Evaluation of wild yam (Dioscorea villosa) root extract as a potential epigenetic agent in breast cancer cells

Pranapda Aumsuwan, Shabana I. Khan, Ikhlas A. Khan, Bharathi Avula, Larry A. Walker, William G. Helferich, Benita S. Katzenellenbogen, Asok K. Dasmahapatra

Research output: Contribution to journalArticle

Abstract

The present study was designed to evaluate the efficacy of wild yam root extract (WYRE) as a potential demethylating agent using two breast cancer cell lines, MCF-7 (estrogen receptor positive; ER+) and MDA-MB-231 (Estrogen receptor negative; ER), and a methylated gene, GATA3, as a potential marker of breast cancer development. The cells were treated with WYRE (0–50 μg/mL) for 72 h and used for viability, mRNA, and methylation analyses. WYRE significantly reduced viability of both cell lines and enhanced mRNA content of GATA3 in a concentration-dependent manner; however, DNMT mRNAs (DNMT1, 3A, 3B) were found to increase significantly only in MDA-MB-231 cells. Global DNA methylation, analyzed as 5′-methyl-2′-deoxycytidine (5-mC) and 5-hydroxymethylcytosine (5-hmC), showed a concentration-dependent enhancement of 5-mC with no alteration in 5-hmC level in MCF-7 cells; however, in MDA-MB-231 cells, in contrast to MCF-7 cells, 5-mC remained unaltered but 5-hmC reduced significantly in all WYRE concentrations (10–50 μg/mL) used in this study. Since 5-hmC is generated from 5-mC by ten-eleven-translocation (TET) enzymes, analysis of TET mRNAs (TET1, TET2, and TET3) in MDA-MB-231 cells indicated a concentration-dependent reduction in TET1 and induction of TET3; however, TET2 remained unaltered. No alterations in any of the TET mRNAs were found in MCF-7 cells. Methylation analysis of GATA3 promoter at specific locus indicates probable demethylating activity of WYRE in MDA-MB-231 cells. We conclude that activation of GATA3 gene in ER MDA-MB-231 cells may occur by altering DNA methylation pattern on the promoter region which may be different from the mechanisms operated in ER+ MCF-7 cells.

Original languageEnglish (US)
Pages (from-to)59-71
Number of pages13
JournalIn Vitro Cellular and Developmental Biology - Animal
Volume51
Issue number1
DOIs
StatePublished - Jan 1 2014

Keywords

  • Breast cancer
  • DNMT
  • Epigenetics
  • Methylation
  • TET
  • Wild yam

ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology

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