title: Reversibility of membrane N-glycome of HeLa cells upon treatment with epigenetic inhibitors.
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.
authors: Faculty of Science, University of Zagreb, Zagreb, Croatia.
authors: Tomislav
authors: T
authors: Horvat
authors: Martina
authors: M
authors: Deželjin
authors: Irma
authors: I
authors: Redžić
authors: Darko
authors: D
authors: Barišić
authors: Maja
authors: M
authors: Herak Bosnar
authors: Gordan
authors: G
authors: Lauc
authors: Vlatka
authors: V
authors: Zoldoš
mesh_terms: D002087:Butyrates / Q000494:pharmacology; D002462:Cell Membrane / Q000187:drug effects* / Q000378:metabolism*; D003562:Cytidine / Q000031:analogs & derivatives / Q000494:pharmacology; D044127:Epigenesis, Genetic / Q000187:drug effects; D006367:HeLa Cells; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D011134:Polysaccharides / Q000378:metabolism*
chemical_list: D002087:Butyrates; D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids; D011134:Polysaccharides; C012589:trichostatin A; D003562:Cytidine; C009131:pyrimidin-2-one beta-ribofuranoside