engleski

Epigenetic modulation of HeLa cell membrane N-glycome by epigenetic inhibitors and reversibility of inhibition effects in a drug-free environment

Deregulation of glycosylation occurs in a wide range of multifactorial diseases and aberrant glyco-phenotypes have been associated with specific pathophysiological states. Glycogenes are one of the groups of cancer-associated genes since changes in glycan structures are hallmarks of many cancers.Cancer-specific changes in glycan biosynthetic pathways are resulting from aberrant expressions of glycosyltransferases and glycosidases, which is often a result of epigenetic changes including DNA methylation and/or histone modifications (acetylation, methylation or phosphorylation). The aberrant gene expression pattern can be potentially restored using various epigenetic inhibitors. To test potential therapeutic usefulness of DNA methylation inhibitors, zebularine and 5- aza-2-deoxycytidine, and histone deacetylation inhibitors, trichostatin A and Na-butyrate, in inducing a reversal of undesired glyco-phenotypes, we developed an HPLC- based method for the analysis of glycan structures from HeLa cells embedded in polyacrylamide gels. In addition, we specifically investigated the preservation of altered glycan profiles over a prolonged period of time in a drug-free environment. Our results emphasize the importance of epigenetic control in the regulation of N-glycosylation, but also suggest the stability of complex biosynthetic pathways responsible for the establishment of glycan profiles in human cells in culture.

surface N-glycans; zebularine; 5-aza-2-deoxycytidine; Trichostatin A; HPLC analysis

nije evidentirano