Adenovirus vector-mediated gene transduction to chondrocytes: In vitro evaluation of therapeutic efficacy of transforming growth factor-β 1 and heat shock protein 70 gene transduction

Yuji Arai, Toshikazu Kubo, Kappei Kobayashi, Kazushige Takeshita, Kenji Takahashi, Takumi Ikeda, Jiro Imanishi, Masaharu Takigawa, Yasusuke Hirasawa

Research output: Contribution to journalArticle

83 Citations (Scopus)

Abstract

Objective. To investigate the effects of adenovirus vector-mediated gene transduction of E. coli β-galactosidase (LacZ), transforming growth factor- β1 (TGF-β1), and heat shock protein 70 (HSP 70) on human chondrocyte-like cell line (HCS-2/8). Methods. We examined expression of transduced genes and their expression periods by 5 bromo-4-chloroindolyl-β-D-galactoside (X-gal) staining, Northern blotting. ELISA, and Western blotting. To assess the influence of TGF-β1 and HSP70 gene transduction, the expression of mRNA of type II collagen, proteoglycan core protein, matrix metalloproteinase 3 (MMP3) and tissue inhibitor of matrix metalloproteinase 1 were examined by Northern blotting. Results. Staining with X-gal indicated that the genes were transduced into a majority of the cells. Expression of the transduced genes in the cells was continued for at least 21 days. Transduction of TGF-β1 gene enhanced mRNA expression of type II collagen and proteoglycan core protein, but suppressed MMP3 mRNA expression in the cells. Expression of HSP70 was also high. Enhanced expression of HSP70 elevated mRNA expression of proteoglycan core protein. Conclusion. These results indicate adenovirus vector is useful in chondrocyte gene therapy, and it could be an efficient mediator of TGF-β1 and HSP70 gene transduction.

Original languageEnglish
Pages (from-to)1787-1795
Number of pages9
JournalJournal of Rheumatology
Volume24
Issue number9
Publication statusPublished - Sep 1997

Keywords

  • Adenovirus vector
  • Chondrocytes
  • Gene transduction
  • Heat shock protein 70
  • Transforming growth factor-β1

ASJC Scopus subject areas

  • Rheumatology
  • Immunology and Allergy
  • Immunology

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