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Mutant p53 binds to estrogen receptor negative promoter via DNMT1 and HDAC1 in MDA-MB-468 breast cancer cells

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Abstract

DNA methylation and histone deacetylation are two epigenetic mechanisms involved in the lack of estrogen receptor (ER) expression. Our previous studies demonstrated that mutant p53 along with repression complex proteins including DNMT1, HDAC1 and MeCP2 is associated with ER-negative promoter in MDA-MB-468 cells. To elucidate the molecular mechanism of estrogen receptor 1 (ESR1) gene silencing in these cells, we down-regulated DNMT1 and HDAC1 expression using siRNAs and studied the ability of DNMT1, HDAC1, MeCP2 and p53 in binding to ESR1 promoter CpG island. Our results showed that DNMT1 or HDAC1 down-regulation disassembled the repression complex proteins and mutant p53 from ER-negative promoter. The partial demethylation of ESR1 promoter and ER re-expression in down-regulated cells supports these findings. In vivo binding studies demonstrated that mutation of p53 protein in this cell line did not affect its binding capacity to DNMT1, HDAC1 and MeCP2 proteins. Our observations suggest that not only histone deacetylase activity of HDAC1 contributes to inactivation of methylated ESR1 gene but also HDAC1 presence on ESR1 promoter is important for assembly of DNMT1 in repression complex. In addition, our data revealed that mutant p53 protein binds to the promoter of ESR1 through direct interaction with HDAC1 and indirect interaction with DNMT1, MeCP2 proteins in the ER-negative MDA-MB-468 cells.

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References

  1. Sui M, Zhang H, Fan W (2011) The role of estrogen and estrogen receptors in chemoresistance. Curr Med Chem 18:4674–4683

    Article  PubMed  CAS  Google Scholar 

  2. Giacinti L, Claudio PP, Lopez M, Giordano A (2006) Epigenetic information and estrogen receptor alpha expression in breast cancer. Oncologist 11:1–8

    Article  PubMed  CAS  Google Scholar 

  3. Lo PK, Sukumar S (2008) Epigenomics and breast cancer. Pharmacogenomics 9:1879–1902

    Article  PubMed  CAS  Google Scholar 

  4. Yan L, Yang X, Davidson NE (2001) Role of DNA methylation and histone acetylation in steroid receptor expression in breast cancer. J Mammary Gland Biol Neoplasia 6:183–192

    Article  PubMed  CAS  Google Scholar 

  5. Sharma D, Blum J, Yang X et al (2005) Release of methyl CpG binding proteins and histone deacetylase 1 from the Estrogen receptor alpha (ER) promoter upon reactivation in ER-negative human breast cancer cells. Mol Endocrinol 19:1740–1751

    Article  PubMed  CAS  Google Scholar 

  6. Yan L, Nass SJ, Smith D et al (2003) Specific inhibition of DNMT1 by antisense oligonucleotides induces re-expression of estrogen receptor-alpha (ER) in ER-negative human breast cancer cell lines. Cancer Biol Ther 2:552–556

    PubMed  CAS  Google Scholar 

  7. Tajima S, Suetake I (1998) Regulation and function of DNA methylation in vertebrates. J Biochem 123:993–999

    Article  PubMed  CAS  Google Scholar 

  8. Chedin F, Lieber MR, Hsieh CL (2002) The DNA methyltransferase-like protein DNMT3L stimulates de novo methylation by Dnmt3a. Proc Natl Acad Sci USA 99:16916–16921

    Article  PubMed  CAS  Google Scholar 

  9. Ferguson AT, Lapidus RG, Baylin SB, Davidson NE (1995) Demethylation of the estrogen receptor gene in estrogen receptor-negative breast cancer cells can reactivate estrogen receptor gene expression. Cancer Res 55:2279–2283

    PubMed  CAS  Google Scholar 

  10. Dobosy JR, Selker EU (2001) Emerging connections between DNA methylation and histone acetylation. Cell Mol Life Sci 58:721–727

    Article  PubMed  CAS  Google Scholar 

  11. Yang X, Ferguson AT, Nass SJ et al (2000) Transcriptional activation of estrogen receptor alpha in human breast cancer cells by histone deacetylase inhibition. Cancer Res 60:6890–6894

    PubMed  CAS  Google Scholar 

  12. Keen JC, Yan L, Mack KM et al (2003) A novel histone deacetylase inhibitor, scriptaid, enhances expression of functional estrogen receptor alpha (ER) in ER negative human breast cancer cells in combination with 5-aza 2′-deoxycytidine. Breast Cancer Res Treat 81:177–186

    Article  PubMed  CAS  Google Scholar 

  13. Zhou Q, Atadja P, Davidson NE (2007) Histone deacetylase inhibitor LBH589 reactivates silenced estrogen receptor alpha (ER) gene expression without loss of DNA hypermethylation. Cancer Biol Ther 6:64–69

    Article  PubMed  CAS  Google Scholar 

  14. Robertson KD, Ait-Si-Ali S, Yokochi T et al (2000) DNMT1 forms a complex with Rb, E2F1 and HDAC1 and represses transcription from E2F-responsive promoters. Nat Genet 25:338–342

    Article  PubMed  CAS  Google Scholar 

  15. Rountree MR, Bachman KE, Baylin SB (2000) DNMT1 binds HDAC2 and a new co-repressor, DMAP1, to form a complex at replication foci. Nat Genet 25:269–277

    Article  PubMed  CAS  Google Scholar 

  16. Qin W, Leonhardt H, Pichler G (2011) Regulation of DNA methyltransferase 1 by interactions and modifications. Nucleus 2:392–402

    Article  PubMed  Google Scholar 

  17. Kimura H, Shiota K (2003) Methyl-CpG-binding protein, MeCP2, is a target molecule for maintenance DNA methyltransferase, Dnmt1. J Biol Chem 278:4806–4812

    Article  PubMed  CAS  Google Scholar 

  18. Angeloni SV, Martin MB, Garcia-Morales P et al (2004) Regulation of estrogen receptor-alpha expression by the tumor suppressor gene p53 in MCF-7 cells. J Endocrinol 180:497–504

    Article  PubMed  CAS  Google Scholar 

  19. Goh AM, Coffill CR, Lane DP (2011) The role of mutant p53 in human cancer. J Pathol 223:116–126

    Article  PubMed  CAS  Google Scholar 

  20. Akaogi K, Nakajima Y, Ito I et al (2009) KLF4 suppresses estrogen-dependent breast cancer growth by inhibiting the transcriptional activity of ERalpha. Oncogene 28:2894–2902

    Article  PubMed  CAS  Google Scholar 

  21. Shirley SH, Rundhaug JE, Tian J et al (2009) Transcriptional regulation of estrogen receptor-alpha by p53 in human breast cancer cells. Cancer Res 69:3405–3414

    Article  PubMed  CAS  Google Scholar 

  22. Rasti M, Arabsolghar R, Khatooni Z, Mostafavi-Pour Z (2012) p53 Binds to estrogen receptor 1 promoter in human breast cancer cells. Pathol Oncol Res 18:169–175

    Article  PubMed  CAS  Google Scholar 

  23. Suzuki M, Sunaga N, Shames DS et al (2004) RNA interference-mediated knockdown of DNA methyltransferase 1 leads to promoter demethylation and gene re-expression in human lung and breast cancer cells. Cancer Res 64:3137–3143

    Article  PubMed  CAS  Google Scholar 

  24. Keshelava N, Davicioni E, Wan Z et al (2007) Histone deacetylase 1 gene expression and sensitization of multidrug-resistant neuroblastoma cell lines to cytotoxic agents by depsipeptide. J Natl Cancer Inst 99:1107–1119

    Article  PubMed  CAS  Google Scholar 

  25. Rasti M, Grand RJ, Mymryk JS et al (2005) Recruitment of CBP/p300, TATA-binding protein, and S8 to distinct regions at the N terminus of adenovirus E1A. J Virol 79:5594–5605

    Article  PubMed  CAS  Google Scholar 

  26. Rasti M, Tavasoli P, Monabati A, Entezam M (2009) Association between HIC1 and RASSF1A promoter hypermethylation with MTHFD1 G1958A polymorphism and clinicopathological features of breast cancer in Iranian patients. Iran Biomed J 13:199–206

    PubMed  CAS  Google Scholar 

  27. Lapidus RG, Nass SJ, Butash KA et al (1998) Mapping of ER gene CpG island methylation-specific polymerase chain reaction. Cancer Res 58:2515–2519

    PubMed  CAS  Google Scholar 

  28. Hoque MO, Prencipe M, Poeta ML et al (2009) Changes in CpG islands promoter methylation patterns during ductal breast carcinoma progression. Cancer Epidemiol Biomarkers Prev 18:2694–2700

    Article  PubMed  CAS  Google Scholar 

  29. Zhao L, Wang L, Jin F et al (2009) Silencing of estrogen receptor alpha (ERalpha) gene by promoter hypermethylation is a frequent event in Chinese women with sporadic breast cancer. Breast Cancer Res Treat 117:253–259

    Article  PubMed  CAS  Google Scholar 

  30. Saxena NK, Sharma D (2010) Epigenetic reactivation of estrogen receptor: promising tools for restoring response to endocrine therapy. Mol Cell Pharmacol 2:191–202

    PubMed  CAS  Google Scholar 

  31. Vinyals A, Peinado MA, Gonzalez-Garrigues M et al (1999) Failure of wild-type p53 gene therapy in human cancer cells expressing a mutant p53 protein. Gene Ther 6:22–33

    Article  PubMed  CAS  Google Scholar 

  32. Okano M, Bell DW, Haber DA, Li E (1999) DNA methyltransferases Dnmt3a and Dnmt3b are essential for de novo methylation and mammalian development. Cell 99:247–257

    Article  PubMed  CAS  Google Scholar 

  33. Yang X, Phillips DL, Ferguson AT et al (2001) Synergistic activation of functional estrogen receptor (ER)-alpha by DNA methyltransferase and histone deacetylase inhibition in human ER-alpha-negative breast cancer cells. Cancer Res 61:7025–7029

    PubMed  CAS  Google Scholar 

  34. Ng HH, Bird A (1999) DNA methylation and chromatin modification. Curr Opin Genet Dev 9:158–163

    Article  PubMed  CAS  Google Scholar 

  35. Harms K, Nozell S, Chen X (2004) The common and distinct target genes of the p53 family transcription factors. Cell Mol Life Sci 61:822–842

    Article  PubMed  CAS  Google Scholar 

  36. Ho JS, Ma W, Mao DY, Benchimol S (2005) p53-Dependent transcriptional repression of c-myc is required for G1 cell cycle arrest. Mol Cell Biol 25:7423–7431

    Article  PubMed  CAS  Google Scholar 

  37. Esteve PO, Chin HG, Pradhan S (2005) Human maintenance DNA (cytosine-5)-methyltransferase and p53 modulate expression of p53-repressed promoters. Proc Natl Acad Sci USA 102:1000–1005

    Article  PubMed  CAS  Google Scholar 

  38. Juan LJ, Shia WJ, Chen MH et al (2000) Histone deacetylases specifically down-regulate p53-dependent gene activation. J Biol Chem 275:20436–20443

    Article  PubMed  CAS  Google Scholar 

  39. Le Gac G, Esteve PO, Ferec C, Pradhan S (2006) DNA damage-induced down-regulation of human Cdc25C and Cdc2 is mediated by cooperation between p53 and maintenance DNA (cytosine-5) methyltransferase 1. J Biol Chem 281:24161–24170

    Article  PubMed  Google Scholar 

  40. Lai JC, Cheng YW, Goan YG et al (2008) Promoter methylation of O(6)-methylguanine-DNA-methyltransferase in lung cancer is regulated by p53. DNA Repair (Amst) 7:1352–1363

    Article  CAS  Google Scholar 

  41. Lim LY, Vidnovic N, Ellisen LW, Leong CO (2009) Mutant p53 mediates survival of breast cancer cells. Br J Cancer 101:1606–1612

    Article  PubMed  CAS  Google Scholar 

  42. Dhar G, Banerjee S, Dhar K et al (2008) Gain of oncogenic function of p53 mutants induces invasive phenotypes in human breast cancer cells by silencing CCN5/WISP-2. Cancer Res 68:4580–4587

    Article  PubMed  CAS  Google Scholar 

  43. Böhlig L, Rother K (2011) One function-multiple mechanisms: the manifold activities of p53 as a transcriptional repressor. J Biomed Biotechnol 2011:464916

    Article  PubMed  Google Scholar 

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Acknowledgments

This work was supported by Grants No: 89-5176 and 90-5620 from the Shiraz University of Medical Sciences, Iran for Ms Rita Arabsolghar’s thesis and Ms Tayebe Azimi’s thesis, respectively. The authors would like to thank the School of Advanced Medical Sciences and Technologies for their support in performing this project.

Conflict of interest

None of the authors has any conflict of interest.

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Authors and Affiliations

  1. Recombinant Protein Lab, Department of Biochemistry, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran

    Rita Arabsolghar, Tayebeh Azimi & Mozhgan Rasti

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  1. Rita Arabsolghar
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  2. Tayebeh Azimi
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  3. Mozhgan Rasti
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Corresponding author

Correspondence to Mozhgan Rasti.

Additional information

Rita Arabsolghar and Tayebeh Azimi contributed equally to this study and both are considered first author.

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Arabsolghar, R., Azimi, T. & Rasti, M. Mutant p53 binds to estrogen receptor negative promoter via DNMT1 and HDAC1 in MDA-MB-468 breast cancer cells. Mol Biol Rep 40, 2617–2625 (2013). https://doi.org/10.1007/s11033-012-2348-7

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  • DOI: https://doi.org/10.1007/s11033-012-2348-7

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