Autocrine motility factor enhances hepatoma cell invasion across the basement membrane through activation of beta1 integrins.
Autocrine motility factor/phosphohexose isomerase (AMF/PHI) is a cytokine that is linked to tumor invasion and metastasis. In hepatocellular carcinoma (HCC) tissues, hepatoma cells produce AMF/PHI and its receptor, Mr 78,000 glycoprotein (gp78), is strongly detected in hepatoma cells invading into the stroma and tumor thrombi in the portal vein. Here, we investigated the mechanism of hepatoma cell invasion through Matrigel induced by AMF/PHI using 3 hepatoma cell lines. Production of AMF/PHI, phosphorylation of MEK1/2, and Rho activity were investigated by immunoblotting. Expression of AMF/PHI and gp78 was observed by confocal fluorescence microscopy. The influence of AMF/PHI on activated integrin beta1 subunit expression was evaluated by flow cytometry. Changes in invasion, adhesion, and motility induced by AMF/PHI were evaluated using chemoinvasion, adhesion, and phagokinetic track motility assays. The effect of AMF/PHI on matrix metalloproteinase (MMP) secretion was evaluated by gelatin zymography. Hepatoma cells produced AMF/PHI and expressed gp78. Although AMF/PHI was ubiquitously detected, gp78 was strongly expressed in migrating cells. AMF/PHI induced up-regulation of activated integrin beta1 subunit expression. AMF/PHI stimulated hepatoma cell invasion through Matrigel, and stimulated the adhesion, motility, and MMP-2 secretion of hepatoma cells. The latter effects were suppressed by the function-blocking antibody for integrin beta1 subunit. AMF/PHI also enhanced Rho activity and the phosphorylation of MEK1 and MEK 2. Our results indicate that AMF/PHI enhances hepatoma cell invasion through Matrigel in an autocrine manner by stimulating the adhesion, motility, and MMP-2 secretion of these cells through activation of beta1 integrins.
Torimura
T
T
The Second Department of Medicine and Research Center for Innovative Cancer Therapy, Kurume University School of Medicine, Kurume City, Japan. tori@med.kurume-u.ac.jp
Ueno
T
T
Kin
M
M
Harada
R
R
Nakamura
T
T
Kawaguchi
T
T
Harada
M
M
Kumashiro
R
R
Watanabe
H
H
Avraham
R
R
Sata
M
M
D001485:Basement Membrane / Q000473:pathology*; D006528:Carcinoma, Hepatocellular / Q000737:chemistry / Q000378:metabolism / Q000473:pathology*; D002448:Cell Adhesion; D003094:Collagen; D004338:Drug Combinations; D004591:Electrophoresis, Polyacrylamide Gel; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D005956:Glucose-6-Phosphate Isomerase / Q000032:analysis / Q000096:biosynthesis / Q000494:pharmacology*; D006801:Humans; D015151:Immunoblotting; D019012:Integrin beta1 / Q000032:analysis / Q000502:physiology*; D007797:Laminin; D008113:Liver Neoplasms / Q000737:chemistry / Q000378:metabolism / Q000473:pathology*; D048369:MAP Kinase Kinase 1; D048370:MAP Kinase Kinase 2; D020782:Matrix Metalloproteinases / Q000096:biosynthesis; D020929:Mitogen-Activated Protein Kinase Kinases / Q000378:metabolism; D009361:Neoplasm Invasiveness; D010766:Phosphorylation; D011169:Portal Vein / Q000473:pathology; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D011509:Proteoglycans; D060285:Receptors, Autocrine Motility Factor; D018121:Receptors, Cytokine / Q000032:analysis; D014407:Tumor Cells, Cultured; D044767:Ubiquitin-Protein Ligases
D004338:Drug Combinations; D019012:Integrin beta1; D007797:Laminin; D011509:Proteoglycans; D018121:Receptors, Cytokine; C059485:matrigel; D003094:Collagen; C555000:AMFR protein, human; D060285:Receptors, Autocrine Motility Factor; D044767:Ubiquitin-Protein Ligases; C482128:MAP2K2 protein, human; D011505:Protein-Tyrosine Kinases; D017346:Protein Serine-Threonine Kinases; D048369:MAP Kinase Kinase 1; D048370:MAP Kinase Kinase 2; C482125:MAP2K1 protein, human; D020929:Mitogen-Activated Protein Kinase Kinases; D020782:Matrix Metalloproteinases; D005956:Glucose-6-Phosphate Isomerase