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| Content Provider | Springer Nature Link |
|---|---|
| Author | Aumsuwan, Pranapda Khan, Shabana I. Khan, Ikhlas A. Avula, Bharathi Walker, Larry A. Helferich, William G. Katzenellenbogen, Benita S. Dasmahapatra, Asok K. |
| Copyright Year | 2014 |
| 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. |
| Starting Page | 59 |
| Ending Page | 71 |
| Page Count | 13 |
| File Format | |
| ISSN | 10712690 |
| Journal | In Vitro Cellular & Developmental Biology - Animal |
| Volume Number | 51 |
| Issue Number | 1 |
| e-ISSN | 1543706X |
| Language | English |
| Publisher | Springer US |
| Publisher Date | 2014-08-23 |
| Publisher Place | Boston |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Breast cancer Epigenetics DNMT TET Wild yam Methylation Cell Biology Developmental Biology Stem Cells Cell Culture Animal Genetics and Genomics |
| Content Type | Text |
| Resource Type | Article |
| Subject | Cell Biology Developmental Biology Medicine |
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