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Histone deacetylase inhibitor activates the p21/WAF1/Cip1 gene promoter through the Sp1 sites

Y Sowa1, T Orita, S Hiranabe-Minamikawa

  • 1Department of Preventive Medicine, Kyoto Prefectural University of Medicine, Japan.

Annals of the New York Academy of Sciences
|February 10, 2000
PubMed

Insights

Trichostatin A (TSA) triggers histone hyperacetylation, leading to cell growth arrest and increased p21/WAF1/Cip1 protein expression in MG63 cells. This activation occurs via Sp1 sites on the promoter, independent of p53.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Epigenetics

Background:

  • Trichostatin A (TSA) is a potent histone deacetylase inhibitor.
  • Histone acetylation plays a crucial role in gene expression regulation.
  • MG63 cells are a human osteosarcoma cell line commonly used in research.

Purpose of the Study:

  • To investigate the effects of Trichostatin A (TSA) on MG63 cell growth and gene expression.
  • To elucidate the mechanism by which TSA influences p21/WAF1/Cip1 protein induction.
  • To determine the role of specific promoter elements in TSA-mediated gene activation.

Main Methods:

  • Treatment of MG63 cells with TSA.
  • Analysis of cell growth and proliferation.
  • Western blotting to detect p21/WAF1/Cip1 protein levels.
  • Assessment of histone acetylation.
  • Reporter assays using mutated p21/WAF1/Cip1 promoter fragments.

Main Results:

  • TSA induced significant growth arrest in MG63 cells.
  • TSA treatment led to increased expression of the p21/WAF1/Cip1 protein.
  • A strong correlation was observed between histone hyperacetylation and p21/WAF1/Cip1 induction.
  • Mutation of Sp1 sites (-82 and -69) on the p21/WAF1/Cip1 promoter diminished the response to TSA.

Conclusions:

  • TSA activates the p21/WAF1/Cip1 promoter in MG63 cells.
  • The activation is mediated through specific Sp1 binding sites on the promoter.
  • The mechanism of TSA-induced p21/WAF1/Cip1 expression is independent of the p53 pathway.

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