Blocking Posttranslational Core Fucosylation Ameliorates Rat Peritoneal Mesothelial Cell Epithelial-Mesenchymal Transition

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-β receptors and Smad2/3; however, it significantly suppressed the phosphorylation 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.

Inhibiting core fucosylation attenuates glucose-induced peritoneal fibrosis in rats

“…Our recent study has shown that blocking core fucosylation could ameliorate the rat peritoneal mesothelial cell epithelialmesenchymal transition induced by a high glucose solution in vitro. 30 Thus, in this study, we further detected the expression of core fucosylation in the peritoneal membrane of rats with peritoneal fibrosis. We then investigated the effects of inhibiting core fucosylation on the development and progression of peritoneal fibrosis, and finally compared the protective effects of core fucosylation inhibition with imatinib (an available selective PDGF receptor inhibitor).…”

Glycobiology of the Epithelial to Mesenchymal Transition

“…In the latter, FUT8 activated the β-catenin/LEF-1 transcription factor [43], while it was inhibited by miR-198-5p overexpression, resulting in EMT inhibition [44]. FUT8 promoted EMT through core fucosylation of TGF-β receptor in both breast cancer cells [45] and in rat peritoneal mesothelial cells [46]. In the former, FUT8 could be activated by the opioid analgesic fentanyl and promoted EMT and malignancy through the β-catenin pathway [47].…”

Reprogramming of Mesothelial-Mesenchymal Transition in Chronic Peritoneal Diseases by Estrogen Receptor Modulation and TGF-β1 Inhibition

“…Galunisertib monotherapy effectively inhibited tumor growth in a number of human cancer xenografts, and was synergistic with 5-FU and paclitaxel in in vitro gastric cancer cell studies, and with sorafenib in HCC cell lines [141][142][143][144][145]. The inhibition of core fucosylation of Activin receptor-like kinase-5 (ALK-5), TBRII and PDGF receptor by Fut8 small interfering RNA (siRNA) successfully suppresses the activation of TGF-β1/ALK-5/SMAD2/3 and PDGF/ERK signaling pathways and attenuates rat PMC EMT in vitro [146]. This has important implications for the control of GDP/AGE induced CAPD EPS, but also glycosylation related renal interstitial fibrosis and carcinogenesis [132,[147][148][149].…”