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Prenylated chalcone xanthohumol associates with histones in breast cancer cells: a novel target identified by a monoclonal antibody

Ciska Wyns (UGent) , Katleen Van Steendam (UGent) , Barbara Vanhoecke (UGent) , Dieter Deforce (UGent) , Marc Bracke (UGent) and Arne Heyerick (UGent)
(2012) MOLECULAR NUTRITION & FOOD RESEARCH. 56(11). p.1688-1696
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Abstract
Scope : The intracellular fate of xanthohumol (XN) from hops is an underexplored field in the research for the molecular mechanisms causing its wide range of effects in chemoprevention and gene expression involved in hepatic metabolism. Methods and results : We aimed to elucidate possible targets for binding of XN in a human mammary carcinoma cell line (MCF-7/6), using a mAB. We investigated the overall solubility and stability of XN in growth medium and the cellular uptake and distribution of XN in MCF-7/6 cells using an optimized immunocytochemistry technique. After incubation of MCF-7/6 cells, with 10 mu M XN for 0.5 h up to 6 h, we observed primarily a granular nuclear staining, which intensified with increasing exposure times. Immunoprecipitation of cell lysates (treated with 10 mu M XN for 2 h) revealed binding of XN to a fraction of proteins with a molecular weight below 20 kDa. Further analysis of the protein mixture via LC-MS/MS (Q-TOF) resulted in the identification of specific members of the histone family, i.e. histone H2A, H2B, and H4. The identity of histone H2A was confirmed using immunodetection with a specific anti-histone H2A antibody. Conclusion : In summary, we did successfully apply a mAB against XN in immunocytochemistry and precipitation with highly unexpected results.
Keywords
Immunocytochemistry, Histones, Immunoprecipitation, MCF-7, 6, Xanthohumol, HOPS HUMULUS-LUPULUS, HUMAN LIVER-MICROSOMES, NF-KAPPA-B, QUINONE REDUCTASE, L., BEER, PRENYLFLAVONOIDS, METABOLISM, ISOXANTHOHUMOL, CONSTITUENTS

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MLA
Wyns, Ciska, Katleen Van Steendam, Barbara Vanhoecke, et al. “Prenylated Chalcone Xanthohumol Associates with Histones in Breast Cancer Cells: a Novel Target Identified by a Monoclonal Antibody.” MOLECULAR NUTRITION & FOOD RESEARCH 56.11 (2012): 1688–1696. Print.
APA
Wyns, C., Van Steendam, K., Vanhoecke, B., Deforce, D., Bracke, M., & Heyerick, A. (2012). Prenylated chalcone xanthohumol associates with histones in breast cancer cells: a novel target identified by a monoclonal antibody. MOLECULAR NUTRITION & FOOD RESEARCH, 56(11), 1688–1696.
Chicago author-date
Wyns, Ciska, Katleen Van Steendam, Barbara Vanhoecke, Dieter Deforce, Marc Bracke, and Arne Heyerick. 2012. “Prenylated Chalcone Xanthohumol Associates with Histones in Breast Cancer Cells: a Novel Target Identified by a Monoclonal Antibody.” Molecular Nutrition & Food Research 56 (11): 1688–1696.
Chicago author-date (all authors)
Wyns, Ciska, Katleen Van Steendam, Barbara Vanhoecke, Dieter Deforce, Marc Bracke, and Arne Heyerick. 2012. “Prenylated Chalcone Xanthohumol Associates with Histones in Breast Cancer Cells: a Novel Target Identified by a Monoclonal Antibody.” Molecular Nutrition & Food Research 56 (11): 1688–1696.
Vancouver
1.
Wyns C, Van Steendam K, Vanhoecke B, Deforce D, Bracke M, Heyerick A. Prenylated chalcone xanthohumol associates with histones in breast cancer cells: a novel target identified by a monoclonal antibody. MOLECULAR NUTRITION & FOOD RESEARCH. 2012;56(11):1688–96.
IEEE
[1]
C. Wyns, K. Van Steendam, B. Vanhoecke, D. Deforce, M. Bracke, and A. Heyerick, “Prenylated chalcone xanthohumol associates with histones in breast cancer cells: a novel target identified by a monoclonal antibody,” MOLECULAR NUTRITION & FOOD RESEARCH, vol. 56, no. 11, pp. 1688–1696, 2012.
@article{3123722,
  abstract     = {Scope : The intracellular fate of xanthohumol (XN) from hops is an underexplored field in the research for the molecular mechanisms causing its wide range of effects in chemoprevention and gene expression involved in hepatic metabolism.
Methods and results : We aimed to elucidate possible targets for binding of XN in a human mammary carcinoma cell line (MCF-7/6), using a mAB. We investigated the overall solubility and stability of XN in growth medium and the cellular uptake and distribution of XN in MCF-7/6 cells using an optimized immunocytochemistry technique. After incubation of MCF-7/6 cells, with 10 mu M XN for 0.5 h up to 6 h, we observed primarily a granular nuclear staining, which intensified with increasing exposure times. Immunoprecipitation of cell lysates (treated with 10 mu M XN for 2 h) revealed binding of XN to a fraction of proteins with a molecular weight below 20 kDa. Further analysis of the protein mixture via LC-MS/MS (Q-TOF) resulted in the identification of specific members of the histone family, i.e. histone H2A, H2B, and H4. The identity of histone H2A was confirmed using immunodetection with a specific anti-histone H2A antibody.
Conclusion : In summary, we did successfully apply a mAB against XN in immunocytochemistry and precipitation with highly unexpected results.},
  author       = {Wyns, Ciska and Van Steendam, Katleen and Vanhoecke, Barbara and Deforce, Dieter and Bracke, Marc and Heyerick, Arne},
  issn         = {1613-4125},
  journal      = {MOLECULAR NUTRITION & FOOD RESEARCH},
  keywords     = {Immunocytochemistry,Histones,Immunoprecipitation,MCF-7,6,Xanthohumol,HOPS HUMULUS-LUPULUS,HUMAN LIVER-MICROSOMES,NF-KAPPA-B,QUINONE REDUCTASE,L.,BEER,PRENYLFLAVONOIDS,METABOLISM,ISOXANTHOHUMOL,CONSTITUENTS},
  language     = {eng},
  number       = {11},
  pages        = {1688--1696},
  title        = {Prenylated chalcone xanthohumol associates with histones in breast cancer cells: a novel target identified by a monoclonal antibody},
  url          = {http://dx.doi.org/10.1002/mnfr.201200030},
  volume       = {56},
  year         = {2012},
}

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