Breast Cancer Research and Treatment

, Volume 131, Issue 3, pp 777–789

Inhibitors of histone demethylation and histone deacetylation cooperate in regulating gene expression and inhibiting growth in human breast cancer cells

  • Yi Huang
  • Shauna N. Vasilatos
  • Lamia Boric
  • Patrick G. Shaw
  • Nancy E. Davidson
Preclinical Study

Abstract

Abnormal activities of histone lysine demethylases (KDMs) and lysine deacetylases (HDACs) are associated with aberrant gene expression in breast cancer development. However, the precise molecular mechanisms underlying the crosstalk between KDMs and HDACs in chromatin remodeling and regulation of gene transcription are still elusive. In this study, we showed that treatment of human breast cancer cells with inhibitors targeting the zinc cofactor dependent class I/II HDAC, but not NAD+ dependent class III HDAC, led to significant increase of H3K4me2 which is a specific substrate of histone lysine-specific demethylase 1 (LSD1) and a key chromatin mark promoting transcriptional activation. We also demonstrated that inhibition of LSD1 activity by a pharmacological inhibitor, pargyline, or siRNA resulted in increased acetylation of H3K9 (AcH3K9). However, siRNA knockdown of LSD2, a homolog of LSD1, failed to alter the level of AcH3K9, suggesting that LSD2 activity may not be functionally connected with HDAC activity. Combined treatment with LSD1 and HDAC inhibitors resulted in enhanced levels of H3K4me2 and AcH3K9, and exhibited synergistic growth inhibition of breast cancer cells. Finally, microarray screening identified a unique subset of genes whose expression was significantly changed by combination treatment with inhibitors of LSD1 and HDAC. Our study suggests that LSD1 intimately interacts with histone deacetylases in human breast cancer cells. Inhibition of histone demethylation and deacetylation exhibits cooperation and synergy in regulating gene expression and growth inhibition, and may represent a promising and novel approach for epigenetic therapy of breast cancer.

Keywords

Histone demethylase Histone deacetylase Epigenetics Breast cancer Growth inhibition Gene expression 

Supplementary material

10549_2011_1480_MOESM1_ESM.docx (28 kb)
Supplementary material 1 (DOCX 28 kb)
10549_2011_1480_MOESM2_ESM.ppt (264 kb)
Supplementary material 2 (PPT 264 kb)

References

  1. 1.
    Stearns V, Zhou Q, Davidson NE (2007) Epigenetic regulation as a new target for breast cancer therapy. Cancer Invest 25(8):659–665PubMedCrossRefGoogle Scholar
  2. 2.
    Marks PA, Richon VM, Miller T, Kelly WK (2004) Histone deacetylase inhibitors. Adv Cancer Res 91:137–168PubMedCrossRefGoogle Scholar
  3. 3.
    Ficner R (2009) Novel structural insights into class I and II histone deacetylases. Curr Top Med Chem 9(3):235–240PubMedCrossRefGoogle Scholar
  4. 4.
    Keen JC, Yan L, Mack KM, Pettit C, Smith D, Sharma D, Davidson NE (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(3):177–186PubMedCrossRefGoogle Scholar
  5. 5.
    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(1):64–69PubMedCrossRefGoogle Scholar
  6. 6.
    Yang X, Ferguson AT, Nass SJ, Phillips DL, Butash KA, Wang SM, Herman JG, Davidson NE (2000) Transcriptional activation of estrogen receptor alpha in human breast cancer cells by histone deacetylase inhibition. Cancer Res 60(24):6890–6894PubMedGoogle Scholar
  7. 7.
    Sharma D, Saxena NK, Davidson NE, Vertino PM (2006) Restoration of tamoxifen sensitivity in estrogen receptor-negative breast cancer cells: tamoxifen-bound reactivated ER recruits distinctive corepressor complexes. Cancer Res 66(12):6370–6378PubMedCrossRefGoogle Scholar
  8. 8.
    Shi Y, Lan F, Matson C, Mulligan P, Whetstine JR, Cole PA, Casero RA, Shi Y (2004) Histone demethylation mediated by the nuclear amine oxidase homolog LSD1. Cell 119(7):941–953PubMedCrossRefGoogle Scholar
  9. 9.
    Lee MG, Wynder C, Cooch N, Shiekhattar R (2005) An essential role for CoREST in nucleosomal histone 3 lysine 4 demethylation. Nature 437(7057):432–435PubMedGoogle Scholar
  10. 10.
    Kahl P, Gullotti L, Heukamp LC, Wolf S, Friedrichs N, Vorreuther R, Solleder G, Bastian PJ, Ellinger J, Metzger E et al (2006) Androgen receptor coactivators lysine-specific histone demethylase 1 and four and a half LIM domain protein 2 predict risk of prostate cancer recurrence. Cancer Res 66(23):11341–11347PubMedCrossRefGoogle Scholar
  11. 11.
    Scoumanne A, Chen X (2007) The lysine-specific demethylase 1 is required for cell proliferation in both p53-dependent and -independent manners. J Biol Chem 282(21):15471–15475PubMedCrossRefGoogle Scholar
  12. 12.
    Bradley C, van der Meer R, Roodi N, Yan H, Chandrasekharan MB, Sun ZW, Mernaugh RL, Parl FF (2007) Carcinogen-induced histone alteration in normal human mammary epithelial cells. Carcinogenesis 28(10):2184–2192PubMedCrossRefGoogle Scholar
  13. 13.
    Huang Y, Greene E, Murray Stewart T, Goodwin AC, Baylin SB, Woster PM, Casero RA Jr (2007) Inhibition of lysine-specific demethylase 1 by polyamine analogues results in reexpression of aberrantly silenced genes. Proc Natl Acad Sci USA 104(19):8023–8028PubMedCrossRefGoogle Scholar
  14. 14.
    Huang Y, Stewart TM, Wu Y, Baylin SB, Marton LJ, Perkins B, Jones RJ, Woster PM, Casero RA Jr (2009) Novel oligoamine analogues inhibit lysine-specific demethylase 1 and induce reexpression of epigenetically silenced genes. Clin Cancer Res 15(23):7217–7228PubMedCrossRefGoogle Scholar
  15. 15.
    Huang Y, Marton LJ, Woster PM, Casero RA (2009) Polyamine analogues targeting epigenetic gene regulation. Essays Biochem 46:95–110PubMedCrossRefGoogle Scholar
  16. 16.
    Karytinos A, Forneris F, Profumo A, Ciossani G, Battaglioli E, Binda C, Mattevi A (2009) A novel mammalian flavin-dependent histone demethylase. J Biol Chem 284(26):17775–17782PubMedCrossRefGoogle Scholar
  17. 17.
    Ciccone DN, Su H, Hevi S, Gay F, Lei H, Bajko J, Xu G, Li E, Chen T (2009) KDM1B is a histone H3K4 demethylase required to establish maternal genomic imprints. Nature 461(7262):415–418PubMedCrossRefGoogle Scholar
  18. 18.
    Yang Z, Jiang J, Stewart DM, Qi S, Yamane K, Li J, Zhang Y, Wong J (2010) AOF1 is a histone H3K4 demethylase possessing demethylase activity-independent repression function. Cell Res 20(3):276–287Google Scholar
  19. 19.
    Huang Y, Hager ER, Phillips DL, Dunn VR, Hacker A, Frydman B, Kink JA, Valasinas AL, Reddy VK, Marton LJ et al (2003) A novel polyamine analog inhibits growth and induces apoptosis in human breast cancer cells. Clin Cancer Res 9(7):2769–2777PubMedGoogle Scholar
  20. 20.
    Huang Y, Keen JC, Hager E, Smith R, Hacker A, Frydman B, Valasinas AL, Reddy VK, Marton LJ, Casero RA Jr et al (2004) Regulation of polyamine analogue cytotoxicity by c-Jun in human MDA-MB-435 cancer cells. Mol Cancer Res 2(2):81–88PubMedGoogle Scholar
  21. 21.
    Chou TC, Talalay P (1984) Quantitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme inhibitors. Adv Enzyme Regul 22:27–55PubMedCrossRefGoogle Scholar
  22. 22.
    Hahm HA, Dunn VR, Butash KA, Deveraux WL, Woster PM, Casero RA Jr, Davidson NE (2001) Combination of standard cytotoxic agents with polyamine analogues in the treatment of breast cancer cell lines. Clin Cancer Res 7(2):391–399PubMedGoogle Scholar
  23. 23.
    Tusher VG, Tibshirani R, Chu G (2001) Significance analysis of microarrays applied to the ionizing radiation response. Proc Natl Acad Sci USA 98(9):5116–5121PubMedCrossRefGoogle Scholar
  24. 24.
    Blander G, Guarente L (2004) The Sir2 family of protein deacetylases. Annu Rev Biochem 73:417–435PubMedCrossRefGoogle Scholar
  25. 25.
    Shi YJ, Matson C, Lan F, Iwase S, Baba T, Shi Y (2005) Regulation of LSD1 histone demethylase activity by its associated factors. Mol Cell 19(6):857–864PubMedCrossRefGoogle Scholar
  26. 26.
    Lan F, Collins RE, De Cegli R, Alpatov R, Horton JR, Shi X, Gozani O, Cheng X, Shi Y (2007) Recognition of unmethylated histone H3 lysine 4 links BHC80 to LSD1-mediated gene repression. Nature 448(7154):718–722PubMedCrossRefGoogle Scholar
  27. 27.
    Cameron EE, Bachman KE, Myohanen S, Herman JG, Baylin SB (1999) Synergy of demethylation and histone deacetylase inhibition in the re-expression of genes silenced in cancer. Nat Genet 21(1):103–107PubMedCrossRefGoogle Scholar
  28. 28.
    Gore SD, Baylin S, Sugar E, Carraway H, Miller CB, Carducci M, Grever M, Galm O, Dauses T, Karp JE et al (2006) Combined DNA methyltransferase and histone deacetylase inhibition in the treatment of myeloid neoplasms. Cancer Res 66(12):6361–6369PubMedCrossRefGoogle Scholar
  29. 29.
    Alexopoulou AN, Leao M, Caballero OL, Da Silva L, Reid L, Lakhani SR, Simpson AJ, Marshall JF, Neville AM, Jat PS (2010) Dissecting the transcriptional networks underlying breast cancer: NR4A1 reduces the migration of normal and breast cancer cell lines. Breast Cancer Res 12(4):R51Google Scholar
  30. 30.
    Wu Q, Dawson MI, Zheng Y, Hobbs PD, Agadir A, Jong L, Li Y, Liu R, Lin B, Zhang XK (1997) Inhibition of trans-retinoic acid-resistant human breast cancer cell growth by retinoid X receptor-selective retinoids. Mol Cell Biol 17(11):6598–6608PubMedGoogle Scholar
  31. 31.
    Novak P, Jensen T, Oshiro MM, Watts GS, Kim CJ, Futscher BW (2008) Agglomerative epigenetic aberrations are a common event in human breast cancer. Cancer Res 68(20):8616–8625PubMedCrossRefGoogle Scholar
  32. 32.
    Liang G, Bansal G, Xie Z, Druey KM (2009) RGS16 inhibits breast cancer cell growth by mitigating phosphatidylinositol 3-kinase signaling. J Biol Chem 284(32):21719–21727PubMedCrossRefGoogle Scholar
  33. 33.
    Rao R, Nalluri S, Kolhe R, Yang Y, Fiskus W, Chen J, Ha K, Buckley KM, Balusu R, Coothankandaswamy V et al (2010) Treatment with panobinostat induces glucose-regulated protein 78 acetylation and endoplasmic reticulum stress in breast cancer cells. Mol Cancer Ther 9(4):942–952Google Scholar
  34. 34.
    Ho TF, Ma CJ, Lu CH, Tsai YT, Wei YH, Chang JS, Lai JK, Cheuh PJ, Yeh CT, Tang PC et al (2007) Undecylprodigiosin selectively induces apoptosis in human breast carcinoma cells independent of p53. Toxicol Appl Pharmacol 225(3):318–328PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC. 2011

Authors and Affiliations

  • Yi Huang
    • 1
    • 2
  • Shauna N. Vasilatos
    • 1
  • Lamia Boric
    • 1
  • Patrick G. Shaw
    • 1
    • 3
  • Nancy E. Davidson
    • 1
    • 2
  1. 1.University of Pittsburgh Cancer InstitutePittsburghUSA
  2. 2.Department of Pharmacology & Chemical BiologyUniversity of PittsburghPittsburghUSA
  3. 3.Johns Hopkins University Bloomberg School of Public HealthBaltimoreUSA

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