Anti-angiogenic effect of an insertional fusion protein of human basic fibroblast growth factor and ribonuclease-I

Tetsu Hayashida, Masakazu Ueda, Koichi Aiura, Hiroko Tada, Masayuki Onizuka, Masaharu Seno, Hidenori Yamada, Masaki Kitajima

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Human pancreatic ribonuclease-1 (RNase1) does not exhibit its cytotoxicity unless it is artificially internalized into the cytosol. Furthermore, once it encounters the cytosolic RNase inhibitor (RI), the activity of RNase1 is seriously reduced. To achieve the cellular targeting of RNase1 and the blocking of RI binding simultaneously, the basic fibroblast growth factor (bFGF) sequence was inserted into RNase1 at the RI binding site using a gene fusion technique. The effect of this fusion protein, CL-RFN89, on the angiogenesis, which was accelerated by FGF-FGF receptor interaction, was investigated. It was shown by using fluorescein-labeled CL-RFN89, that the binding to human umbilical vein endothelial cells (HUVECs) was dependent on the existence of the FGF receptors. In addition, CL-RFN89 inhibited the cellular growth of HUVECs in vitro and also inhibited the tube formation, using a three-dimensional tube formation assay. Furthermore, this fusion protein was shown to prevent in vivo tumor cell-induced angiogenesis, using the mouse dorsal air sac assay. These results demonstrated that CL-RFN89 inhibits angiogenesis in vitro and in vivo and that it can be expected to be a potent antiangiogenic agent.

Original languageEnglish
Pages (from-to)321-327
Number of pages7
JournalProtein Engineering, Design and Selection
Volume18
Issue number7
DOIs
Publication statusPublished - Jul 2005

Fingerprint

Pancreatic Ribonuclease
Fibroblast Growth Factor 2
Fibroblasts
Ribonucleases
Fusion reactions
Proteins
Fibroblast Growth Factor Receptors
Endothelial cells
Human Umbilical Vein Endothelial Cells
Assays
Air Sacs
Angiogenesis Inhibitors
Gene Fusion
Cytotoxicity
Fluorescein
Cytosol
Binding sites
Intercellular Signaling Peptides and Proteins
Tumors
Genes

Keywords

  • Angiogenesis
  • Basic fibroblast growth factor
  • Fusion protein
  • Molecular targeting
  • Ribonuclease

ASJC Scopus subject areas

  • Biochemistry
  • Biotechnology

Cite this

Anti-angiogenic effect of an insertional fusion protein of human basic fibroblast growth factor and ribonuclease-I. / Hayashida, Tetsu; Ueda, Masakazu; Aiura, Koichi; Tada, Hiroko; Onizuka, Masayuki; Seno, Masaharu; Yamada, Hidenori; Kitajima, Masaki.

In: Protein Engineering, Design and Selection, Vol. 18, No. 7, 07.2005, p. 321-327.

Research output: Contribution to journalArticle

Hayashida, Tetsu ; Ueda, Masakazu ; Aiura, Koichi ; Tada, Hiroko ; Onizuka, Masayuki ; Seno, Masaharu ; Yamada, Hidenori ; Kitajima, Masaki. / Anti-angiogenic effect of an insertional fusion protein of human basic fibroblast growth factor and ribonuclease-I. In: Protein Engineering, Design and Selection. 2005 ; Vol. 18, No. 7. pp. 321-327.
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