Cancer Chemotherapy and Pharmacology

, Volume 48, Issue 1, pp 71–76

Antineoplastic action of 5-aza-2′-deoxycytidine and histone deacetylase inhibitor and their effect on the expression of retinoic acid receptor β and estrogen receptor α genes in breast carcinoma cells

  • Veronica Bovenzi
  • Richard L. Momparler
Original Article

DOI: 10.1007/s002800100294

Cite this article as:
Bovenzi, V. & Momparler, R.L. Cancer Chemother Pharmacol (2001) 48: 71. doi:10.1007/s002800100294


Purpose: During tumorigenesis several cancer-related genes can be silenced by aberrant methylation. In many cases these silenced genes can be reactivated by exposure to the DNA methylation inhibitor, 5-aza-2′-deoxycytidine (5-AZA-CdR). Histone acetylation also plays a role in the control of expression of some genes. The aim of this study was to determine the antineoplastic activities of 5-AZA-CdR and trichostatin A (TSA), either administered alone or in combination, in MDA-MB-231 breast carcinoma cells. The effects of these drugs (alone and in combination) on the expression of the tumor suppressor gene, retinoic acid receptor (RARβ) and of the estrogen receptor α gene (ERα), whose expression is lost in the cell line used in the study, were also investigated. Methods: MDA-MB-231 cells were treated with 5-AZA-CdR and TSA and the antitumor activity of these drugs was determined by clonogenic assay. Total RNA was extracted from the treated cells and RT-PCR was used to determine the effect of the treatment on the expression of RARβ and ERα. Methylation-sensitive PCR analysis was used to confirm that lack of expression of both genes was due to hypermethylation of their promoter regions. A single nucleotide primer extension assay was also used to quantify the reduction in DNA methylation following drug treatment. Results: Both 5-AZA-CdR and TSA alone showed significant antineoplastic activity. The combination of the two drugs was synergistic with respect to MDA-MB-231 cell kill. 5-AZA-CdR alone weakly activated the expression of both RARβ and ERα. TSA alone only activated RARβ, but not ERα. The combination of these agents appeared to produce a greater activation of both genes. Conclusions: The interesting interaction between 5-AZA-CdR and TSA in both cell kill and cancer-related gene reactivation provides a rationale for the use of inhibitors of DNA methylation and histone deacetylation in combination for the chemotherapy of breast cancer.

Breast cancer 5-Aza-2′-deoxycytidine Trichostatin A Retinoic acid receptor β Estrogen receptor α DNA methylation

Copyright information

© Springer-Verlag 2001

Authors and Affiliations

  • Veronica Bovenzi
    • 1
  • Richard L. Momparler
    • 2
  1. 1.Département de pharmacologie, Université de Montréal, Québec, Canada
  2. 2.Centre de recherche pédiatrique, Hôpital Ste-Justine, 3175 Côte Ste-Catherine, Montréal, Québec H3T 1C5, Canada