title: Integration of Ras subeffector signaling in TGF-beta mediated late stage hepatocarcinogenesis.
abstract: Immortalized p19(ARF) null hepatocytes (MIM) feature a high degree of functional differentiation and are susceptible to transforming growth factor (TGF)-beta driven growth arrest and apoptosis. In contrast, polarized MIM hepatocytes expressing hyperactive Ha-Ras continue proliferation in cooperation with TGF-beta, and adopt an invasive phenotype by executing an epithelial to mesenchymal transition (EMT). In this study, we analyzed the involvement of Ras subeffectors in TGF-beta mediated hepatocellular EMT by employing MIM hepatocytes, which express Ras mutants allowing selective activation of either mitogen-activated protein kinase (MAPK) signaling (V12-S35) or phosphoinositide 3-OH (PI3)3 kinase (PI3K) signaling (V12-C40). We found that MAPK signaling in MIM-S35 hepatocytes was necessary and sufficient to promote resistance to TGF-beta mediated inhibition of proliferation in vitro and in vivo. MIM-S35 hepatocytes showed also PI3K activation during EMT, however, MAPK signaling on its own protected hepatocytes from apoptosis. Yet, MIM-C40 hepatocytes failed to form tumors and required additional MAPK stimulation to overcome TGF-beta mediated growth arrest. In vivo, the collaboration of MAPK signaling and TGF-beta activity drastically accelerated the cell-cycle progression of the hepatocytes, leading to vast tumor formation. From these data we conclude that MAPK is crucial for the cooperation with TGF-beta to regulate the proliferation as well as the survival of hepatocytes during EMT, and causes the fatal increase in hepatocellular tumor progression.
authors.affiliation: Department of Medicine I, Institute of Cancer Research, Medical University of Vienna, Borschke-Gasse 8a, A-1090 Vienna, Austria.
authors.forename: Alexandra N M
authors.initials: AN
authors.lastname: Fischer
authors.forename: Blanca
authors.initials: B
authors.lastname: Herrera
authors.forename: Mario
authors.initials: M
authors.lastname: Mikula
authors.forename: Verena
authors.initials: V
authors.lastname: Proell
authors.forename: Eva
authors.initials: E
authors.lastname: Fuchs
authors.forename: Josef
authors.initials: J
authors.lastname: Gotzmann
authors.forename: Rolf
authors.initials: R
authors.lastname: Schulte-Hermann
authors.forename: Hartmut
authors.initials: H
authors.lastname: Beug
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Mikulits
mesh_terms: D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000378:metabolism*; D002471:Cell Transformation, Neoplastic / Q000378:metabolism; D006801:Humans; D051379:Mice; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D009361:Neoplasm Invasiveness; D015398:Signal Transduction / Q000502:physiology*; D016212:Transforming Growth Factor beta / Q000378:metabolism*; D018631:ras Proteins / Q000378:metabolism*
chemical_list: D016212:Transforming Growth Factor beta; D020928:Mitogen-Activated Protein Kinases; D018631:ras Proteins