Reversibility of Membrane N-Glycome of HeLa Cells upon Treatment with Epigenetic Inhibitors

Horvat, Tomislav; Deželjin, Martina; Redžić, Irma; Barišić, Darko; Herak Bosnar, Maja; Lauc, Gordan; Zoldoš, Vlatka (2013) Reversibility of Membrane N-Glycome of HeLa Cells upon Treatment with Epigenetic Inhibitors. PLoS ONE, 8 (1). e54672/1-e54672/9. ISSN 1932-6203

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Official URL: http://journals.plos.org/plosone/article?id=10.137...

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.

Item Type:

Article

Uncontrolled Keywords:

epigenetics; cell membrane N-glycome; zebularine; trichostatin A; HeLa cells; HPLC analysis

Subjects:

NATURAL SCIENCES > Biology

Divisions:

Division of Molecular Medicine

Projects:

Project title	Project leader	Project code	Project type
Dinamika kromatina i plastičnost genoma	[204780] Vlatka Zoldoš	119-1191196-1224	MZOS
Uloga gena nm23 u karcinomu pločastog epitela usne šupljine	[202030] Maja Herak-Bosnar	098-0982464-2513	MZOS

Depositing User:

Martina Deželjin

Date Deposited:

06 Jul 2015 12:24

URI:

http://fulir.irb.hr/id/eprint/2021

DOI:

10.1371/journal.pone.0054672