Hepatocyte growth factor-induced proliferation of hepatic stem-like cells depends on activation of NF-kappaB

Hepatocyte growth factor (HGF) regulates proliferation of hepatic stem cells. Transcription factor nuclear factor kappa B (NF-kappaB) has been demonstrated as a key mediator for cell growth regulation. We investigated the role of NF-kappaB in HGF-mediated cellular proliferation responses in a rat liver-derived hepatic stem-like cell line WB-F344. Cell proliferation was determined by incorporation of [3H]thymidine. Phosphorylation of ERK1/2, p38 MAPK, Akt and IkappaBalpha by HGF stimulation was detected by Western blotting. NF-kappaB activation was determined by electrophoretic mobility shift assay and NF-kappaB-mediated SEAP reporter assay. NF-kappaB activation was inhibited by treatment with an IkappaBalpha dominant-negative vector or inhibitor BAY-11-7082. We found that stimulation of WB-F344 cells with HGF promoted cell proliferation and effectively protected WB-F344 cells from apoptosis induced by TNF-alpha. We also observed activation of ERK1/2, p38 MAPK, Akt and NF-kappaB signaling pathways by HGF in WB-F344 cells. HGF-induced cell proliferation was partly blocked by pre-treatment of the cells with inhibitors against MEK1 or p38 MAPK, and completely blocked using an inhibitor for NF-kappaB activity. Furthermore, it was demonstrated that IkappaB mutant that suppressed NF-kappaB activity completely blocked HGF-induced cell proliferation. NF-kappaB activity is required for HGF-induced proliferation in hepatic stem-like cell line WB-F344, and this activity requires ERK1/2 and p38 MAPK pathways.