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http://dx.doi.org/10.14348/molcells.2014.0154

TAp73 and ΔNp73 Have Opposing Roles in 5-aza-2'-Deoxycytidine-Induced Apoptosis in Breast Cancer Cells  

Lai, Jing (Department of Medical Oncology, Jinling Hospital, School of Medicine, Southern Medical University)
Yang, Fang (Department of Medical Oncology, Jinling Hospital, Medical School of Nanjing University)
Zhang, Wenwen (Department of Medical Oncology, Jinling Hospital, Medical School of Nanjing University)
Wang, Yanru (Department of Medical Oncology, Jinling Hospital, School of Medicine, Southern Medical University)
Xu, Jing (Department of Medical Oncology, Jinling Hospital, Medical School of Nanjing University)
Song, Wei (Department of Medical Oncology, Jinling Hospital, School of Medicine, Southern Medical University)
Huang, Guichun (Department of Medical Oncology, Jinling Hospital, Medical School of Nanjing University)
Gu, Jun (Department of General Surgery, Jinling Hospital, Medical School of Nanjing University)
Guan, Xiaoxiang (Department of Medical Oncology, Jinling Hospital, School of Medicine, Southern Medical University)
Publication Information
Molecules and Cells / v.37, no.8, 2014 , pp. 605-612 More about this Journal
Abstract
The p73 gene contains an extrinsic P1 promoter and an intrinsic P2 promoter, controlling the transcription of the pro-apoptotic TAp73 isoform and the anti-apoptotic ${\Delta}Np73$ isoform, respectively. The DNA methylation status of both promoters act equally in the epigenetic transcriptional regulation of their relevant isoforms. The aim of this study was to analyze the different effects of these p73 isoforms in 5-aza-2'-deoxycytidine (5-aza-dC)-induced apoptosis in breast cancer cells. We investigated the effects of the DNA demethylation agent, 5-aza-dC, on the T-47D breast cancer cell line, and evaluated the methylation status of the p73 promoters and expression of TAp73 and ${\Delta}Np73$. Furthermore, we assessed the expression of p53 and p73 isoforms in 5-aza-dC-treated T-47D cells and p53 knockout cells. 5-aza-dC induced significant anti-tumor effects in T-47D cells, including inhibition of cell viability, G1 phase arrest and apoptosis. This was associated with p73 promoter demethylation and a concomitant increase in TAp73 mRNA and protein expression. In contrast, the methylation status of promoter P2 was not associated with ${\Delta}Np73$ mRNA or protein levels. Furthermore, demethylation of P2 failed to inhibit the expression of ${\Delta}Np73$ with 5-aza-dC in the p53 knockdown cell model. Our study suggests that demethylation of the P1 and P2 promoters has opposite effects on the expression of p73 isoforms, namely up-regulation of TAp73 and down-regulation of ${\Delta}Np73$. We also demonstrate that p53 likely contributes to 5-aza-dC-induced ${\Delta}Np73$ transcriptional inactivation in breast cancer cells.
Keywords
5-aza-2'-deoxycytidine; breast cancer; methylation; p53; p73;
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