Inhibition of cell migration and angiogenesis by the amino-terminal fragment of 24kD basic fibroblast growth factor.
The 24-kDa form of basic fibroblast growth factor inhibits the migration of endothelial cells and mammary carcinoma cells while continuing to promote cell proliferation. This molecule consists of the 18-kDa fibroblast growth factor sequence plus an additional 55 amino acids at the amino-terminal end. Antibody neutralization studies suggested that the inhibition of migration is associated with these 55 amino acids, whereas the promotion of proliferation localizes to the 18-kDa domain. To determine whether 24kD basic fibroblast growth factor could be modified to eliminate its effect on cell proliferation but retain its inhibition of migration, portions of the carboxyl-terminal end of 24kD fibroblast growth factor were deleted, and the products were tested on MCF-7 and endothelial cells. A protein consisting of the 55 amino acids of the amino-terminal end and the first 31 amino acids of 18kD basic fibroblast growth factor (ATE+31) inhibited migration by 80% but did not promote cell growth. Arginine to alanine substitutions within the first 21 amino acids of the carboxyl-terminal end substantially reduced the efficacy of ATE+31, whereas substitutions in the remaining part of the molecule had no effect. Competition binding experiments showed that ATE+31 does not compete with 24kD basic fibroblast growth factor for binding to fibroblast growth factor receptor 1. In an in vivo matrigel plug assay, 150 nm ATE+31 peptide reduced angiogenesis by 80%. These studies demonstrate that the amino-terminal end of 24kD basic fibroblast growth factor is responsible for an activity that inhibits the migration rates of cultured cells as well as the angiogenic response in vivo.
Ding
Lan
L
La Jolla Institute for Molecular Medicine, San Diego, CA 92121, USA.
Doñate
Fernando
F
Parry
Graham C N
GC
Guan
Xiaojun
X
Maher
Pamela
P
Levin
Eugene G
EG
D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D002465:Cell Movement / Q000187:drug effects; D005260:Female; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D006801:Humans; D008970:Molecular Weight; D010446:Peptide Fragments / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D014407:Tumor Cells, Cultured
D020533:Angiogenesis Inhibitors; D010446:Peptide Fragments; D016222:Fibroblast Growth Factor 2