Mechanism of the transforming growth factor-beta induction of fibronectin expression in hepatic stem-like cells.

Transforming growth factor-beta1 (TGF-beta1) plays an important role in the fibrogenic process in the liver. The aim of the present study was to explore the action of TGF-beta1 on fibronectin expression in rat hepatic stem-like cells and the underlying mechanisms. The level of fibronectin expression was determined in hepatic stem-like cells (WB cells) before and after TGF-beta1 stimulation by RT-PCR and Western blot methods. Using immunogold transmission electron microscopy and the Western blot method, we observed the result of the expression and the distribution of cAMP, phosphorylated Smad3 and Smad7 before and after TGF-beta1 treatment. The levels of fibronectin expression in both mRNA and protein increased 4- to 5-fold after TGF-beta1 stimulation, reaching an optimum level after 8 h and then gradually falling back. Similarly, TGF-beta1 stimulation resulted in an increase of cAMP in WB cells, peaking at 8 h. After treatment with TGF-beta1 for 24 h, the expression of cAMP gradually decreased. In addition, we found that TGF-beta1 treatment also contributed to the increased expression and to changes in cellular distribution of phosphorylated Smad3 (translocation from the cytoplasm to the nucleus) and Smad7 (translocation from the nucleus to the cytoplasm) in WB cells. The present study demonstrates that TGF-beta is involved in the fibrogenic process in hepatic stem cells through up-regulation of fibronectin expression, and the mechanisms underlying this process may be associated with the activation of cAMP and Smad pathways.