RhoA promotes differentiation of WB-F344 cells into the biliary lineage.
When cultured on Matrigel, liver precursor epithelium WB-F344 cells could be induced to differentiate into biliary cells in which RhoA expression was upregulated. To further investigate the role of RhoA in WB cell differentiation initiated by Matrigel treatment, we constructed constitutively active RhoA-expressing vectors and stably transfected them into WB-F344 cells. Accompanying upregulation of biliary lineage markers and morphological changes, cells with ectopically active RhoA expression were found to form bile-duct-like structures even without Matrigel treatment. Besides, ROCK inhibitor Y27632 treatment eliminated luminal morphogenesis. F-actin cytoplasmic staining further verified that the RhoA-ROCK signal pathway was involved in differentiation of WB cells into the biliary lineage. In conclusion, our results suggested that the RhoA-ROCK-stress fibre system plays an obligatory role in Matrigel-induced WB-F344 cell luminal morphogenesis and further differentiation.
Yao
Hailei
H
Stem Cells and Regenerative Medicine Lab, Beijing Institute of Transfusion Medicine, Beijing 100850, China.
Jia
Yali
Y
Zhou
Junnian
J
Wang
Jing
J
Li
Yanhua
Y
Wang
Yunfang
Y
Yue
Wen
W
Pei
Xuetao
X
D000577:Amides / Q000494:pharmacology; D000818:Animals; D001483:Base Sequence; D001652:Bile Ducts / Q000254:growth & development; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D019070:Cell Lineage; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004847:Epithelial Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D005822:Genetic Vectors; D008099:Liver / Q000166:cytology*; D008969:Molecular Sequence Data; D011725:Pyridines / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D015854:Up-Regulation; D020742:rhoA GTP-Binding Protein / Q000235:genetics / Q000378:metabolism*
D000577:Amides; D004791:Enzyme Inhibitors; D011725:Pyridines; D012333:RNA, Messenger; C108830:Y 27632; D020742:rhoA GTP-Binding Protein