Growth and differentiation factors inhibit the migratory phenotype of cultured neonatal rat hepatocytes induced by HGF/SF.
Previous studies have demonstrated that neonatal rat hepatocytes cultured in a serum-free medium adopt a fibroblast-like morphology and form a well-developed vimentin network. By immunoblotting, phase-contrast microscopy, and immunolocalization studies we report here that hepatocyte growth factor (HGF) induces a pronounced migratory/scatter phenotype in neonatal hepatocytes in primary culture. These fibroblast-like cells are highly elongated and adopt a pronounced spindle shape in the presence of this growth factor. Most of them coexpress vimentin and cytokeratin intermediate filaments. Both EGF and TGF-beta1 also induced vimentin expression in cytokeratin-positive cells but this effect was not correlated with a change of epithelial phenotype. Only in DMSO-supplemented cultures was vimentin expression of hepatocytes inhibited and no coexpression was observed in the presence of this factor. Remarkably, the effect induced by HGF was totally inhibited when DMSO, TGF-beta1, or EGF was added to HGF-supplemented cultures. Epithelial sheets of well-defined hepatocytes were observed with these combinations although the complete epithelial morphology was achieved by combining DMSO with a growth factor. Taken together these results suggest that growth and differentiation factors modulate the migratory effect of HGF/SF in hepatocytes.
Pagan
R
R
Departament de Biologia Cellular i, Departament de Bioquímica i Biologia Molecular;, Universitat de Barcelona;, Avgda. Diagonal, 645, Barcelona, Catalonia, 08028, Spain.
Martín
I
I
Llobera
M
M
Vilaró
S
S
D000818:Animals; D000831:Animals, Newborn; D002454:Cell Differentiation / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D002478:Cells, Cultured; D004121:Dimethyl Sulfoxide / Q000494:pharmacology*; D004815:Epidermal Growth Factor / Q000494:pharmacology; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D005455:Fluorescent Antibody Technique; D006133:Growth Substances / Q000494:pharmacology*; D017228:Hepatocyte Growth Factor / Q000494:pharmacology*; D007382:Intermediate Filaments / Q000187:drug effects / Q000502:physiology / Q000648:ultrastructure; D007633:Keratins / Q000096:biosynthesis; D008099:Liver / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D010641:Phenotype; D051381:Rats; D016212:Transforming Growth Factor beta / Q000494:pharmacology; D014746:Vimentin / Q000096:biosynthesis
D006133:Growth Substances; D016212:Transforming Growth Factor beta; D014746:Vimentin; D004815:Epidermal Growth Factor; D017228:Hepatocyte Growth Factor; D007633:Keratins; D004121:Dimethyl Sulfoxide