Blocking Posttranslational Core Fucosylation Ameliorates Rat Peritoneal Mesothelial Cell Epithelial-Mesenchymal Transition
- Author:
- No author available
- Journal Title:
- Chinese Medical Journal
- Issue:
- 18
- DOI:
- 10.4103/0366-6999.213963
- Key Word:
- α-1,6 Fucosyltransferase;Core Fucosylation;Epithelial-Mesenchymal Transition;Peritoneal Fibrosis
Abstract： Background:Core fucosylation (CF),catalyzed by α-1,6 fucosyltransferase (Fut8) in mammals,plays an important role in pathological processes through posttranslational modification of key signaling receptor proteins,including transforming growth factor (TGF)-β receptors and platelet-derived growth factor (PDGF) receptors.However,its effect on peritoneal fibrosis is unknown.Here,we investigated its influence on epithelial-mesenchymal transition (EMT) of rat peritoneal mesothelial cells (PMCs) in vitro induced by a high-glucose (HG) culture solution.Methods:Rat PMCs were first cultured in a HG (2.5％) culture solution to observe the CF expression level (fluorescein isothiocyanate-lens culinaris agglutinin),we next established a knockdown model of rat PMCs in vitro with Fut8 small interfering RNA (siRNA) to observe whether inhibiting CF decreases the messenger RNA (mRNA) expression and protein expression of Fut8 and reverses EMT status.Rat PMCs were randomly divided into control group,mock group (transfected with scrambled siRNA),Fut8 siRNA group,HG group,HG + mock group,and HG + Fut8 siRNA group.Finally,we examined the activation of TGF-β/Smad2/3 signaling and PDGF/extracellular signal-regulated kinase (ERK) signaling to observe the influence of CF on them.Results:CF,Fut8 mRNA,and protein expression were all significantly upregulated in HG-induced EMT model than those in the control rat PMCs (P ＜ 0.05).Fut8 siRNA successfully blocked CF of TGF-β receptors and PDGF receptors and attenuated the EMT status (E-cadherin and α-SMA and phenotypic changes) in HG-induced rat PMCs.In TGF-β/Smad2/3 signaling,Fut8 siRNA did not suppress the protein expression of TGF-3 receptors and Smad2/3;however,it significantly suppressed the phosphowlation of Smad2/3 (relative expression folds of HG + Fut8 group vs.HG group:7.6 ± 0.4 vs.15.1 ± 0.6,respectively,P ＜ 0.05).In PDGF/ERK signaling,Fut8 siRNA did not suppress the protein expression of PDGF receptors and ERK,but it significantly suppressed the phosphorylation of ERK (relative expression folds of HG + Fut8 group vs.HG group:8.7 ± 0.9 vs.15.6 ± 1.2,respectively,P ＜ 0.05).Blocking CF inactivated the activities of TGF-β and PDGF signaling pathways,and subsequently blocked EMT.Conclusions:These results demonstrate that CF contributes to rat PMC EMT.and that blocking it attenuates EMT.CF regulation is a potential therapeutic target of peritoneal fibrosis.
- [1]Wei Gu,Tomohiko Fukuda,Tomoya Isaji,etc.α1,6-Fucosylation regulates neurite formation via the activin/phospho-Smad2 pathway in PC12 cells: the implicated dual effects of Fut8 for TGF-β/activin-mediated signaling.[J].2013,27(10).
- [2]Gomes,K.B.,Marinho,M.A.S.,Otoni,A.,etc.Inflammation, neoangiogenesis and fibrosis in peritoneal dialysis[J].2013,421.
- [3]Hongli Lin,Dapeng Wang,Taihua Wu,etc.Blocking core fucosylation of TGF-β1 receptors downregulates their functions and attenuates the epithelial-mesenchymal transition of renal tubular cells.[J].2011,69(4).
- [4]Wen, Xinyu,Liu, Anqi,Yu, Chngqing,etc.Inhibiting post-translational core fucosylation prevents vascular calcification in the model of uremia[J].2016,79.
- [5]Xiangchun, Wang,Jianguo, Gu,Eiji, Miyoshi,etc.Phenotype changes of Fut8 knockout mouse: core fucosylation is crucial for the function of growth factor receptor(s).[J].2006,417.
- [6]Li, Wenzhe,Yu, Rui,Ma, Biao,etc.Core Fucosylation of IgG B Cell Receptor Is Required for Antigen Recognition and Antibody Production[J].2015,194(6).
- [7]Mehrotra, Rajnish,Devuyst, Olivier,Davies, Simon J.,etc.The Current State of Peritoneal Dialysis[J].2016,27(11).
- [8]Kippe, Jordyn M.,Mueller, Toni M.,Haroutunian, Vahram,etc.Abnormal N-acetylglucosaminyltransferase expression in prefrontal cortex in schizophrenia[J].2015,166(1).
- [9]Mueller, Toni M.,Yates, Stefani D.,Haroutunian, Vahram,etc.Altered fucosyltransferase expression in the superior temporal gyrus of elderly patients with schizophrenia[J].2017,182.
- [10]Ueno,T.,Nakashima,A.,Doi,S.,etc.Mesenchymal stem cells ameliorate experimental peritoneal fibrosis by suppressing inflammation and inhibiting TGF-β1 signaling[J].2013,84(2).
- [11]Pranali Patel,Judy West-Mays,Martin Kolb,etc.Platelet derived growth factor B and epithelial mesenchymal transition of peritoneal mesothelial cells[J].2010,29(2).
- [12]Luiz S, Aroeira,Abelardo, Aguilera,José A, Sánchez-Tomero,etc.Epithelial to mesenchymal transition and peritoneal membrane failure in peritoneal dialysis patients: pathologic significance and potential therapeutic interventions.[J].2007,18.
- [13]Wang, Yuqin,Fukuda, Tomohiko,Isaji, Tomoya,etc.Loss of alpha 1,6-fucosyltransferase inhibits chemical-induced hepatocellular carcinoma and tumorigenesis by down-regulating several cell signaling pathways[J].2015,29(8).
- [14]Venkatachalam,M.A.,Weinberg,J.M..New wrinkles in old receptors: Core fucosylation is yet another target to inhibit TGF-β signaling[J].2013,84(1).
- [15]Kobayashi,S.,Maeno,T.,Ueno,M.,etc.Sensitivity of heterozygous α1,6-fucosyltransferase knock-out mice to cigarette smoke-induced emphysema: Implication of aberrant transforming growth factor-β signaling and matrix metalloproteinase gene expression[J].2012,287(20).
- [16]Busnadiego, Oscar,Loureiro-Alvarez, Jesus,Sandoval, Pilar,etc.A Pathogenetic Role for Endothelin-1 in Peritoneal Dialysis-Associated Fibrosis[J].2015,26(1).
- [17]Cordy,P.,Blake,P.,Jain,A.K.,etc.Global trends in rates of peritoneal dialysis[J].2012,23(3).
- [18]Shao, Kang,Chen, Zhong Yi,Gautam, Suraj,etc.Posttranslational modification of E-cadherin by core fucosylation regulates Src activation and induces epithelial-mesenchymal transition-like process in lung cancer cells[J].2016,26(2).
- [19]Devuyst O,Margetts PJ,Topley N.The pathophysiology of the peritoneal membrane.[J].2010,21(7).
- [20]Liappas, Georgios,Tirma Gonzalez-Mateo, Guadalupe,Sanchez-Diaz, Raquel,etc.Immune-Regulatory Molecule CD69 Controls Peritoneal Fibrosis[J].2016,27(12).
- [21]Jesús, Loureiro,Abelardo, Aguilera,Rafael, Selgas,etc.Blocking TGF-β1 protects the peritoneal membrane from dialysate-induced damage.[J].2011,22.
- [22]Takahashi, M,Kuroki, Y,Taniguchi, N,etc.Core fucose and bisecting GlcNAc, the direct modifiers of the N-glycan core: their functions and target proteins[J].2009,344(12).
- [23]Shen,N.,Lin,H.,Wu,T.,etc.Inhibition of TGF-β1-receptor posttranslational core fucosylation attenuates rat renal interstitial fibrosis[J].2013,84(1).
- [24]Wang X,Inoue S,Gu J,etc.From The Cover: Dysregulation of TGF-{beta}1 receptor activation leads to abnormal lung development and emphysema-like phenotype in core fucose-deficient mice.[J].2005,102(44).
- [25]Raffaele, Strippoli,Roberto, Moreno-Vicente,Cecilia, Battistelli,etc.Molecular Mechanisms Underlying Peritoneal EMT and Fibrosis.[J].2016.