HGF-promoted motility in primary human melanocytes depends on CD44v6 regulated via NF-kappa B, Egr-1, and C/EBP-beta.
The regulation of CD44v6, a variant of the CD44 family of glycosylated adhesion molecules, through hepatocyte growth factor (HGF) has implications for motility in primary human melanocytes. We show that exposure of primary human melanocytes to HGF results in an increase of CD44v6 expression. Immunostaining of melanocytic lesions revealed low cytoplasmic positivity of CD44v6 in some nevi but high membranous expression in primary cutaneous melanomas, and cutaneous and lymph node metastases. HGF-dependent CD44v6 regulation in melanocytes is NF-kappaB dependent because BAY 11-7082, an inhibitor of NF-kappaB activation, but not interference with the mitogen-activated protein kinase or phosphatidylinositol 3-kinase cascade, antagonized HGF-induced CD44v6 expression. NF-kappaB-mediated transcriptional regulation of CD44v6 involves the transcription factors Egr-1 and CCAAT enhancer-binding protein-beta (C/EBP-beta). In gel shift assays, the initial binding of p100/p52 NF-kappaB, C/EBP-beta, and Egr-1 to the CD44 promoter experienced reshuffling toward increased affinity of C/EBP-beta after HGF stimulation. A blocking antibody to CD44v6 decreased HGF-induced c-Met phosphorylation as well as enhanced random- and site-directed migration. Our data show that HGF-induced motility in primary human melanocytes depends on c-Met-CD44v6 interaction, and that HGF-enhanced CD44v6 expression is required for motility and transcriptional upregulation of CD44v6, presumably mediated through a complex comprising NF-kappaB/C/EBP-beta and Egr-1.
Damm
Sabine
S
Cancer Biology Unit, Department of Dermatology, Medical University Graz, Auenbruggerplatz 8, Graz, Austria.
Koefinger
Petra
P
Stefan
Martina
M
Wels
Christian
C
Mehes
Gabor
G
Richtig
Erika
E
Kerl
Helmut
H
Otte
Marcus
M
Schaider
Helmut
H
D022782:CCAAT-Enhancer-Binding Protein-beta / Q000378:metabolism; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology*; D051766:Early Growth Response Protein 1 / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic / Q000502:physiology; D017228:Hepatocyte Growth Factor / Q000378:metabolism / Q000494:pharmacology; D006801:Humans; D018960:Hyaluronan Receptors / Q000235:genetics / Q000378:metabolism*; D007150:Immunohistochemistry; D008207:Lymphatic Metastasis; D008544:Melanocytes / Q000378:metabolism / Q000473:pathology*; D008545:Melanoma / Q000378:metabolism / Q000503:physiopathology / Q000556:secondary*; D016328:NF-kappa B / Q000378:metabolism; D009506:Nevus / Q000378:metabolism / Q000473:pathology; D015398:Signal Transduction / Q000502:physiology; D012878:Skin Neoplasms / Q000378:metabolism / Q000473:pathology* / Q000503:physiopathology; D014158:Transcription, Genetic / Q000502:physiology
D022782:CCAAT-Enhancer-Binding Protein-beta; C112697:CD44v6 antigen; C497325:EGR1 protein, human; D051766:Early Growth Response Protein 1; D018960:Hyaluronan Receptors; D016328:NF-kappa B; D017228:Hepatocyte Growth Factor