Mechanisms of the growth-inhibitory effect of the RNase-EGF fused protein against EGFR-overexpressing cells

Sojun Hoshimoto, Masakazu Ueda, Hiromitsu Jinno, Masaki Kitajima, Junichiro Futami, Masaharu Seno

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Background: We previously showed the usefulness of a fused protein of human pancreatic ribonuclease1 (hRNase1) with human epidermal growth factor (hEGF) for molecular targeting of EGF receptor (EGFR)-overexpressing cells. In this study, the mechanisms underlying the inhibition of cell growth by RNase-EGF fused proteins was confirmed. Materials and Methods: Des.1-7 hRNase1 was genetically fused to hEGF. The fused proteins were expressed and isolated from Escherichia coli. The internalization of hRNase1-hEGF was confirmed by confocal fluorescence microscopy. The growth-inhibitory effect of the fused proteins was evaluated by MTT assay. Results: FITC-labelled hRNase1-hEGF was internalized into EGFR-overexpressing A431 cells. The internalization was not observed in A431 cells pre-treated with hEGF and EGFR-deficient H69 cells. The growth-inhibitory effect of des.1-7 hRNase1-hEGF against A431 cells was statistically significantly more pronounced than that of hRNase1-hEGF. Conclusion: RNase-EGF fused proteins are internalized through EGFR and inhibit cell growth by exerting their ribonucleolytic activity in the cytosol.

Original languageEnglish
Pages (from-to)857-863
Number of pages7
JournalAnticancer Research
Volume26
Issue number2 A
Publication statusPublished - Mar 2006

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Ribonucleases
Epidermal Growth Factor Receptor
Epidermal Growth Factor
Growth
Proteins
Fluorescein-5-isothiocyanate
Fluorescence Microscopy
Confocal Microscopy
Cytosol
Escherichia coli

Keywords

  • EGF receptor
  • Internalization
  • Ribonuclease
  • Targeting

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Mechanisms of the growth-inhibitory effect of the RNase-EGF fused protein against EGFR-overexpressing cells. / Hoshimoto, Sojun; Ueda, Masakazu; Jinno, Hiromitsu; Kitajima, Masaki; Futami, Junichiro; Seno, Masaharu.

In: Anticancer Research, Vol. 26, No. 2 A, 03.2006, p. 857-863.

Research output: Contribution to journalArticle

Hoshimoto, Sojun ; Ueda, Masakazu ; Jinno, Hiromitsu ; Kitajima, Masaki ; Futami, Junichiro ; Seno, Masaharu. / Mechanisms of the growth-inhibitory effect of the RNase-EGF fused protein against EGFR-overexpressing cells. In: Anticancer Research. 2006 ; Vol. 26, No. 2 A. pp. 857-863.
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AU - Futami, Junichiro

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