Modulation of intracellular iron metabolism by iron chelation affects chromatin remodeling proteins and corresponding epigenetic modifications in breast cancer cells and increases their sensitivity to chemotherapeutic agents

Pogribny,Igor P.; Tryndyak,Volodymyr P.; Pogribna,Marta; Shpyleva,Svitlana; Surratt,Gordon; Gamboa da Costa,Gonçalo; Beland,Frederick A.

doi:10.3892/ijo.2013.1855

Modulation of intracellular iron metabolism by iron chelation affects chromatin remodeling proteins and corresponding epigenetic modifications in breast cancer cells and increases their sensitivity to chemotherapeutic agents

Authors:
- Igor P. Pogribny
- Volodymyr P. Tryndyak
- Marta Pogribna
- Svitlana Shpyleva
- Gordon Surratt
- Gonçalo Gamboa da Costa
- Frederick A. Beland
View Affiliations

Affiliations: Division of Biochemical Toxicology, National Center for Toxicological Research, FDA, Jefferson, AR 72079, USA
Published online on: March 12, 2013 https://doi.org/10.3892/ijo.2013.1855
Pages: 1822-1832

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Iron plays a vital role in the normal functioning of cells via the regulation of essential cellular metabolic reactions, including several DNA and histone-modifying proteins. The metabolic status of iron and the regulation of epigenetic mechanisms are well-balanced and tightly controlled in normal cells; however, in cancer cells these processes are profoundly disturbed. Cancer-related abnormalities in iron metabolism have been corrected through the use of iron-chelating agents, which cause an inhibition of DNA synthesis, G1-S phase arrest, an inhibition of epithelial-to-mesenchymal transition, and the activation of apoptosis. In the present study, we show that, in addition to these well-studied molecular mechanisms, the treatment of wild-type TP53 MCF-7 and mutant TP53 MDA-MB-231 human breast cancer cells with desferrioxamine (DFO), a model iron chelator, causes significant epigenetic alterations at the global and gene-specific levels. Specifically, DFO treatment decreased the protein levels of the histone H3 lysine 9 demethylase, Jumonji domain-containing protein 2A (JMJD2A), in the MCF-7 and MDA-MB-231 cells and down-regulated the levels of the histone H3 lysine 4 demethylase, lysine-specific demethylase 1 (LSD1), in the MDA-MB-231 cells. These changes were accompanied by alterations in corresponding metabolically sensitive histone marks. Additionally, we demonstrate that DFO treatment activates apoptotic programs in MCF-7 and MDA-MB-231 cancer cells and enhances their sensitivity to the chemotherapeutic agents, doxorubicin and cisplatin; however, the mechanisms underlying this activation differ. The induction of apoptosis in wild-type TP53 MCF-7 cells was p53-dependent, triggered mainly by the down-regulation of the JMJD2A histone demethylase, while in mutant TP53 MDA-MB-231 cells, the activation of the p53-independent apoptotic program was driven predominantly by the epigenetic up-regulation of p21.

View Figures

View References

1.	Teperino R, Schoonjans K and Auwerx J: Histone methyltransferases and demethylases; can they link metabolism and transcription? Cell Metab. 12:321–327. 2010. View Article : Google Scholar : PubMed/NCBI
2.	Burgio G, Onorati MC and Corona DFV: Chromatin remodeling regulation by small molecules and metabolites. Biochim Biophys Acta. 1799:671–680. 2010. View Article : Google Scholar : PubMed/NCBI
3.	Hou H and Yu H: Structural insights into histone lysine demethylation. Curr Opin Struct Biol. 20:739–748. 2010. View Article : Google Scholar : PubMed/NCBI
4.	Iyer LM, Abhiman S and Aravind L: Natural history of eukaryotic DNA methylation systems. Prog Mol Biol Transl Sci. 101:25–104. 2011. View Article : Google Scholar : PubMed/NCBI
5.	Tsukada Y-i, Fang J, Erdjument-Bromage H, Warren ME, Borchers CH, Tempst P and Zhang Y: Histone demethylation by a family of JmjC domain-containing proteins. Nature. 439:811–816. 2006. View Article : Google Scholar : PubMed/NCBI
6.	Yamane K, Toumazou C, Tsukada Y-i, Erdjument-Bromage H, Tempst P, Wong J and Zhang Y: JHDM2A, a JmjC-containing H3K9 demethylase, facilitates transcription activation by androgen receptor. Cell. 125:483–495. 2006. View Article : Google Scholar : PubMed/NCBI
7.	Wu H and Zhang Y: Mechanisms and functions of Tet protein–mediated 5-methylcytosine oxidation. Genes Dev. 25:2436–2452. 2011.
8.	McDonough MA, Loenarz C, Chowdhury R, Clifton IJ and Schofield CJ: Structural studies of human 2-oxoglutarate dependent oxygenases. Curr Opin Struct Biol. 20:659–672. 2010. View Article : Google Scholar
9.	Torti SV and Torti FM: Ironing out cancer. Cancer Res. 71:1511–1514. 2011. View Article : Google Scholar : PubMed/NCBI
10.	Jones PA and Baylin SB: The epigenomics of cancer. Cell. 128:683–692. 2007. View Article : Google Scholar : PubMed/NCBI
11.	Miller LD, Coffman LG, Chou JW, Black MA, Berg J, D’Agostino R Jr, Torti SV and Torti FM: An iron regulatory gene signature predicts outcome in breast cancer. Cancer Res. 71:6728–6737. 2011. View Article : Google Scholar : PubMed/NCBI
12.	Hanahan D and Weinberg RA: Hallmarks of cancer: the next generation. Cell. 144:646–674. 2011. View Article : Google Scholar : PubMed/NCBI
13.	Baylin SB and Jones PA: A decade of exploring the cancer epigenome - biological and translational implications. Nat Rev Cancer. 11:726–734. 2011. View Article : Google Scholar : PubMed/NCBI
14.	Buss JL, Torti FM and Torti SV: The role of iron chelation in cancer therapy. Curr Med Chem. 10:1021–1034. 2003. View Article : Google Scholar : PubMed/NCBI
15.	Yu Y, Gutierrez E, Kovacevic Z, Saletta F, Obeidy P, Suryo Rahmanto Y and Richardson DR: Iron chelators for the treatment of cancer. Curr Med Chem. 19:2689–2702. 2012. View Article : Google Scholar : PubMed/NCBI
16.	Kawamoto M, Horibe T, Kohno M and Kawakami K: A novel transferring receptor-targeted hybrid peptide disintegrates cancer cell membrane to induce rapid killing of cancer cells. BMC Cancer. 11:3592011. View Article : Google Scholar
17.	Rao VA, Klein SR, Agama KK, Toyoda E, Adachi N, Pommier Y and Shacter EB: The iron chelator Dp44mT causes DNA damage and selective inhibition of topoisomerase IIα in breast cancer cells. Cancer Res. 69:948–957. 2009.PubMed/NCBI
18.	Chekhun VF, Lukyanova NY, Kovalchuk O, Tryndyak VP and Pogribny IP: Epigenetic profiling of multidrug-resistant MCF-7 breast cancer adenocarcinoma cells reveals novel hyper- and hypomethylated targets. Mol Cancer Ther. 6:1089–1098. 2007. View Article : Google Scholar : PubMed/NCBI
19.	Schmittgen TD and Livak KJ: Analyzing real-time PCR data by the comparative CT method. Nat Protoc. 3:1101–1108. 2008. View Article : Google Scholar : PubMed/NCBI
20.	Zhang JJ, Zhang L, Zhou K, Ye X, Liu C, Zhang L, Kang J and Cai C: Analysis of global DNA methylation by hydrophilic interaction ultra high-pressure liquid chromatography tandem mass spectrometry. Anal Biochem. 413:164–170. 2011. View Article : Google Scholar : PubMed/NCBI
21.	Pogribny I, Yi P and James SJ: A sensitive new method for rapid detection of abnormal methylation patterns in global DNA and within CpG islands. Biochem Biophys Res Commun. 262:624–628. 1999. View Article : Google Scholar : PubMed/NCBI
22.	Kim TD, Shin S, Berry WL, Oh S and Janknecht R: The JMJDA demethylase regulates apoptosis and proliferation in colon cancer cells. J Cell Biochem. 113:1368–1376. 2012. View Article : Google Scholar : PubMed/NCBI
23.	Stewart MD, Li J and Wong J: Relationship between histone H3 lysine 9 methylation, transcription repression, and heterochromatin protein 1 recruitment. Mol Cell Biol. 25:2525–2538. 2005. View Article : Google Scholar : PubMed/NCBI
24.	Shi Y, Lan F, Matson C, Mulligan P, Whetstine JR, Cole PA, Casero RA and Shi Y: Histone demethylation mediated by the nuclear amine oxidase homolog LSD1. Cell. 119:941–953. 2004. View Article : Google Scholar : PubMed/NCBI
25.	Hileti D, Panayiotidis P and Hoffbrand AV: Iron chelators induce apoptosis in proliferating cells. Br J Haematol. 89:181–187. 1995. View Article : Google Scholar : PubMed/NCBI
26.	Pan YJ, Hopkins RG and Loo G: Increased GADD153 gene expression during iron chelation-induced apoptosis in Jurkat T-lymphocytes. Biochim Biophys Acta. 1691:41–50. 2004. View Article : Google Scholar : PubMed/NCBI
27.	So EY, Ausman M, Saeki T and Ouchi T: Phosphorylation of SMC1 by ATR is required for desferrioxamine (DFO)-induced apoptosis. Cell Death Dis. 2:e1282011. View Article : Google Scholar : PubMed/NCBI
28.	Saletta F, Suryo Rahmanto Y, Siafakas AR and Richardson DR: Cellular iron depletion and the mechanisms involved in the iron-dependent regulation of the growth arrest and DNA damage family of genes. J Biol Chem. 286:35396–35406. 2011. View Article : Google Scholar : PubMed/NCBI
29.	Escoubet-Lozach L, Lin IL, Jensen-Pergakes K, Brady HA, Gandhi AK, Schafer PH, Muller GW, Worland PJ, Chan KW and Verhelle D: Pomalidomide and lenalidomide induce p21WAF-1 expression in both lymphoma and multiple myeloma through a LSD1-mediated epigenetic mechanism. Cancer Res. 69:7347–7356. 2009. View Article : Google Scholar : PubMed/NCBI
30.	Siegel R, Naishadham D and Jemal A: Cancer statistics, 2012. CA Cancer J Clin. 62:10–29. 2012. View Article : Google Scholar
31.	Jemal A, Bray F, Center MM, Ferlay J, Ward E and Forman D: Global cancer statistics. CA Cancer J Clin. 61:69–90. 2011. View Article : Google Scholar
32.	Huang X: Does iron have a role in breast cancer? Lancet Oncol. 9:803–807. 2008. View Article : Google Scholar : PubMed/NCBI
33.	Pinnix ZK, Miller LD, Wang W, D’Agostino R Jr, Kute T, Willngham MC, Hatcher H, Tesfay L, Sui G, Di X, Torti SV and Torti FM: Ferroportin and iron regulation in breast cancer progression and prognosis. Sci Transl Med. 2:43ra562010. View Article : Google Scholar : PubMed/NCBI
34.	Shpyleva SI, Tryndyak VP, Kovalchuk O, Starlard-Davenport A, Chekhun VF, Beland FA and Pogribny IP: Role of ferritin alterations in human breast cancer cells. Breast Cancer Res Treat. 126:63–71. 2011. View Article : Google Scholar : PubMed/NCBI
35.	Hoke EM, Maylock CA and Shacter E: Desferal inhibits breast tumor growth and does not interfere with the tumoricidal activity of doxorubicin. Free Radic Biol Med. 39:403–411. 2005. View Article : Google Scholar : PubMed/NCBI
36.	Whitnall M, Howard J, Ponka P and Richardson DR: A class of iron chelators with a wide spectrum of potent antitumor activity that overcomes resistance to chemotherapeutics. Proc Natl Acad Sci USA. 103:14901–14906. 2006. View Article : Google Scholar : PubMed/NCBI
37.	Chen Z, Zhang D, Yue F, Zheng M, Kovacevic Z and Richardson DR: The iron chelators Dp44mT and DFO inhibit TGF-β-induced epithelial-mesenchymal transition via up-regulation of N-Myc downstream-regulated gene-1 (NDRG1). J Biol Chem. 287:17016–17028. 2012.PubMed/NCBI
38.	Pollard PJ, Loenarz C, Mole DR, McDonough MA, Gleadle JM, Schofield CJ and Ratcliffe PJ: Regulation of Jumonji-domain-containing histone demethylases by hypoxia-inducible factor (HIF)-1α. Biochem J. 416:387–394. 2008.
39.	Culhane JC and Cole PA: LSD1 and the chemistry of histone demethylation. Curr Opin Chem Biol. 11:561–568. 2007. View Article : Google Scholar : PubMed/NCBI
40.	Luka Z, Moss F, Loukachevitch LV, Bornhop DJ and Wagner C: Histone demethylase LSD1 is a folate-binding protein. Biochemistry. 50:4750–4756. 2011. View Article : Google Scholar : PubMed/NCBI
41.	Black JC, Allen A, Van Rechem C, Forbes E, Longworth M, Tschöp K, Rinehart C, Quiton J, Walsh R, Smallwood A, Dyson NJ and Whetstine JR: Conserved antagonism between JMJD2A/KDM4A and HP1γ during cell cycle progression. Mol Cell. 40:736–748. 2010.PubMed/NCBI
42.	Mallette FA, Mattiroli F, Cui G, Young LC, Hendzel MJ, Mer G, Sixma TK and Richard S: RNF8- and RNF168-dependent degradation of KDM4A/JMJD2A triggers 53BP1 recruitment to DNA damage sites. EMBO J. 31:1865–1878. 2012. View Article : Google Scholar : PubMed/NCBI
43.	Kim TD, Oh S, Shin S and Janknecht R: Regulation of tumor suppressor p53 and HCT116 cell physiology by histone demethylase JMJD2D/KDM4D. PLoS One. 7:e346182012. View Article : Google Scholar : PubMed/NCBI
44.	Binda O, LeRoy G, Bua DJ, Garcia BA, Gozani O and Richard S: Trimethylation of histone H3 lysine 4 impairs methylation of histone H3 lysine 9. Regulation of lysine methyltransferases by physical interaction with their substrates. Epigenetics. 5:767–775. 2010. View Article : Google Scholar : PubMed/NCBI
45.	Kovacevic Z, Sivagurunathan S, Mangs H, Chikhani S, Zhang D and Richardson DR: The metastasis suppressor, N-myc downstream regulated gene 1 (NDRG1), upregulates p21 via p53-independent mechanisms. Carcinogenesis. 32:732–740. 2011. View Article : Google Scholar : PubMed/NCBI
46.	Tryndyak VP, Beland FA and Pogribny IP: E-cadherin transcriptional down-regulation by epigenetic and microRNA-200 family alterations is related to mesenchymal and drug-resistant phenotypes in human breast cancer cells. Int J Cancer. 126:2575–2583. 2010.PubMed/NCBI