Prenylated chalcone xanthohumol associates with histones in breast cancer cells–a novel target identified by a monoclonal antibody
Article first published online: 24 SEP 2012
© 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Molecular Nutrition & Food Research
Volume 56, Issue 11, pages 1688–1696, November 2012
How to Cite
Wyns, C., van Steendam, K., Vanhoecke, B., Deforce, D., Bracke, M. and Heyerick, A. (2012), Prenylated chalcone xanthohumol associates with histones in breast cancer cells–a novel target identified by a monoclonal antibody. Mol. Nutr. Food Res., 56: 1688–1696. doi: 10.1002/mnfr.201200030
- Issue published online: 24 OCT 2012
- Article first published online: 24 SEP 2012
- Manuscript Accepted: 10 AUG 2012
- Manuscript Revised: 31 JUL 2012
- Manuscript Received: 16 JAN 2012
- Institute for the Promotion of Innovation through Science
- Technology in Flanders
- Concerted Research Initiative of the Ghent University. Grant Number: 01G013A7
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 μM 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 μM 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.
In summary, we did successfully apply a mAB against XN in immunocytochemistry and precipitation with highly unexpected results.