Objective: To investigate the correlation of SphK1, FAK, Musashi 1 and CA199 expression with angiogenesis and epithelial-mesenchymal transition in surgically removed colon cancer lesions. Methods: A total of 60 patien...Objective: To investigate the correlation of SphK1, FAK, Musashi 1 and CA199 expression with angiogenesis and epithelial-mesenchymal transition in surgically removed colon cancer lesions. Methods: A total of 60 patients with colon cancer who underwent radical operation in our hospital between August 2015 and August 2017 were selected, intraoperative colon cancer tissue samples were collected as colon cancer group, and normal tissue specimens adjacent to carcinoma were collected as adjacent tissue group. Fluorescence quantitative PCR was adopted to determine the expression levels of SphK1, FAK, Musashi 1, CA199 as well as the genes related to angiogenesis and epithelial-mesenchymal transition in colon tissues with different properties. Results: SphK1, FAK, Musashi 1 and CA199 mRNA expression in colon cancer group were higher than those in adjacent tissue group;angiogenesis-related genes ANGPTL4, Apelin-13, DLL1, VEGF and HIF-α mRNA expression were higher than those in adjacent tissue group whereas TSP-1 mRNA expression was lower than that in adjacent tissue group;epithelial-mesenchymal transition-related gene E-cadherin mRNA expression was lower than that in adjacent tissue group whereas Vimentin, N-cadherin, Twist and Snail mRNA expression were higher than those in adjacent tissue group. Correlation analysis showed that the SphK1, FAK, Musashi 1 and CA199 expression in colon cancer tissues were directly correlated with the angiogenesis genes and epithelial-mesenchymal transition genes. Conclusion: SphK1, FAK, Musashi 1 and CA199 genes are abnormally expressed in colon cancer tissues and their expression levels are directly correlated with tumor angiogenesis and epithelial-mesenchymal transition process.展开更多
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 protei...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.展开更多
文摘Objective: To investigate the correlation of SphK1, FAK, Musashi 1 and CA199 expression with angiogenesis and epithelial-mesenchymal transition in surgically removed colon cancer lesions. Methods: A total of 60 patients with colon cancer who underwent radical operation in our hospital between August 2015 and August 2017 were selected, intraoperative colon cancer tissue samples were collected as colon cancer group, and normal tissue specimens adjacent to carcinoma were collected as adjacent tissue group. Fluorescence quantitative PCR was adopted to determine the expression levels of SphK1, FAK, Musashi 1, CA199 as well as the genes related to angiogenesis and epithelial-mesenchymal transition in colon tissues with different properties. Results: SphK1, FAK, Musashi 1 and CA199 mRNA expression in colon cancer group were higher than those in adjacent tissue group;angiogenesis-related genes ANGPTL4, Apelin-13, DLL1, VEGF and HIF-α mRNA expression were higher than those in adjacent tissue group whereas TSP-1 mRNA expression was lower than that in adjacent tissue group;epithelial-mesenchymal transition-related gene E-cadherin mRNA expression was lower than that in adjacent tissue group whereas Vimentin, N-cadherin, Twist and Snail mRNA expression were higher than those in adjacent tissue group. Correlation analysis showed that the SphK1, FAK, Musashi 1 and CA199 expression in colon cancer tissues were directly correlated with the angiogenesis genes and epithelial-mesenchymal transition genes. Conclusion: SphK1, FAK, Musashi 1 and CA199 genes are abnormally expressed in colon cancer tissues and their expression levels are directly correlated with tumor angiogenesis and epithelial-mesenchymal transition process.
基金This work was supported by a grant from the National Natural Science Foundation of China （No. 81530021）.
文摘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.
