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

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.

Keywords

• atpase family aaa domain protein
• normal rat liver stem cells
• epithelial-to-mesenchymal transition
• invasion
• migration