Reversibility of membrane N-glycome of HeLa cells upon treatment with epigenetic inhibitors

PLoS One. 2013;8(1):e54672. doi: 10.1371/journal.pone.0054672. Epub 2013 Jan 15.

Abstract

Glycans are essential regulators of protein function and are now in the focus of research in many physiological and pathophysiological processes. There are numerous modes of regulating their biosynthesis, including epigenetic mechanisms implicated in the expression of glyco-genes. Since N-glycans located at the cell membrane define intercellular communication as well as a cellular response to a given environment, we developed a method to preferentially analyze this fraction of glycans. The method is based on incorporation of living cells into polyacrylamide gels, partial denaturation of membrane proteins with 3 M urea and subsequent release of N-glycans with PNGase F followed by HPLC analysis. Using this newly developed method, we revealed multiple effects of epigenetic inhibitors Trichostatin A, sodium butyrate and zebularine on the composition of N-glycans in human cells. The induced changes were found to be reversible after inhibitor removal. Given that many epigenetic inhibitors are currently explored as a therapeutic strategy in treatment of cancer, wherein surface glycans play an important role, the presented work contributes to our understanding of their efficiency in altering the N-glycan profile of cancer cells in culture.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Butyrates / pharmacology
  • Cell Membrane / drug effects*
  • Cell Membrane / metabolism*
  • Cytidine / analogs & derivatives
  • Cytidine / pharmacology
  • Epigenesis, Genetic / drug effects
  • HeLa Cells
  • Histone Deacetylase Inhibitors / pharmacology
  • Humans
  • Hydroxamic Acids / pharmacology
  • Polysaccharides / metabolism*

Substances

  • Butyrates
  • Histone Deacetylase Inhibitors
  • Hydroxamic Acids
  • Polysaccharides
  • trichostatin A
  • Cytidine
  • pyrimidin-2-one beta-ribofuranoside

Grant support

This work was supported by the Croatian Ministry of Science, Education and Sport grants #309-0061194-2023 (to GL), #119-1191196-1224 (to VZ) and #098-0982464-2513 (to MHB); by the European Commission GlycoBioM (contract #259869) and HighGlycan (contract #278535) grants and by AUF PSCI Grant (Agence universitaire de la Francophonie). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.