A novel recombinant basic fibroblast growth factor and its secretion

Basic fibroblast growth factor (FGF-2) is a pleiotropic mitogen which plays an important role in cell growth, differentiation, migration, and survival in different cells and organ systems. Recently, several clinical applicati ons for FGF-2 gene transfer are being evaluated in wound healing and collat eral artery development to relieve myocardial and peripheral ischemia due t o the ability of FGF-2 to regulate the growth and function of vascular cell s. However, FGF-2 lacks a classical hydrophobic secretion signal peptide, t he FGF-2 chimeras containing various signal sequences have been explored. I n this study, a novel recombinant 4sFGF-2 was constructed by replacing nine residues from the amino-terminus of native FGF-2 (Met1 to Leu9) with eight amino acid residues of signal peptide of FGF-4 (Met1 to Ala8) to better in crease the secretion level of FGF-2. When the recombinant FGF-S gene, clone d into the expression vector with CMV promoter, was expressed in COS-7 cell s, the recombinant 4sFGF-2 was highly secreted into the media. The secreted 4sFGF-2 showed the same biological activity as the native FGF-S in the dos e-response effects on DNA synthesis and cell growth of rat aortic smooth mu scle cells (RASMCs) and NIH3T3 cells. The 4sFGF-2 also was able to activate MAPK as wild FGF-2 in RASMCs. These results indicate that a novel recombin ant 4sFGF-2 may be useful as clinical applicability of angiogenic growth fa ctor gene transfer, (C) 2001 Academic Press.