| Autor: |
Huisman, Christian, van der Wijst, Monique GP, Falahi, Fahimeh, Overkamp, Juul, Karsten, Gellért, Terpstra, Martijn M, Kok, Klaas, van der Zee, Ate GJ, Schuuring, Ed, Wisman, G Bea A, Rots, Marianne G |
| Zdroj: |
Epigenetics; May 2015, Vol. 10 Issue: 5 p384-396, 13p |
| Abstrakt: |
Epigenetic silencing of tumor suppressor genes (TSGs) is considered a significant event in the progression of cancer. For example, EPB41L3, a potential biomarker in cervical cancer, is often silenced by cancer-specific promoter methylation. Artificial transcription factors (ATFs) are unique tools to re-express such silenced TSGs to functional levels; however, the induced effects are considered transient. Here, we aimed to improve the efficiency and sustainability of gene re-expression using engineered zinc fingers fused to VP64 (ZF-ATFs) or DNA methylation modifiers (ZF-Tet2 or ZF-TDG) and/or by co-treatment with epigenetic drugs [5-aza-2′-deoxycytidine or Trichostatin A (TSA)]. The EPB41L3-ZF effectively bound its methylated endogenous locus, as also confirmed by ChIP-seq. ZF-ATFs reactivated the epigenetically silenced target gene EPB41L3(∼10-fold) in breast, ovarian, and cervical cancer cell lines. Prolonged high levels of EPB41L3(∼150-fold) induction could be achieved by short-term co-treatment with epigenetic drugs. Interestingly, for otherwise ineffective ZF-Tet2 or ZF-TDG treatments, TSA facilitated re-expression of EPB41L3up to twofold. ATF-mediated re-expression demonstrated a tumor suppressive role for EPB41L3in cervical cancer cell lines. In conclusion, epigenetic reprogramming provides a novel way to improve sustainability of re-expression of epigenetically silenced promoters. |
| Databáze: |
Supplemental Index |
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