Differential effects of garcinol and curcumin on histone and p53 modifications in tumour cells

BMC Cancer

Hilary M Collins³  Magdy K Abdelghany¹  Marie Messmer³  Baigong Yue³  Sian E Deeves³  Karin B Kindle³  Kempegowda Mantelingu²  Akhmed Aslam³  G Sebastiaan Winkler³  Tapas K Kundu²  David M Heery³ 

[1] Present address: Department of Pathology, Faculty of Medicine, Suez Canal University, Ismailia, Egypt

[2] Transcription and Disease Laboratory, Jawaharlal Nehru Centre for Advanced Scientific Research, 560064, Bangalore, Karnataka, India

[3] Gene Regulation Group, Centre for Biomolecular Sciences, School of Pharmacy, University of Nottingham, University Park, NG7 2RD, Nottingham, United Kingdom

关键词: TIP60;    SUV420H2;    H4K20Me3;    Post-translational modifications;    p53;    Histones;    HAT inhibitor;    Acetyltransferase;    Curcumin;    Garcinol;   

Others  :  1079953
DOI  :  10.1186/1471-2407-13-37

 received in 2012-04-24, accepted in 2013-01-25,  发布年份 2013

【 摘 要 】

Background

Post-translational modifications (PTMs) of histones and other proteins are perturbed in tumours. For example, reduced levels of acetylated H4K16 and trimethylated H4K20 are associated with high tumour grade and poor survival in breast cancer. Drug-like molecules that can reprogram selected histone PTMs in tumour cells are therefore of interest as potential cancer chemopreventive agents. In this study we assessed the effects of the phytocompounds garcinol and curcumin on histone and p53 modification in cancer cells, focussing on the breast tumour cell line MCF7.

Methods

Cell viability/proliferation assays, cell cycle analysis by flow cytometry, immunodetection of specific histone and p53 acetylation marks, western blotting, siRNA and RT-qPCR.

Results

Although treatment with curcumin, garcinol or the garcinol derivative LTK-14 hampered MCF7 cell proliferation, differential effects of these compounds on histone modifications were observed. Garcinol treatment resulted in a strong reduction in H3K18 acetylation, which is required for S phase progression. Similar effects of garcinol on H3K18 acetylation were observed in the osteosarcoma cells lines U2OS and SaOS2. In contrast, global levels of acetylated H4K16 and trimethylated H4K20 in MCF7 cells were elevated after garcinol treatment. This was accompanied by upregulation of DNA damage signalling markers such as γH2A.X, H3K56Ac, p53 and TIP60. In contrast, exposure of MCF7 cells to curcumin resulted in increased global levels of acetylated H3K18 and H4K16, and was less effective in inducing DNA damage markers. In addition to its effects on histone modifications, garcinol was found to block CBP/p300-mediated acetylation of the C-terminal activation domain of p53, but resulted in enhanced acetylation of p53K120, and accumulation of p53 in the cytoplasmic compartment. Finally, we show that the elevation of H4K20Me3 levels by garcinol correlated with increased expression of SUV420H2, and was prevented by siRNA targeting of SUV420H2.

Conclusion

In summary, although garcinol and curcumin can both inhibit histone acetyltransferase activities, our results show that these compounds have differential effects on cancer cells in culture. Garcinol treatment alters expression of chromatin modifying enzymes in MCF7 cells, resulting in reprogramming of key histone and p53 PTMs and growth arrest, underscoring its potential as a cancer chemopreventive agent.

【 授权许可】

   
2013 Collins et al; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

【 参考文献 】

• [1]Kouzarides T: Chromatin modifications and their function. Cell 2007, 128:693-705.
• [2]Jenuwein T, Allis CD: Translating the histone code. Science 2001, 293:1074-1080.
• [3]Tjeertes JV, Miller KM, Jackson SP: Screen for DNA-damage-responsive histone modifications identifies H3K9Ac and H3K56Ac in human cells. EMBO J 2009, 28:1878-1889.
• [4]Taverna SD, Li H, Ruthenburg AJ, Allis CD, Patel DJ: How chromatin-binding modules interpret histone modifications: lessons from professional pocket pickers. Nat Struct Mol Biol 2007, 14:1025-1040.
• [5]van Attikum H, Gasser SM: Crosstalk between histone modifications during the DNA damage response. Trends Cell Biol 2009, 19:207-217.
• [6]Fraga MF, Ballestar E, Villar-Garea A, Boix-Chornet M, Espada J, Schotta G, Bonaldi T, Haydon C, Ropero S, Petrie K: Loss of acetylation at Lys16 and trimethylation at Lys20 of histone H4 is a common hallmark of human cancer. Nat Genet 2005, 37:391-400.
• [7]Elsheikh SE, Green AR, Rakha EA, Powe DG, Ahmed RA, Collins HM, Soria D, Garibaldi JM, Paish CE, Ammar AA: Global histone modifications in breast cancer correlate with tumor phenotypes, prognostic factors, and patient outcome. Cancer Res 2009, 69:3802-3809.
• [8]Seligson DB, Horvath S, Shi T, Yu H, Tze S, Grunstein M, Kurdistani SK: Global histone modification patterns predict risk of prostate cancer recurrence. Nature 2005, 435:1262-1266.
• [9]Barlesi F, Giaccone G, Gallegos-Ruiz MI, Loundou A, Span SW, Lefesvre P, Kruyt FA, Rodriguez JA: Global histone modifications predict prognosis of resected non small-cell lung cancer. J Clin Oncol 2007, 25:4358-4364.
• [10]Park YS, Jin MY, Kim YJ, Yook JH, Kim BS, Jang SJ: The global histone modification pattern correlates with cancer recurrence and overall survival in gastric adenocarcinoma. Ann Surg Oncol 2008, 15:1968-1976.
• [11]Sharma GG, So S, Gupta A, Kumar R, Cayrou C, Avvakumov N, Bhadra U, Pandita RK, Porteus MH, Chen DJ: MOF and histone H4 acetylation at lysine 16 are critical for DNA Damage Response and DSB Repair. Mol Cell Biol 2010, 30:3582-3585.
• [12]Sanders SL, Portoso M, Mata J, Bahler J, Allshire RC, Kouzarides T: Methylation of histone H4 lysine 20 controls recruitment of Crb2 to sites of DNA damage. Cell 2004, 119:603-614.
• [13]Heery DM, Fischer PM: Pharmacological targeting of lysine acetyltransferases in human disease: a progress report. Drug Discov Today 2007, 12:88-99.
• [14]Hodawadekar SC, Marmorstein R: Chemistry of acetyl transfer by histone modifying enzymes: structure, mechanism and implications for effector design. Oncogene 2007, 26:5528-5540.
• [15]Arif M, Pradhan SK, Thanuja GR, Vedamurthy BM, Agrawal S, Dasgupta D, Kundu TK: Mechanism of p300 specific histone acetyltransferase inhibition by small molecules. J Med Chem 2009, 52:267-277.
• [16]Balasubramanyam K, Varier RA, Altaf M, Swaminathan V, Siddappa NB, Ranga U, Kundu TK: Curcumin, a novel p300/CREB-binding protein-specific inhibitor of acetyltransferase, represses the acetylation of histone/nonhistone proteins and histone acetyltransferase-dependent chromatin transcription. J Biol Chem 2004, 279:51163-51171.
• [17]Balasubramanyam K, Altaf M, Varier RA, Swaminathan V, Ravindran A, Sadhale PP, Kundu TK: Polyisoprenylated benzophenone, garcinol, a natural histone acetyltransferase inhibitor, represses chromatin transcription and alters global gene expression. J Biol Chem 2004, 279:33716-33726.
• [18]Mantelingu K, Reddy BA, Swaminathan V, Kishore AH, Siddappa NB, Kumar GV, Nagashankar G, Natesh N, Roy S, Sadhale PP: Specific inhibition of p300-HAT alters global gene expression and represses HIV replication. Chem Biol 2007, 14:645-657.
• [19]Shechter D, Dormann HL, Allis CD, Hake SB: Extraction, purification and analysis of histones. Nat Protoc 2007, 2:1445-1457.
• [20]Aslam A, Mittal S, Koch F, Andrau JC, Winkler GS: The Ccr4-not deadenylase subunits CNOT7 and CNOT8 have overlapping roles and modulate cell proliferation. Mol Biol Cell 2009, 20:3840-3850.
• [21]Schneider CA, Rasband WS, Eliceiri KW: NIH Image to ImageJ: 25 years of image analysis. Nat Methods 2012, 9:671-675.
• [22]Horwitz GA, Zhang K, McBrian MA, Grunstein M, Kurdistani SK, Berk AJ: Adenovirus small e1a alters global patterns of histone modification. Science 2008, 321:1084-1085.
• [23]Ewald B, Sampath D, Plunkett W: H2AX phosphorylation marks gemcitabine-induced stalled replication forks and their collapse upon S-phase checkpoint abrogation. Mol Cancer Ther 2007, 6:1239-1248.
• [24]Gu W, Roeder RG: Activation of p53 sequence-specific DNA binding by acetylation of the p53 C-terminal domain. Cell 1997, 90:595-606.
• [25]Sykes SM, Mellert HS, Holbert MA, Li K, Marmorstein R, Lane WS, McMahon SB: Acetylation of the p53 DNA-binding domain regulates apoptosis induction. Mol Cell 2006, 24:841-851.
• [26]Sykes SM, Stanek TJ, Frank A, Murphy ME, McMahon SB: Acetylation of the DNA binding domain regulates transcription-independent apoptosis by p53. J Biol Chem 2009, 284:20197-20205.
• [27]Tang Y, Luo J, Zhang W, Gu W: Tip60-dependent acetylation of p53 modulates the decision between cell-cycle arrest and apoptosis. Mol Cell 2006, 24:827-839.
• [28]Pesavento JJ, Yang H, Kelleher NL, Mizzen CA: Certain and progressive methylation of histone H4 at lysine 20 during the cell cycle. Mol Cell Biol 2008, 28:468-486.
• [29]Tsang LW, Hu N, Underhill DA: Comparative analyses of SUV420H1 isoforms and SUV420H2 reveal differences in their cellular localization and effects on myogenic differentiation. PLoS One 2010, 5:e14447.
• [30]Yoshida K, Tanaka T, Hirose Y, Yamaguchi F, Kohno H, Toida M, Hara A, Sugie S, Shibata T, Mori H: Dietary garcinol inhibits 4-nitroquinoline 1-oxide-induced tongue carcinogenesis in rats. Cancer Lett 2005, 221:29-39.
• [31]Yamaguchi F, Ariga T, Yoshimura Y, Nakazawa H: Antioxidative and anti-glycation activity of garcinol from Garcinia indica fruit rind. J Agric Food Chem 2000, 48:180-185.
• [32]Ahmad A, Wang Z, Ali R, Maitah MY, Kong D, Banerjee S, Padhye S, Sarkar FH: Apoptosis-inducing effect of garcinol is mediated by NF-kappaB signaling in breast cancer cells. J Cell Biochem 2010, 109:1134-1141.
• [33]Sahu RP, Batra S, Srivastava SK: Activation of ATM/Chk1 by curcumin causes cell cycle arrest and apoptosis in human pancreatic cancer cells. Br J Cancer 2009, 100:1425-1433.
• [34]Kim SJ, Son TG, Park HR, Park M, Kim MS, Kim HS, Chung HY, Mattson MP, Lee J: Curcumin stimulates proliferation of embryonic neural progenitor cells and neurogenesis in the adult hippocampus. J Biol Chem 2008, 283:14497-14505.
• [35]Kim JH, Park SH, Nam SW, Kwon HJ, Kim BW, Kim WJ, Choi YH: Curcumin stimulates proliferation, stemness acting signals and migration of 3T3-L1 preadipocytes. Int J Mol Med 2011, 28:429-435.
• [36]Troke PJ, Kindle KB, Collins HM, Heery DM: MOZ fusion proteins in acute myeloid leukaemia. Biochem Soc Symp 2006, 23-39.
• [37]Jin Q, Yu LR, Wang L, Zhang Z, Kasper LH, Lee JE, Wang C, Brindle PK, Dent SY, Ge K: Distinct roles of GCN5/PCAF-mediated H3K9ac and CBP/p300-mediated H3K18/27ac in nuclear receptor transactivation. EMBO J 2011, 30:249-262.
• [38]Ferrari R, Pellegrini M, Horwitz GA, Xie W, Berk AJ, Kurdistani SK: Epigenetic reprogramming by adenovirus e1a. Science 2008, 321:1086-1088.
• [39]Taipale M, Rea S, Richter K, Vilar A, Lichter P, Imhof A, Akhtar A: hMOF histone acetyltransferase is required for histone H4 lysine 16 acetylation in mammalian cells. Mol Cell Biol 2005, 25:6798-6810.
• [40]Smith ER, Cayrou C, Huang R, Lane WS, Cote J, Lucchesi JC: A human protein complex homologous to the Drosophila MSL complex is responsible for the majority of histone H4 acetylation at lysine 16. Mol Cell Biol 2005, 25:9175-9188.
• [41]Kusch T, Florens L, Macdonald WH, Swanson SK, Glaser RL, Yates JR 3rd, Abmayr SM, Washburn MP, Workman JL: Acetylation by Tip60 is required for selective histone variant exchange at DNA lesions. Science 2004, 306:2084-2087.
• [42]Ikura T, Tashiro S, Kakino A, Shima H, Jacob N, Amunugama R, Yoder K, Izumi S, Kuraoka I, Tanaka K: DNA damage-dependent acetylation and ubiquitination of H2AX enhances chromatin dynamics. Mol Cell Biol 2007, 27:7028-7040.
• [43]Kozak ML, Chavez A, Dang W, Berger SL, Ashok A, Guo X, Johnson FB: Inactivation of the Sas2 histone acetyltransferase delays senescence driven by telomere dysfunction. EMBO J 2010, 29:158-170.
• [44]Shogren-Knaak M, Ishii H, Sun JM, Pazin MJ, Davie JR, Peterson CL: Histone H4-K16 acetylation controls chromatin structure and protein interactions. Science 2006, 311:844-847.
• [45]Lu X, Simon MD, Chodaparambil JV, Hansen JC, Shokat KM, Luger K: The effect of H3K79 dimethylation and H4K20 trimethylation on nucleosome and chromatin structure. Nat Struct Mol Biol 2008, 15:1122-1124.
• [46]Kapoor-Vazirani P, Kagey JD, Vertino PM: SUV420H2-mediated H4K20 trimethylation enforces RNA polymerase II promoter-proximal pausing by blocking hMOF-dependent H4K16 acetylation. Mol Cell Biol 2011, 31:1594-1609.