title: Possible involvement of p21/waf1 in the growth inhibition of HepG2 cells induced by hepatocyte growth factor.
abstract: Hepatocyte growth factor (HGF) is a potent mitogen for a variety of cell types, but it is also known as an antimitogenic factor for several types of tumor cell lines. The biological processes by which HGF inhibits tumor cell growth remain poorly understood. Here we report a comparative study of HGF-mediated signal transduction events between two opposite responding types of human hepatoblastoma cell lines, HuH6 and HepG2. Following serum starvation, both cell lines were cultured in hepatocyte growth medium (HGM), a chemically defined medium, in the presence or absence of HGF. Under these culture conditions, cell growth in HuH6 was promoted by HGF, while it was inhibited in HepG2. Phosphorylation of p42/mitogen-activated protein (MAP) kinase was observed within 10 min after HGF stimulation in both cell lines. The level of phosphorylated MAP kinase in HuH6 declined to basal levels after 2 hr. However, in HepG2 the phosphorylated form was detectable at 6 hr. p21/waf1 was induced in both cell lines where levels peaked 4-6 hr after HGF stimulation. In HuH6, a marked decrease of p21/waf1 was observed at 8-12 hr, while a high level of p21/waf1 was sustained for at least 24 hr in HepG2. HGF treatment depressed cdk2 activity in a time-dependent manner in HepG2 while the activity increased in HuH6. When serum-starved HepG2 was growth stimulated with serum in the presence or absence of HGF, the cells treated with HGF underwent growth inhibition correlating with a sustained induction of p21/waf1 and a decrease of cdk2 activity. Immunoprecipitation analysis revealed accumulation of cdk2-associated p21/waf1 in the HGF-treated HepG2. Together, the results suggest that sustained induction of p21/waf1 mediates growth inhibition in HepG2 in the presence of HGF.
authors.affiliation: Research Institute of Life Science, Snow Brand Milk Products, Co., Ltd., Tochigi, Japan. fvbd7042@mb.infoweb.or.jp
authors.forename: N
authors.initials: N
authors.lastname: Shima
authors.forename: D B
authors.initials: DB
authors.lastname: Stolz
authors.forename: M
authors.initials: M
authors.lastname: Miyazaki
authors.forename: E
authors.initials: E
authors.lastname: Gohda
authors.forename: K
authors.initials: K
authors.lastname: Higashio
authors.forename: G K
authors.initials: GK
authors.lastname: Michalopoulos
mesh_terms: D001798:Blood Proteins / Q000494:pharmacology; D042846:CDC2-CDC28 Kinases; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D051357:Cyclin-Dependent Kinase 2; D051358:Cyclin-Dependent Kinase 4; D050759:Cyclin-Dependent Kinase Inhibitor p21; D018844:Cyclin-Dependent Kinases / Q000096:biosynthesis / Q000378:metabolism; D016213:Cyclins / Q000096:biosynthesis / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000378:metabolism*; D018197:Hepatoblastoma; D017228:Hepatocyte Growth Factor / Q000494:pharmacology*; D006801:Humans; D007700:Kinetics; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000096:biosynthesis / Q000378:metabolism; D011518:Proto-Oncogene Proteins; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D014407:Tumor Cells, Cultured / Q000166:cytology / Q000187:drug effects / Q000201:enzymology
chemical_list: D001798:Blood Proteins; C493956:CDKN1A protein, human; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D004791:Enzyme Inhibitors; D011518:Proto-Oncogene Proteins; D017228:Hepatocyte Growth Factor; D017346:Protein Serine-Threonine Kinases; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D042846:CDC2-CDC28 Kinases; C495894:CDK2 protein, human; C495900:CDK4 protein, human; D051357:Cyclin-Dependent Kinase 2; D051358:Cyclin-Dependent Kinase 4; D018844:Cyclin-Dependent Kinases