Overexpression of ATPase family AAA domain containing protein 3A induces epithelial-to-mesenchymal transition of normal rat liver stem cells <italic style="font-style: italic">in vitro</italic>

Yaxing FENG; Longfu ZHOU; Yihan WANG; Rui GU; Wei LI; Yaolei ZHANG; Ting LI; Weihui LIU

doi:10.16016/j.1000-5404.201907114

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Overexpression of ATPase family AAA domain containing protein 3A induces epithelial-to-mesenchymal transition of normal rat liver stem cells in vitro

Basic Medicine | 更新时间：2026-01-09

- Overexpression of ATPase family AAA domain containing protein 3A induces epithelial-to-mesenchymal transition of normal rat liver stem cells in vitro
- Journal of Third Military Medical University Vol. 41, Issue 24, Pages: 2417-2424(2019)
- 作者机构：
  
  1.610003 成都，西南交通大学医学院
  2.610083 成都，中国人民解放军西部战区总医院中心实验室
- 作者简介：
- 基金信息：
  
  the Key Applied Science Research Project of Sichuan Provincial Department of Science and Technology(2019YJ0586)
- DOI：10.16016/j.1000-5404.201907114
  CLC： R394.3;R730.23;R735.7
- Received：15 July 2019，
  
  Revised：2019-09-19，
  
  Published：30 December 2019
- 稿件说明：
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Yaxing FENG, Longfu ZHOU, Yihan WANG, et al. Overexpression of ATPase family AAA domain containing protein 3A induces epithelial-to-mesenchymal transition of normal rat liver stem cells in vitro[J]. Journal of Third Military Medical University, 2019, 41(24): 2417-2424.
DOI：

Yaxing FENG, Longfu ZHOU, Yihan WANG, et al. Overexpression of ATPase family AAA domain containing protein 3A induces epithelial-to-mesenchymal transition of normal rat liver stem cells in vitro[J]. Journal of Third Military Medical University, 2019, 41(24): 2417-2424. DOI： 10.16016/j.1000-5404.201907114.

摘要

目的

探讨过表达ATP酶家族AAA结构域蛋白(ATPase family AAA domain protein，ATAD3A)后对大鼠肝脏干细胞上皮-间质转化(epithelial-to-mesenchymal transition，EMT)、迁移和侵袭能力的影响。

方法

体外培养大鼠肝干细胞系(WBF-344)，分别转染含过表达ATAD3A基因的慢病毒(ATAD3A过表达组)和不含过表达ATAD3A基因的载体(空载对照组)，以不做处理的细胞作为正常对照组。显微镜下观察细胞形态变化，细胞划痕和Transwell实验检测各组细胞的迁移和侵袭能力，RT-qPCR法和Western blot法检测EMT标志物和转录因子的基因和蛋白表达水平，体外裸鼠皮下成瘤实验验证肝干细胞恶性转化程度。

结果

与2个对照组比较，ATAD3A过表达组:①细胞由卵圆形向长条形和梭形的间质细胞形态特征转变；②细胞迁移和侵袭能力得到明显增强(

0.05)；③E-cadherin的基因和蛋白表达明显降低，而N-cadherin的基因和蛋白表达明显升高(

0.05)；④转录因子ZEB1和ZEB2的表达也显著增加(

0.05)。但体外成瘤实验未能观察到肿瘤块的形成。

结论

过表达ATAD3A后能诱导大鼠肝干细胞EMT的发生。

Abstract

Objective

To investigate the effect of overexpression of ATPase family AAA domain protein 3A (ATAD3A) on epithelial-to-mesenchymal transition (EMT)

migration and invasion of rat liver stem cells.

Methods

Cultured rat liver stem cell line (WBF-344) was transfected with a lentivirus vector carrying ATAD3A to construct a stable ATAD3A-overexpressing cell line. The morphological changes of the transfected cells were observed microscopically

and the migration and invasion ability of the cells were assessed using cell scratch and Transwell assays. The expression of the EMT markers and transcription factors at the mRNA and protein levels was detected using RT-qPCR and Western blotting. The malignant transformation of the transfected cells was evaluated by assessing their subcutaneous tumorigenic ability in nude mice.

Results

Compared with the cells transfected with the blank lentiviral vector

the cells overexpressing ATAD3A showed obvious morphological transformation from an oval to an elongated and spindle-shaped morphology and exhibited significantly enhanced migration and invasion abilities. ATAD3A overexpression in WBF-344 cells significantly decreased the expression of E-cadherin and increased the expression of N-cadherin at both the mRNA and protein levels

causing also significant enhancement of ZEB1 and ZEB2 expression. In the

in vivo

experiment

subcutaneous implantation of the transfected cells did not result in tumor formation in nude mice.

Conclusion

Overexpression of ATAD3A can induce EMT in normal rat liver stem cells.

关键词

Keywords

references

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Overexpression of ATPase family AAA domain containing protein 3A induces epithelial-to-mesenchymal transition of normal rat liver stem cells in vitro

DOI：10.16016/j.1000-5404.201907114

摘要

Abstract

关键词

Keywords

references