The International Journal of Biochemistry & Cell Biology 79 (2016) 69–79 Contents lists available at ScienceDirect The International Journal of Biochemistry & Cell Biology journal homepage: www.elsevier.com/locate/biocel Inhibiting post-translational core fucosylation prevents vascular calcification in the model of uremia Xinyu Wen a,b,1 , Anqi Liu a,b,1 , Changqing Yu b , Lingyu Wang b , Mengying Zhou b , Nan Wang b , Ming Fang b , Weidong Wang b , Hongli Lin b,∗ a Graduate School of Dalian Medical University, Dalian, China b Department of Nephrology, Liaoning Translational Medicine Center of Nephrology, The First Affiliated Hospital of Dalian Medical University, Dalian, China a r t i c l e i n f o Article history: Received 1 December 2015 Received in revised form 20 June 2016 Accepted 9 August 2016 Available online 10 August 2016 Keywords: -1,6-fucosyltransferase Vascular calcification TGF-R a b s t r a c t Vascular calcification (VC) is an independent risk factor for cardiovascular disease and mortality in ure- mia. Post-translational core fucosylation is implicated in a number of pathological processes. First, we investigated the role of core fucosylation and key TGF-1 pathway receptors in calcified arteries in vivo. To determine whether blocking core fucosylation effectively inhibited VC and TGF-/Smad signaling pathway, we established an in vitro model of phosphate-induced calcification in rat vascular smooth muscle cells (VSMCs) to assess the role of core fucosylation in VC. Core fucose could be detected at markedly higher levels in calcified VSMCs than control cells. Fut8 (-1,6 fucosyltransferase), the only enzyme responsible for core fucosylation in humans, was significantly upregulated by high phosphate. Exposed to high phosphate media and blocking core fucosylation in VSMCs by knocking down Fut8 using a siRNA markedly reduced calcium and phosphorus deposition and Cbf1 expression (osteoblast-specific transcription factor), and increased -Sma expression (smooth muscle cell marker). Fut8 siRNA signif- icantly inhibited TGF-/Smad2/3 signaling activation in VSMCs cultured in high phosphate media. In conclusion, this study provides evidence to suggest core fucosylation plays a major role in the process of VC and appropriate blockade of core fucosylation may represent a potential therapeutic strategy for treating VC in end-stage renal disease. © 2016 Published by Elsevier Ltd. 1. Introduction Vascular calcification (VC) is a common complication and the major cause of cardiovascular disease in patients with end-stage renal disease (Karohl et al., 2011). The prevalence of VC increases during the progression of chronic kidney disease (CKD), ranging from 40% among patients with stage 3 CKD to 80–99% in patients with end-stage renal disease on dialysis (Garland et al., 2008; Sigrist et al., 2006; Adeney et al., 2009; Chertow et al., 2002). However, the precise molecular mechanisms underlying VC still need to be clarified. VC is no longer regarded as a passive process and is con- sidered an actively-regulated and complex process that is not yet Abbreviation: VSMCs, vascular smooth muscle cells; CKD, chronic kidney dis- ease; Fut8, -1,6 fucosyltransferase; cbf1, core-binding factor subunit 1 ; -SMA, -smooth muscle actin; TGF1, transforming growth factor-1; LCA-FITC, fluores- cent L. culinaris agglutinin–fluorescein complex. ∗ Corresponding author at: Department of Nephrology, The First Affiliated Hospi- tal of Dalian Medical University, No. 222 Zhongshan Road, Dalian 116011, China. E-mail address: hllin@dlmedu.edu.cn (H. Lin). 1 These authors contributed equally to this work and share the first authorship. completely understood. During VC, the specific, indispensable tran- scriptional regulator of osteoblastic differentiation core-binding factor subunit 1 (Cbf1) is upregulated, while expression of the VSMC marker -smooth muscle actin (-SMA) decreases (Giachelli et al., 2005; Hruska et al., 2005). A number of cytokines and signaling pathways have been demonstrated to stimulate the occurrence and development of VC, including TNF-, osteonectin, osteocalcin, the BMP-2 signaling pathway, transforming growth factor- (TGF-) signaling path- way (Yetkin and Waltenberger, 2009; Wang et al., 2013) and the Wnt/-catenin/OPG/RANKL/RANK axis (Evrard et al., 2015). These studies have provided important data regarding the mechanisms that underlie VC and helped to infer the process that occur during the progression of VC. Most of the above-mentioned studies have explored the roles of key proteins in VC by altering the expression of these genes or proteins. In fact, in addition to gene and pro- tein expression levels, post-translational modifications of proteins can also have a major effect on protein function. Data increasingly indicates that post-translational modifications directly and defini- tively regulate protein function in a range of pathophysiological processes, and in some cases, this regulation is independent of the http://dx.doi.org/10.1016/j.biocel.2016.08.015 1357-2725/© 2016 Published by Elsevier Ltd.