Control of delivered gene expression in chondrocytes using heat shock protein 70B promoter

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

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Objective. To investigate whether the expressions of delivered Escherichia coli β-galactosidase (LacZ) gene and transforming growth factor- β1 (TGF-β1) gene are regulated by the stress response of human chondrocyte- like cells (HCS-2/8) when heat shock protein 70B (HSP70B) promoter is inserted into the adenovirus vector. Methods. Two adenovirus vectors that contain either LacZ gene or TGF-β1 gene regulated by HSP70B promoter were constructed. One of the adenovirus vectors was added to the culture of HCS2/8 and gene transduced cells were produced. We applied heat stress (43°C) to the transduced cells for 2 h and examined whether the expression of transduced LacZ and TGF-β1 genes is affected by the stress, using 5 bromo-4- chloro-3-indolyl-β-D-galactopyranoside (X-gal) staining, measurement of β- galactosidase activity, Northern blotting, and ELISA. Results. The percentage of X-gal positive stained cells in LacZ gene-delivered cells with heat stress was significantly higher than in controls (no heat stress). With heat stress, β-galactosidase activity increased significantly, and the band of exogenous TGF-β1 mRNA became more apparent and the expression was maintained during the 24 h monitoring period. TGF-β1 level in culture supernatant of TGF-β1 gene-delivered cells with heat stress (5477.3 ± 321.1 pg/ml) was significantly higher than in the controls (853.2 ±29.2 pg/ml). Conclusion. HSP70B promoter could regulate the expression of delivered genes according to the intensity of heat stress.

Original languageEnglish
Pages (from-to)1769-1774
Number of pages6
JournalJournal of Rheumatology
Volume26
Issue number8
Publication statusPublished - 1999
Externally publishedYes

Keywords

  • Adenovirus vector
  • Gene
  • Heat shock protein
  • Osteoarthritis
  • Promoter
  • Transforming growth factor-β1

ASJC Scopus subject areas

  • Rheumatology
  • Immunology and Allergy
  • Immunology

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    Arai, Y., Kubo, T., Kobayashi, K., Ikeda, T., Takahashi, K., Takigawa, M., Imanishi, J., & Hirasawa, Y. (1999). Control of delivered gene expression in chondrocytes using heat shock protein 70B promoter. Journal of Rheumatology, 26(8), 1769-1774.