Regulation of melanoma cell migration and invasion by laminin-5 and alpha3beta1 integrin (VLA-3).
We evaluated the role of soluble factors produced from epidermal cells in melanoma cell motility by using the Boyden chamber chemoinvasion system. The migration of two melanoma cell lines, A375 and Mewo, was potentiated by conditioned media of A431 epidermoid cells in a concentration-dependent manner. The enhancement of A375 melanoma cell motility induced by the conditioned medium was blocked by antibodies against either alpha3 or beta1 integrin subunit. The motility-stimulating activity was recovered in the same fraction as the alpha3 integrin-dependent adhesion-promoting activity in a high-molecular-weight (>200 kDa) fraction on Superose 12 gel chromatography, and adsorbed with an anti-laminin-5 antibody. Purified laminin-5 was capable of potentiating melanoma cell migration as measured in either the chemotaxis assay with a soluble form of laminin-5 or the haptotaxis assay with membranes coated with a mixture of laminin-5 and Matrigel. Furthermore, immobilized laminin-5 induced A375 melanoma cells to secrete matrix metalloproteinase-9 (type IV collagenase) into the culture medium. These results strongly suggest that the interaction of laminin-5 produced in the epidermis with alpha3beta1 integrin on melanoma cells is involved in cell migration, invasion, and degradation of extracellular matrix proteins.
Tsuji
Tsutomu
T
Department of Microbiology, Hoshi University, Tokyo, Japan. tsuji@hoshi.ac.jp
Kawada
Yoko
Y
Kai-Murozono
Mieko
M
Komatsu
Shinya
S
Han
Seon Ae
SA
Takeuchi
Ken-ichi
K
Mizushima
Hiroto
H
Miyazaki
Kaoru
K
Irimura
Tatsuro
T
D015815:Cell Adhesion Molecules / Q000378:metabolism* / Q000494:pharmacology; D002465:Cell Movement / Q000502:physiology*; D003094:Collagen / Q000737:chemistry; D017077:Culture Media, Conditioned; D004338:Drug Combinations; D005434:Flow Cytometry; D005780:Gelatin / Q000378:metabolism; D006801:Humans; D039201:Integrin alpha3beta1; D016023:Integrins / Q000378:metabolism*; D007797:Laminin / Q000737:chemistry; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D008545:Melanoma / Q000378:metabolism* / Q000473:pathology; D009361:Neoplasm Invasiveness; D011509:Proteoglycans / Q000737:chemistry; D011994:Recombinant Proteins / Q000378:metabolism; D012878:Skin Neoplasms / Q000378:metabolism* / Q000473:pathology; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000473:pathology; D000097794:Kalinin
D015815:Cell Adhesion Molecules; D003094:Collagen; D017077:Culture Media, Conditioned; D004338:Drug Combinations; D005780:Gelatin; D039201:Integrin alpha3beta1; D016023:Integrins; D007797:Laminin; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D011509:Proteoglycans; D011994:Recombinant Proteins; D000097794:Kalinin; C059485:matrigel