Mitogenic activity toward human breast cancer cell line MCF-7 of two bFGFs purified from sera of breast cancer patients: co-operative role of cathepsin D.
Mitogenic activity toward MCF-7 cells of two immunoreactive (high-molecular-weight form bFGF, HMW-bFGF; and 16-K bFGF, having the same molecular weight as recombinant bFGF) purified from pooled sera of breast cancer patients by heparin-affinity chromatography and gel filtration was investigated. The mitogenic activity of 16-K bFGF toward the cells was equal to that of recombinant bFGF, whereas the mitogenic effect of HMW-bFGF was weak. Most of the mitogenic activity of these two bFGFs was neutralized by anti-bFGF antibody. Also, the mitogenic activity of both HMW-bFGF and 16-K bFGF was markedly enhanced by aspartyl protease (cathepsin D), which is secreted in excess by breast cancer cells and is responsible for the enzymatic degradation of the extracellular matrix (ECM). By an enzyme immunoassay, we detected cathepsin D-mediated release of recombinant bFGF previously bound to the ECM of MCF-7 cells into the conditioned medium, and also observed cathepsin D-mediated proteolysis of HMW-bFGF to release free 16-K bFGF. These results suggest that 16-K bFGF is the bFGF molecule itself in the blood and that HMW-bFGF is a circulating form of bFGF in blood whose mitogenic activity is regulated by cathepsin D.
Takei
Y
Y
Department of Molecular Biology, Gifu Pharmaceutical University, Japan.
Higashira
H
H
Yamamoto
T
T
Hayashi
K
K
D000906:Antibodies / Q000276:immunology; D001943:Breast Neoplasms / Q000097:blood / Q000473:pathology*; D002402:Cathepsin D / Q000494:pharmacology; D002455:Cell Division / Q000187:drug effects*; D017077:Culture Media, Conditioned; D005260:Female; D016222:Fibroblast Growth Factor 2 / Q000276:immunology / Q000302:isolation & purification* / Q000494:pharmacology*; D006801:Humans; D008970:Molecular Weight; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000473:pathology
D000906:Antibodies; D017077:Culture Media, Conditioned; D016222:Fibroblast Growth Factor 2; D002402:Cathepsin D