title: Hepatocyte growth factor induces colonic cancer cell invasiveness via enhanced motility and protease overproduction. Evidence for PI3 kinase and PKC involvement.
abstract: Tumour progression to the metastatic phenotype is mainly dependent on tumour cell invasiveness. Cell migration is a crucial step in this process. Here we investigate the effect of hepatocyte growth factor (HGF) on the induction of in vitro invasiveness of poorly aggressive Caco-2 colonic cancer epithelial cells. Invasion assays through a Matrigel barrier were performed. Proteases were assessed by zymography, reverse transcription-polymerase chain reaction and immunoblotting. Caco-2 cells were found to express HGF receptor but not HGF and to secrete several proteases, namely matrix metalloproteinase-1 (MMP-1), MMP-2, possibly MMP-9 and urokinase plasminogen activator (uPA). Exogenous HGF promoted invasiveness of Caco-2 cells through an artificial basement membrane matrix and enhanced their production of proteases. In addition, analyses of media at the end of invasion assays indicated that anti-HGF antibody inhibited protease production in parallel with cell invasion. The involvement of proteases in the HGF-induced invasion process was further investigated using either a synthetic general MMP inhibitor or neutralizing antibodies against MMPs or uPA. All components significantly inhibited HGF-promoted cell invasion. Moreover, specific inhibitors of PKCalpha/beta1 and PI3 kinase also decreased both HGF-promoted cell invasion and protease expression in invasion assay media. Thus, our findings provide evidence that the process of HGF-activated invasiveness of Caco-2 cells involves PI3 kinase and PKC and results from close association of two events, stimulation of cell motile activity and concomitant overproduction of proteases, which permits cell migration through a degraded extracellular matrix.
authors.affiliation: Unités INSERM U 10 and U 410, IFR Cellules Epithéliales, Faculté de Médecine Xavier Bichat, 16 Rue Henri Huchard, BP 416, 75870 Paris, Cedex 18, France.
authors.forename: S
authors.initials: S
authors.lastname: Kermorgant
authors.forename: T
authors.initials: T
authors.lastname: Aparicio
authors.forename: V
authors.initials: V
authors.lastname: Dessirier
authors.forename: M J
authors.initials: MJ
authors.lastname: Lewin
authors.forename: T
authors.initials: T
authors.lastname: Lehy
mesh_terms: D018938:Caco-2 Cells; D003110:Colonic Neoplasms / Q000201:enzymology / Q000473:pathology*; D017228:Hepatocyte Growth Factor / Q000502:physiology*; D006801:Humans; D020782:Matrix Metalloproteinases / Q000096:biosynthesis / Q000378:metabolism*; D009361:Neoplasm Invasiveness; D009362:Neoplasm Metastasis; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D011493:Protein Kinase C / Q000378:metabolism*; D019859:Proto-Oncogene Proteins c-met / Q000378:metabolism
chemical_list: D017228:Hepatocyte Growth Factor; D019869:Phosphatidylinositol 3-Kinases; D019859:Proto-Oncogene Proteins c-met; D011493:Protein Kinase C; D020782:Matrix Metalloproteinases