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

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20 Citations (Scopus)


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
Issue number8
Publication statusPublished - 1999
Externally publishedYes


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

ASJC Scopus subject areas

  • Immunology and Allergy
  • Rheumatology
  • Immunology


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