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

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5-methyldeoxycytidine
Dioscorea
Epigenomics
MCF-7 Cells
Breast Neoplasms
Messenger RNA
DNA Methylation
Estrogen Receptors
Methylation
Cell Line
Genetic Promoter Regions
Transcriptional Activation
5-hydroxymethylcytosine
Enzymes

Keywords

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

ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology

Cite this

Evaluation of wild yam (Dioscorea villosa) root extract as a potential epigenetic agent in breast cancer cells. / Aumsuwan, Pranapda; Khan, Shabana I.; Khan, Ikhlas A.; Avula, Bharathi; Walker, Larry A.; Helferich, William G; Katzenellenbogen, Benita S; Dasmahapatra, Asok K.

In: In Vitro Cellular and Developmental Biology - Animal, Vol. 51, No. 1, 01.01.2014, p. 59-71.

Research output: Contribution to journalArticle

Aumsuwan, Pranapda ; Khan, Shabana I. ; Khan, Ikhlas A. ; Avula, Bharathi ; Walker, Larry A. ; Helferich, William G ; Katzenellenbogen, Benita S ; Dasmahapatra, Asok K. / Evaluation of wild yam (Dioscorea villosa) root extract as a potential epigenetic agent in breast cancer cells. In: In Vitro Cellular and Developmental Biology - Animal. 2014 ; Vol. 51, No. 1. pp. 59-71.
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