1 Scientific RepoRts | 6:27965 | DOI: 10.1038/srep27965 www.nature.com/scientificreports Intrinsic hepatocyte dediferentiation is accompanied by upregulation of mesenchymal markers, protein sialylation and core alpha 1,6 linked fucosylation Anand Mehta 1 , Mary Ann Comunale 1 , siddhartha Rawat 2 , Jessica C. Casciano 2 , Jason Lamontagne 3 , Harmin Herrera 3 , Aarti Ramanathan 3 , Lucy Betesh 1 , Mengjun Wang 1 , pamela Norton 1 , Laura F. steel 4 & Michael J. Bouchard 5 Alterations in N-linked glycosylation have long been associated with cancer but for the most part, the reasons why have remained poorly understood. Here we show that increased core fucosylation is associated with de-diferentiation of primary hepatocytes and with the appearance of markers indicative of a transition of cells from an epithelial to a mesenchymal state. this increase in core fucosylation was associated with increased levels of two enzymes involved in α-1,6 linked fucosylation, GDP-mannose 4, 6-dehydratase (Gmds) and to a lesser extent fucosyltransferase 8 (Fut8). In addition, the activation of cancer-associated cellular signaling pathways in primary rat hepatocytes can increase core fucosylation and induce additional glycoform alterations on hepatocyte proteins. Specifcally, we show that increased levels of protein sialylation and α-1,6-linked core fucosylation are observed following activation of the β-catenin pathway. Activation of the Akt signaling pathway or induction of hypoxia also results in increased levels of fucosylation and sialylation. We believe that this knowledge will help in the better understanding of the genetic factors associated with altered glycosylation and may allow for the development of more clinically relevant biomarkers. Despite advances in medical technology, the 5-year survival rate for hepatocellular carcinoma (HCC) is only 8–13%, likely due to the fact that the majority of patients with HCC are diagnosed at advanced stages 1 . HCC is one of the most common solid malignancies worldwide, and the incidence in the United States is increasing 2 . In this setting of a signifcant increase in the number of patients with HCC, early detection and treatment are vital to improve the otherwise dismal outcome of this disease 3,4 . Te urgent need for biomarkers for the early detection of HCC is widely recognized, and many strategies are being pursued to achieve this goal. In our previous work, we identifed increased levels of core and outer-arm fucosylation of a large number of serum and liver tissue proteins as being associated with HCC 5–8 and similar increases in fucosylation have been seen in other cancers 9–17 . In an efort to further understand why these changes in glycosylation occur we have used cultured primary rat hepatocytes (PRH) to study the consequence of an epithelial to mesenchymal transition (EMT) on N-linked glycosylation. In addition, we provide evidence that activation of specifc cancer-associated pathways associated 1 Drexel University College of Medicine, Department of Microbiology and Immunology, 245 N. 15th Street, Philadelphia, PA 19102, USA. 2 Graduate School of Biomedical Sciences and Professional Studies, Drexel University College of Medicine, Molecular and Cellular Biology and Genetics Graduate Program, 245 North 15th Street, Philadelphia, PA 19102, USA. 3 Graduate School of Biomedical Sciences and Professional Studies, Drexel University College of Medicine, Microbiology and Immunology Graduate Program, 2900 Queen Lane, Philadelphia, PA 19129, USA. 4 Drexel University college of Medicine, Department of Microbiology and immunology, institute for Molecular Medicine and Infectious Disease, 245 North 15th Street, Philadelphia, PA 19102, USA. 5 Drexel University college of Medicine, Department of Biochemistry and Molecular Biology, 245 N. 15th Street, Philadelphia, PA 19102, USA. Correspondence and requests for materials should be addressed to A.M. (email: anand.mehta@drexelmed.edu) received: 29 September 2015 Accepted: 25 May 2016 Published: 22 June 2016 opeN