Effects of the histone deacetylase inhibitor trichostatin A on nuclear texture and c-jun gene expression in drug-sensitive and drug-resistant human H69 lung carcinoma cells

Cytometry A. 2004 Dec;62(2):109-17. doi: 10.1002/cyto.a.20088.


Background: Texture analysis of chromatin patterns by image cytometry can be used in the development and refinement of diagnosis and prognosis of cancers and in the follow-up of therapies. However, little is known about the biological mechanisms underlying these patterns. Epigenetic mechanisms as histone posttranslational modifications and particularly histone acetylation could play a major role in the determination of these chromatin patterns and then influence nuclear texture measurements.

Methods: This study examined the consequences of treatment by the histone deacetylase inhibitor trichostatin A (TSA) on the nuclear texture in human cell lines sensitive and resistant to chemotherapy. Small cell lung carcinoma H69 cells and their variant H69-VP, which is resistant to etoposide, were incubated with 100 ng/ml of TSA for 0 to 24 h. Nuclear texture was evaluated by image cytometry and compared with the histone H4 acetylation level measured by western blotting and expression of c-jun gene evaluated by reverse transcription and real-time polymerase chain reaction.

Results: TSA treatment induced an increase in histone H4 acetylation level in both cell lines. However, at the level of chromatin texture, sensitive H69 cells displayed a progressive chromatin decondensation up to 24 h, whereas resistant H69-VP showed rapid (8 h) but transient changes. Similarly, expression of c-jun increased regularly in TSA-treated H69 cells. In H69-VP cells, an increase was also observed up to 12 h followed by a decrease after 24 h of treatment.

Conclusions: Analysis of nuclear texture appeared to be a sensitive technique to detect chromatin pattern alterations induced by the histone deacetylase inhibitor TSA in the H69 cell line and enabled the observation of chromatin pattern discrepancies between chemotherapeutic drug-sensitive and drug-resistant cells during this treatment. When c-jun gene expression was analyzed as gene sensitive to epigenetic control, these textural differences seemed to be correlated to gene expression.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carcinoma, Small Cell / genetics
  • Cell Line, Tumor
  • Cell Nucleus / drug effects*
  • Cell Survival / drug effects
  • Chromatin / drug effects
  • Drug Resistance, Neoplasm*
  • Enzyme Inhibitors / pharmacology*
  • Epigenesis, Genetic
  • Gene Expression / drug effects
  • Genes, jun / drug effects*
  • Histone Deacetylases / drug effects
  • Humans
  • Hydroxamic Acids / pharmacology*
  • Image Cytometry
  • Immunoblotting
  • Lung Neoplasms / genetics
  • Reverse Transcriptase Polymerase Chain Reaction


  • Chromatin
  • Enzyme Inhibitors
  • Hydroxamic Acids
  • trichostatin A
  • Histone Deacetylases