Identification of metastasis-promoting sequences in the mouse laminin alpha 1 chain.
Laminin-1, a major basement membrane matrix glycoprotein, enhances adhesion, migration, and metastasis of tumor cells. We have screened 208 overlapping synthetic peptides covering the short and long arms of mouse laminin alpha1 chain for their adhesion activity with B16-F10 mouse melanoma cells. Cell adhesion activity was determined using various amounts of peptides coated on plastic dishes and by measuring cell adhesion on peptide-conjugated Sepharose beads. Nineteen peptides showed B16-F10 cell adhesion activity. Three peptides, designated A-13, -24, and -208, showed the strongest attachment activity in the plate assay, whereas 4 peptides, A-13, -51, -99, and -112, demonstrated the strongest cell adhesion when conjugated to beads. The 19 peptides were tested in vivo for their effect on experimental pulmonary metastasis by B16-F10 cells. Four peptides, A-13, -51, -64, and -119, significantly enhanced metastasis, with A-13 showing the strongest dramatic enhancement. The four metastasis-promoting peptides also stimulated migration of B16-F10 cells in the Boyden chamber assay in vitro with A-13 being the most potent stimulator. In addition, the 4 peptides inhibited laminin-induced cell attachment and migration, which indicates that these four sequences are possible functional B16-F10 cell binding sites in laminin-1. All the four sequences are located on the globular domains of the short arm. Other peptides, including strong adhesion-active peptides, A-24, -99, -112, and a scrambled A-13 peptide, did not stimulate either migration or metastasis. Thus, laminin-1 has multiple active sites in the globular domains of the short arm which promote migration and metastasis of B16-F10 cells.
Kuratomi
Y
Y
National Institute of Dental and Craniofacial Research, NIH, Bethesda, Maryland, 20892, USA.
Nomizu
M
M
Nielsen
P K
PK
Tanaka
K
K
Song
S Y
SY
Kleinman
H K
HK
Yamada
Y
Y
D000818:Animals; D002448:Cell Adhesion; D002465:Cell Movement; D007275:Injections, Intravenous; D007797:Laminin / Q000737:chemistry / Q000502:physiology*; D008175:Lung Neoplasms / Q000473:pathology / Q000556:secondary*; D008546:Melanoma, Experimental / Q000378:metabolism / Q000473:pathology / Q000556:secondary*; D051379:Mice; D008810:Mice, Inbred C57BL; D008863:Microspheres; D009368:Neoplasm Transplantation; D010455:Peptides / Q000378:metabolism / Q000502:physiology*
D007797:Laminin; D010455:Peptides; C084098:laminin A