A Novel Recombinant Basic Fibroblast Growth Factor and Its Secretion

• Published in: Biochemical and biophysical research communications 2001-06, Vol.284 (4), p.931-936
• Main Authors: Sohn, Young-Doug, Lim, Hyun Joung, Hwang, Ki-Chul, Kwon, Jun Hye, Park, Hyun-Young, Chung, Kwang-Hoe, Cho, Seung Yun, Jang, Yangsoo
• Format: Article
• Language: English
• Subjects: 3T3 Cells; Animals; Aorta, Thoracic - cytology; Aorta, Thoracic - drug effects; Aorta, Thoracic - physiology; basic fibroblast growth factor; Cell Division - drug effects; Cells, Cultured; Cercopithecus aethiops; Cloning, Molecular - methods; COS Cells; DNA - biosynthesis; Dose-Response Relationship, Drug; Fibroblast Growth Factor 2 - biosynthesis; Fibroblast Growth Factor 2 - genetics; Fibroblast Growth Factor 2 - pharmacology; Fibroblast Growth Factor 4; Fibroblast Growth Factors - genetics; Fibroblast Growth Factors - pharmacology; Fibroblasts - metabolism; Humans; Mice; Muscle, Smooth, Vascular - cytology; Muscle, Smooth, Vascular - drug effects; Muscle, Smooth, Vascular - physiology; Polymerase Chain Reaction; Proto-Oncogene Proteins - genetics; Proto-Oncogene Proteins - pharmacology; Rats; Rats, Sprague-Dawley; recombinant; Recombinant Fusion Proteins - pharmacology; Recombinant Proteins - biosynthesis; Recombinant Proteins - pharmacology; Reverse Transcriptase Polymerase Chain Reaction; secretion; Skin - metabolism; Transfection - methods
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1006/bbrc.2001.5076

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 applications for FGF-2 gene transfer are being evaluated in wound healing and collateral artery development to relieve myocardial and peripheral ischemia due to the ability of FGF-2 to regulate the growth and function of vascular cells. However, FGF-2 lacks a classical hydrophobic secretion signal peptide, the FGF-2 chimeras containing various signal sequences have been explored. In 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 increase the secretion level of FGF-2. When the recombinant FGF-2 gene, cloned into the expression vector with CMV promoter, was expressed in COS-7 cells, the recombinant 4sFGF-2 was highly secreted into the media. The secreted 4sFGF-2 showed the same biological activity as the native FGF-2 in the dose-response effects on DNA synthesis and cell growth of rat aortic smooth muscle 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 recombinant 4sFGF-2 may be useful as clinical applicability of angiogenic growth factor gene transfer.