Methylation mediated silencing of TMS1 in breast cancer and its potential contribution to docetaxel cytotoxicity

Anticancer Res. 2009 Aug;29(8):3207-10.

Abstract

Background: The tumor suppressor gene TMS1 (target of methylation-induced silencing) has been described in the literature as a pro-apoptotic gene. This study examined the methylation status of TMS1 in breast cancer cells and its potential role in sensitivity to docetaxel chemotherapy.

Materials and methods: Methylation of the TMS1 promoter was examined by methylation-specific PCR (MS-PCR) and gene expression was analyzed by reverse transcriptase PCR (RT-PCR). Apoptosis was evaluated by annexin V/propidium iodide staining followed by flow cytometric analysis.

Results and conclusion: The TMS1 promoter was unmethylated in ZR75-1, MB-231 and MCF7 cells which expressed the gene and partially methylated in SKBR3 and Hs578t cells in which TMS1 expression was down-regulated. Treatment of SKBR3 and Hs578t cells with demethylating agents resulted in reactivation of the TMS1 gene. Pretreatment with 5-azacytidine increased sensitivity to docetaxel treatment in SKBR3 and Hs578t cells, indicating that TMS1 reactivation in these cells may contribute to docetaxel sensitivity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antineoplastic Agents / pharmacology*
  • Apoptosis / drug effects
  • Blotting, Western
  • Breast Neoplasms / drug therapy*
  • Breast Neoplasms / genetics*
  • CARD Signaling Adaptor Proteins
  • Cytoskeletal Proteins / genetics*
  • DNA Methylation*
  • Docetaxel
  • Female
  • Flow Cytometry
  • Gene Expression Regulation, Neoplastic / drug effects
  • Gene Silencing / drug effects*
  • Humans
  • Promoter Regions, Genetic / genetics
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Taxoids / pharmacology*
  • Tumor Cells, Cultured

Substances

  • Antineoplastic Agents
  • CARD Signaling Adaptor Proteins
  • Cytoskeletal Proteins
  • PYCARD protein, human
  • RNA, Messenger
  • Taxoids
  • Docetaxel