Amino Acids

, Volume 42, Issue 2, pp 887–898

Polyamine analogs modulate gene expression by inhibiting lysine-specific demethylase 1 (LSD1) and altering chromatin structure in human breast cancer cells

Authors

  • Qingsong Zhu
    • The Sidney Kimmel Comprehensive Cancer Center at Johns HopkinsThe Johns Hopkins University School of Medicine
  • Yi Huang
    • Department of Pharmacology and Chemical BiologyUniversity of Pittsburgh Cancer Institute
  • Laurence J. Marton
    • Department of Laboratory MedicineThe University of California, San Francisco School of Medicine
  • Patrick M. Woster
    • Department of Pharmaceutical and Biomedical SciencesMedical University of South Carolina
  • Nancy E. Davidson
    • Department of Pharmacology and Chemical BiologyUniversity of Pittsburgh Cancer Institute
    • The Sidney Kimmel Comprehensive Cancer Center at Johns HopkinsThe Johns Hopkins University School of Medicine
    • Department of OncologyJohns Hopkins University
Original Article

DOI: 10.1007/s00726-011-1004-1

Cite this article as:
Zhu, Q., Huang, Y., Marton, L.J. et al. Amino Acids (2012) 42: 887. doi:10.1007/s00726-011-1004-1

Abstract

Aberrant epigenetic repression of gene expression has been implicated in most cancers, including breast cancer. The nuclear amine oxidase, lysine-specific demethylase 1 (LSD1) has the ability to broadly repress gene expression by removing the activating mono- and di-methylation marks at the lysine 4 residue of histone 3 (H3K4me1 and me2). Additionally, LSD1 is highly expressed in estrogen receptor α negative (ER−) breast cancer cells. Since epigenetic marks are reversible, they make attractive therapeutic targets. Here we examine the effects of polyamine analog inhibitors of LSD1 on gene expression, with the goal of targeting LSD1 as a therapeutic modality in the treatment of breast cancer. Exposure of the ER-negative human breast cancer cells, MDA-MB-231 to the LSD1 inhibitors, 2d or PG11144, significantly increases global H3K4me1 and H3K4me2, and alters gene expression. Array analysis indicated that 98 (75 up and 23 down) and 477 (237 up and 240 down) genes changed expression by at least 1.5-fold or greater after treatment with 2d and PG11144, respectively. The expression of 12 up-regulated genes by 2d and 14 up-regulated genes by PG11144 was validated by quantitative RT-PCR. Quantitative chromatin immunoprecipitation (ChIP) analysis demonstrated that up-regulated gene expression by polyamine analogs is associated with increase of the active histone marks H3K4me1, H3K4me2 and H3K9act, and decrease of the repressive histone marks H3K9me2 and H3K27me3, in the promoter regions of the relevant target genes. These data indicate that the pharmacologic inhibition of LSD1 can effectively alter gene expression and that this therapeutic strategy has potential.

Keywords

EpigeneticsChromatinHistone methylationAcetylationGene silencing

Abbreviations

Aza

5-Azacytidine

LSD1

Lysine-specific demethylase 1

DNMT

DNA methyltransferase

GAPDH

Glyceraldehyde 3-phosphate dehydrogenase

HDAC

Histone deacetylase

ODC

Ornithine decarboxylasae

H3K4

Lysine 4 of histone 3

H3K9act

Acetylated lysine 9 of histone 3

H3K27

Lysine 27 of histone 3

me1

Monomethyl

me2

Dimethyl

me3

Trimethyl

ChIP

Chromatin immunoprecipitation

mt1f

Metallothionein 1F

nupr

Nuclear protein/transcription regulator 1

egr1

Early growth response 1

cdh16

Cadherin 16/KSP-cadherin

Supplementary material

726_2011_1004_MOESM1_ESM.pdf (348 kb)
Fig. S1. Representative gene networks altered by 2d or PG11144. The networks altered by 2d corresponds functionally to cell death, gene expression and carbohydrate metabolism (A); cell cycle, cellular growth and proliferation and cell death (B); cellular growth and proliferation, hematological system development and function, inflammatory response (C). PG11144 changes lipid metabolism, small molecule biochemistry, vitamin and mineral metabolism (D); cell cycle, nutritional disease, cardiovascular disease (E); cancer, cell cycle and cellular movement networks (F). Supplementary material 1 (PDF 349 kb)

Copyright information

© Springer-Verlag 2011