Transformation of mouse BALB/c 3T3 cells with human basic fibroblast growth factor cDNA.

Sasada, Reiko; Kurokawa, Tsutomu; Iwane, Makoto; Igarashi, Koichi

doi:10.1128/mcb.8.2.588

Cited by 140 publications

(66 citation statements)

References 39 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Although the low efficiency may have resulted from masking of the epitopes of endogenous bFGF by the extracellular matrix [7], it did not provide direct evidence for the autocrine growth stimulation model. Consistent with our results, Sasada et al [17] reported that the morphologic transformation induced by the expression of human bFGF cDNA in mouse BALB/c3T3 was reversed by adding our anti-rbFGF antibodies. It was also concluded that bFGF is produced and stored in the transformed mouse fibroblast cells and that it exerts its mitogenic effect via receptors on the cell surface.…”

Section: Discussionsupporting

confidence: 92%

Antibodies against basic fibroblast growth factor inhibit the autocrine growth of pulmonary artery endothelial cells

et al. 1988

View full text Add to dashboard Cite

Anti-recombinant human basic fibroblast growth factor (rbFGF) antibodies effaeiently inhibited the basal proliferation of bovine pulmonary artery endothelial (BAE) cells. The cell-free extract of BAE cells stimulated the proliferation of bovine capillary endothelial cells and this activity was completely abolished by the antibodies. Furthermore, on heparin HPLC the activity was eluted at exactly the same retention time as that for authentic pituitary bFGF. These observations directly indicate that the BAE cells produce bFGF that stimulates their own basal growth by binding to specific receptors expressed on the cell surface.

show abstract

Section: Discussionsupporting

confidence: 92%

Antibodies against basic fibroblast growth factor inhibit the autocrine growth of pulmonary artery endothelial cells

et al. 1988

View full text Add to dashboard Cite

show abstract

“…Growth stimulation of mouse BALB/c3T3 cells was assayed as described previously [19]. Briefly, cells were plated on 96-well microtiter plates (2 x 103/well).…”

Section: Bioassaymentioning

confidence: 99%

Carboxyl‐terminal structure of basic fibroblast growth factor significantly contributes to its affinity for heparin

Seno

Sasada

Kurokawa

et al. 1990

European Journal of Biochemistry

View full text Add to dashboard Cite

The carboxyl-terminal sequence of basic fibroblast growth factor (bFGF) is rich in basic amino acid residues, a common characteristic amongst fibroblast growth factors, and is considered to contribute greatly to the binding to negatively charged extracellular matrixes such as heparin. To study the relationship between the affinity for heparin and the carboxyl-terminal structure of bFGF, amino-or carboxyl-terminal truncated molecules were produced in Escherichia coli using recombinant DNA techniques. These terminally truncated bFGFs were applied to a heparin-affinity HPLC column. Truncation of more than six amino acid residues from the carboxyl-terminal made the bFGF produced in E. coli markedly difficult to solubilize and weakened its affinity for heparin, though bFGF having up to 46 amino acids removed showed significant stimulation of the DNA synthesis of BALB/c3T3 cells. This stimulation of the DNA synthesis was also recognized by the bFGF having 40 amino acids removed from its amino-terminal, while the affinity of this peptide for heparin has been shown to be equal to that of the mature bFGF (146 amino acids). These results show that the affinity of bFGF for heparin depends significantly on its carboxyl-terminal structure and that the essential part for receptor binding is present between Asp4' and SerlOO. Moreover, it suggests that the Phe139Leu'40Pro141, present in all members of the FGF family, contributes greatly to the stable structure of the intact molecule. . Extraordinarily high affinity for heparin is one of the important characteristics of this family. In fact, this characteristic has been applied to the isolation and characterization of these proteins, especially basic FGF (bFGF) and acidic FGF (aFGF), from various tissues [15] and tumors [16]. The identification of specific fundamental domains of bFGF for receptor and heparin binding has been reported using synthetic polypeptides; these have suggested the existence of two domains for heparin binding and one for receptor binding [I 5 , 171. We isolated the cDNA for human bFGF and studied the expression of this cDNA in mammalian cells [18, 191 and in bacterial cells (201. We also purified and characterized the recombinant bFGF produced in Escherichia coli [20] and studied the contribution of the four cysteine residues to the protein conformation of bFGF through a site-directed mutagenesis procedure [21]. For further characterization, we constructed several bFGF molecules whose amino-or carboxyl-terminus was deleted. Here we report the structural characterization for heparin binding using these terminally truncated molecules.Correspondence to M . Seno, Biotechnology Research Laboratories, Research and Development Div., Takeda Chemical Industries Ltd, 2-17-85 Jusohonmachi, Yodogawa-ku, Osaka 532, JapanAbbreviations. F G F , fibroblast growth factor; bFGF, basic fibroblast growth factor; aFGF, acidic fibroblast growth factor.Enzymes. DNA polymerase I (EC 2.7.7.7); exonucleasc I11 (EC 3.1.11.2); lysozyme (EC 3.2.1.17); peroxidase (EC 1.11.1.7). MA...

show abstract

“…Much of the interest in these polypeptides stems from their ability to promote the growth of endothelial cells and to act as angiogenesis factors, and it has been suggested that such properties may play a role in the growth of solid tumors (12). Cultured fibroblasts treated with bFGF adopt characteristics that resemble those of transformed cells, although it remains contentious whether the cloned FGF gene or cDNA can function as a dominantly acting transforming gene (3,12,23,26,29).…”

mentioning

confidence: 99%

Detection and characterization of the fibroblast growth factor-related oncoprotein INT-2.

Dixon¹,

Deed²,

Acland³

et al. 1989

Mol. Cell. Biol.

View full text Add to dashboard Cite

Products of the fibroblast growth factor-related proto-oncogene int-2 have been detected by using a monoclonal antibody and polyclonal antisera raised against synthetic peptides predicted from the DNA sequence. COS-1 monkey cells transfected with int-2 DNA linked to the simian virus 40 early promoter contained at least four int-2-specific proteins, presumably representing modified forms of the expected 27-kilodalton primary translation product. The level of expression was increased approximately six- to eightfold by mutation of sequences around the presumed initiation codon, negating their capacity to encode a short oligopeptide in the +1 reading frame. Both tunicamycin inhibition and in vitro translation experiments indicated that some of the modifications correspond to asparagine-linked glycosylation, for which the sequence predicts a single site. In line with the similarities between INT-2 and other fibroblast growth factors, the in vitro translation products functioned as weak mitogens for mammary epithelial cells.

show abstract

Transformation of mouse BALB/c 3T3 cells with human basic fibroblast growth factor cDNA.

Cited by 140 publications

References 39 publications

Antibodies against basic fibroblast growth factor inhibit the autocrine growth of pulmonary artery endothelial cells

Antibodies against basic fibroblast growth factor inhibit the autocrine growth of pulmonary artery endothelial cells

Carboxyl‐terminal structure of basic fibroblast growth factor significantly contributes to its affinity for heparin

Detection and characterization of the fibroblast growth factor-related oncoprotein INT-2.

Contact Info

Product

Resources

About