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2017 ; 12
(9
): e0180896
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Downregulation of microRNA-145 may contribute to liver fibrosis in biliary
atresia by targeting ADD3
#MMPMID28902846
Ye Y
; Li Z
; Feng Q
; Chen Z
; Wu Z
; Wang J
; Ye X
; Zhang D
; Liu L
; Gao W
; Zhang L
; Wang B
PLoS One
2017[]; 12
(9
): e0180896
PMID28902846
show ga
BACKGROUND AND OBJECTIVES: Biliary 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. METHODS: In 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. RESULTS: MiR-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. CONCLUSIONS: Our 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.