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 language | English (US) |
|---|---|
| Pages (from-to) | 59-71 |
| Number of pages | 13 |
| Journal | In Vitro Cellular and Developmental Biology - Animal |
| Volume | 51 |
| Issue number | 1 |
| DOIs | |
| State | Published - Jan 1 2014 |
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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 journal › Article
}
TY - JOUR
T1 - Evaluation of wild yam (Dioscorea villosa) root extract as a potential epigenetic agent in breast cancer cells
AU - Aumsuwan, Pranapda
AU - Khan, Shabana I.
AU - Khan, Ikhlas A.
AU - Avula, Bharathi
AU - Walker, Larry A.
AU - Helferich, William G
AU - Katzenellenbogen, Benita S
AU - Dasmahapatra, Asok K.
PY - 2014/1/1
Y1 - 2014/1/1
N2 - 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.
AB - 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.
KW - Breast cancer
KW - DNMT
KW - Epigenetics
KW - Methylation
KW - TET
KW - Wild yam
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UR - http://www.scopus.com/inward/citedby.url?scp=84961290228&partnerID=8YFLogxK
U2 - 10.1007/s11626-014-9807-5
DO - 10.1007/s11626-014-9807-5
M3 - Article
VL - 51
SP - 59
EP - 71
JO - In Vitro Cellular and Developmental Biology - Animal
JF - In Vitro Cellular and Developmental Biology - Animal
SN - 1071-2690
IS - 1
ER -