Overexpression of miR-122 promotes the hepatic differentiation and maturation of mouse ESCs through a miR-122/FoxA1/HNF4a-positive feedback loop

被引：110

作者：

Deng, Xiao-Geng ^{[1
]}

Qiu, Rong-Lin ^{[1
]}

Wu, Yao-Hao ^{[1
]}

Li, Zhi-Xi ^{[1
]}

Xie, Ping ^{[1
]}

Zhang, Jie ^{[1
]}

Zhou, Jia-Jia ^{[1
]}

Zeng, Le-Xiang ^{[1
]}

Tang, Jing ^{[1
]}

Maharjan, Anu ^{[1
]}

Deng, Jie-Min ^{[2
]}

机构：

[1] Sun Yat Sen Univ, Mem Hosp, Dept Pediat Surg, Guangzhou 510120, Peoples R China

[2] Sun Yat Sen Univ, Guangdong Higher Educ Inst, Key Lab Malignant Tumor Gene Regulat & Target The, Guangzhou 510120, Peoples R China

来源：

LIVER INTERNATIONAL | 2014年 / 34卷 / 02期

基金：

中国国家自然科学基金;

关键词：

differentiation; embryonic stem cells; hepatocytes; mesenchymal-to-epithelial transition; miR-122; EMBRYONIC STEM-CELLS; HEPATOCYTE-LIKE CELLS; HEPATOCELLULAR-CARCINOMA; LIVER DEVELOPMENT; EXPRESSION; ACTIVATION; IDENTIFICATION; RECRUITMENT; ENDODERM; PROTEIN;

D O I：

10.1111/liv.12239

中图分类号：

R57 [消化系及腹部疾病];

学科分类号：

100201 [内科学];

摘要：

Background & AimsmicroRNA-122 is the only identified liver-specific miRNA and plays a crucial role in liver development, maintenance of hepatic homeostasis as well as tumourigenesis. In our previous differentiation of ESCs into hepatocytes, microRNA-122 (miR-122) was expressed at a relatively low level. Here, we aim to elucidate the effect and underlying mechanisms of miR-122 during differentiation of ESCs into hepatocytes. MethodsMouse ESCs were initially induced towards HPCs by activin A, FGF-4 and sodium butyrate and were subsequently transfected with a recombinant adenovirus expressing vector pAV.Ex1d-CMV>miR-122/IRES/eGFP 9days after induction. Cells were analysed by real-time PCR, immunofluorescence, flow cytometry, microscopy and functional assays. Furthermore, microarray analysis was performed. ResultsWe demonstrated that overexpression of miR-122 could effectively promote hepatic differentiation and maturation, as assessed by morphological and functional tests. The microarray analysis revealed that 323 genes were down-regulated, whereas 59 were up-regulated. Particularly, two liver-specific transcription factors, FoxA1 and HNF4a, were significantly up-regulated. Moreover, the expression of E-cadherin was dramatically increased and the proliferation of HPCs was suppressed, whereas knockdown of FoxA1 reduced E-cadherin expression and increased the proliferation of HPCs. In addition, the expression levels of FoxA1, HNF4a and E-cadherin in time-course transfection experiments with miR-122 were not significantly increased except in cells in which transfection with miR-122 occurred 9days after induction. ConclusionOverexpression of miR-122 at an appropriate stage could promote hepatic differentiation and maturation by regulating the balance between proliferation and differentiation, as well as the balance between EMT and MET, partially through a miR-122/FoxA1/HNF4a-positive feedback loop.

引用

页码：281 / 295

页数：15

共 47 条

[1]

TGF-β inactivation and TGF-α overexpression cooperate in an in vivo mouse model to induce hepatocellular carcinoma that recapitulates molecular features of human liver cancer [J].