADD3.

A Review on Adducin from Functional to Pathological Mechanisms: Future Direction in Cancer

“…A study using zebrafish model has demonstrated that the loss of add3a, but not xpnpep1 , could lead to impaired biliary function and intrahepatic defects possibly through suppressing hedgehog pathways [ 151 ]. In addition, upregulated expression of ADD3 is also reported to contribute in liver fibrosis in BA and associated with downregulation of miR-145 which targets ADD3 [ 152 ].…”

let-7a suppresses liver fibrosis via TGFβ/SMAD signaling transduction pathway

“…When HSCs are activated, they become fibrogenic myofibroblasts and exhibit the function of fibrogenic myofibroblast-like cells, which can secrete α-smooth muscle actin (α-SMA), TIMP-1 and inhibit HSC cell apoptosis and collagen-I secretion (13–15). Excessive insoluble collagen I and matrix components in the intracellular and perisinusoidal spaces can bring severe compromise to the hepatic function (16,17).…”

Cocaine addicted to cytoskeletal change and a fibrosis high

“…Further to these functions, recent literature has highlighted the role of adducins in T‐cell activation and stimulation of migration, with immunofluorescence techniques demonstrating migratory‐dependent localisation of adducins in CD4+ T‐cells (Butte & Thauland, ). A study by Ye et al () demonstrated the role of miRNA‐145, a regulatory miRNA, in stimulating HSC activation and proliferation via the Wnt/β‐catenin pathway and adducin‐3 (Zhou et al, ). The authors reported the differential expression of miRNA‐145 and adducin‐3 in hepatic tissues of biliary atresia samples.…”

“…Further to these functions, recent literature has highlighted the role of adducins in T-cell activation and stimulation of migration, with immunofluorescence techniques demonstrating migratory-dependent localisation of adducins in CD4+ T-cells (Butte & Thauland, 2015). A study by Ye et al (2017)…”