370 DEVELOPMENT OF HYPOXIA-RESPONSIVE INTERFERON ALPHA 2B GENE CONSTRUCTS AND THEIR SUPPRESSIVE EFFECTS ON HUMAN RENAL CELL CARCINOMA CELL LINES IN VITRO

Abstract

You have accessJournal of UrologyKidney Cancer: Basic Research III1 Apr 2010370 DEVELOPMENT OF HYPOXIA-RESPONSIVE INTERFERON ALPHA 2B GENE CONSTRUCTS AND THEIR SUPPRESSIVE EFFECTS ON HUMAN RENAL CELL CARCINOMA CELL LINES IN VITRO Naotaka Fukui, Yukio Kageyama, Yusuke Kohno, Masaharu Inoue, Yotsuo Higashi, Shinae Kizaka-Kondoh, and Masahiro Hiraoka Naotaka FukuiNaotaka Fukui Ina town, Kita-adachigun, Saitama, Japan More articles by this author , Yukio KageyamaYukio Kageyama Ina town, Kita-adachigun, Saitama, Japan More articles by this author , Yusuke KohnoYusuke Kohno Ina town, Kita-adachigun, Saitama, Japan More articles by this author , Masaharu InoueMasaharu Inoue Ina town, Kita-adachigun, Japan More articles by this author , Yotsuo HigashiYotsuo Higashi Ina town, Kita-adachigun, Saitama, Japan More articles by this author , Shinae Kizaka-KondohShinae Kizaka-Kondoh Nishikyo-ku, Kyoto, Japan More articles by this author , and Masahiro HiraokaMasahiro Hiraoka Nishikyo-ku, Kyoto, Japan More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2010.02.437AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Interferon-α (IFNα) is one of the standard therapies for advanced renal cell carcinoma(RCC), though the response rate is low at 15-20 %. To improve effects of IFNα-based treatment for RCC, we invented a novel strategy using hypoxia-responsive IFN α2b gene constructs. METHODS We constructed three types of expression plasmid containing the coding region of the IFN α2b gene, pEF/5HRE.CMVmp-nls-ODD-IFNa2b (plasmid 1), pGL3/5HRE.CMVmp-IFNa 2b (plasmid 2), and pGL3/nls-ODD-IFNa2b (plasmid 3). Repetitive hypoxia response element (HRE) sequences and/or an oxygen-dependent degradation domain (ODD) has been inserted upstream of the IFN α2b gene. These IFNα2b gene constructs were transfected into human RCC cell lines, ACHN, SW839, 786-O, 769-P, caki-1, caki-2, KU19-20, KU20-01, and KU20-02. Concentration of IFN α2b in conditioned medium was measured by enzyme-linked immunosorbent assay. Viability of cells was determined by MTS assay. RESULTS All hypoxia-responsive IFN α2b gene constructs showed significant anti-proliferative effect on all RCC cell lines examined in this study. Deferoxamine mesilate (DFO), which mimics hypoxia, significantly enhanced the suppressive effects on ACHN, SW839, 786-O, KU19-20, and KU20-02 cells (Fig. 1). Secretion of IFNα was confirmed in conditioned media of KU20-01 media cells transfected with each construct. Plasmid 2 was most active in production of IFN, which was enhanced by DFO (Fig. 2). CONCLUSIONS Gene therapy using hypoxia-responsive interferon alpha 2b gene constructs may be a promising new treatment modality for renal cell carcinoma. © 2010 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 183Issue 4SApril 2010Page: e146-e147 Advertisement Copyright & Permissions© 2010 by American Urological Association Education and Research, Inc.MetricsAuthor Information Naotaka Fukui Ina town, Kita-adachigun, Saitama, Japan More articles by this author Yukio Kageyama Ina town, Kita-adachigun, Saitama, Japan More articles by this author Yusuke Kohno Ina town, Kita-adachigun, Saitama, Japan More articles by this author Masaharu Inoue Ina town, Kita-adachigun, Japan More articles by this author Yotsuo Higashi Ina town, Kita-adachigun, Saitama, Japan More articles by this author Shinae Kizaka-Kondoh Nishikyo-ku, Kyoto, Japan More articles by this author Masahiro Hiraoka Nishikyo-ku, Kyoto, Japan More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...