O-GlcNAc糖基化修饰对胃癌细胞黏附能力的影响  

Effects of O-GlcNAcylation on the adhesion ability of gastric cancer cells

作　　者：赵其杰 周时胜 娄文辉 钱晖[1] 徐志伟 ZHAO Qijie;ZHOU Shisheng;LOU Wenhui;QIAN Hui;XU Zhiwei(School of Medicine,Jiangsu University,Zhenjiang Jiangsu 212013,China)

机构地区：[1]江苏大学医学院,江苏镇江212013

出　　处：《江苏大学学报(医学版)》2025年第2期102-109,117,共9页Journal of Jiangsu University:Medicine Edition

基　　金：国家自然科学基金资助项目(32000903);江苏大学高级人才启动基金资助项目(20JDG48)。

摘　　要：目的:探讨O连接的N-乙酰葡萄糖胺(O-linked N-acetylglucosamine,O-GlcNAc)糖基化修饰对胃癌细胞和细胞外基质之间黏附能力的影响以及潜在的作用机制。方法:通过UALCAN数据库分析人泛癌和胃癌组织中己糖胺生物合成途径(hexosamine biosynthesis pathway,HBP)中谷氨酰胺-6-磷酸果糖转氨酶1(glutamine-fructose-6-phosphate transaminase 1,GFPT1)、氨基葡萄糖磷酸N-乙酰转移酶1(glucosamine-phosphate N-acetyltransferase 1,GNPNAT1)、磷酸葡萄糖变位酶3(phosphoglucomutase 3,PGM3)、UDP-N-乙酰葡糖胺焦磷酸化酶1(UDP-N-acetylglucosamine pyrophosphorylase 1,UAP1)、N-乙酰氨基葡萄糖转移酶(N-acetylglucosamine transferase,OGT)和O-GlcNAc水解酶(O-GlcNAcase,OGA)mRNA相对表达量;通过DepMap数据库分析胃癌HGC-27、AGS和SNU-1细胞中GFPT1、GNPNAT1、PGM3、UAP1、OGT和OGA mRNA相对表达量;通过蛋白免疫印迹检测人胃黏膜上皮GES-1细胞和胃癌HGC-27、AGS和SNU-1细胞中GFPT1、OGT和OGA蛋白相对表达水平以及O-GlcNAc糖基化修饰水平;通过基因富集分析检测OGT和OGA差异表达的信号通路;通过黏附实验检测人胃黏膜上皮GES-1细胞和胃癌HGC-27、AGS和SNU-1细胞的黏附能力。通过OGT抑制剂OSMI-1和OGA抑制剂Thiamet G(TMG)分别降低和升高胃癌HGC-27和SNU-1细胞O-GlcNAc糖基化修饰水平,随后采用黏附实验检测其黏附能力。结果:GFPT1、GNPNAT1、PGM3、OGT和OGA mRNA表达量在泛癌尤其是胃癌组织中较癌旁组织明显升高(P<0.05)。与胃癌HGC-27和SNU-1细胞相比,胃癌AGS细胞中GFPT1、GNPNAT1、PGM3和UAP1 mRNA表达量较高,但是OGT和OGA mRNA表达量较低。GFPT1蛋白表达量在人胃黏膜上皮细胞和不同胃癌细胞系中无明显区别,但是OGT和OGA蛋白表达量在胃癌HGC-27和SNU-1细胞中相较于人胃黏膜上皮GES-1细胞明显升高(P<0.01)。O-GlcNAc相对表达水平在胃癌SNU-1细胞中的表达量明显高于人胃黏膜上皮GES-1细胞和胃癌HGC-27和AGS细胞(P<0.01)。基�Objective:To investigate the effect of O-linked N-acetylglucosamine(O-GlcNAc)glycosylation on the adhesion between gastric cancer cells and extracellular matrix and its potential mechanisms.Methods:The mRNA expressions of glutamine-fructose-6-phosphate transaminase 1(GFPT1),glucosamine-phosphate N-acetyltransferase 1(GNPNAT1),phosphoglucomutase 3(PGM3),UDP-N-acetylglucosamine pyrophosphorylase 1(UAP1),N-acetylglucosamine transferase(OGT)and O-GlcNAcase(OGA)in pan-cancer and human gastric cancer tissues were analyzed by UALCAN database.The relative mRNA expression levels of GFPT1,GNPNAT1,PGM3,UAP1,OGT and OGA in gastric cancer HGC-27,AGS,and SNU-1 cells were analyzed by DepMap database.The relative expression levels of GFPT1,OGT and OGA proteins and the glycosylation modification levels of O-GlcNAc in human gastric mucosal epithelial GES-1 cells and gastric cancer HGC-27,AGS and SNU-1 cells were detected by Western blotting.The signaling pathways enriched by the gastric cancer samples with differential expression of OGT and OGA were explored by Gene Set Enrichment Analysis(GSEA).The adhesion ability of human gastric mucosal epithelial GES-1 cells and gastric cancer HGC-27,AGS,and SNU-1 cells was detected by cell adhesion assay.OGT inhibitor OSMI-1 and OGA inhibitor Thiamet G(TMG)were used to decrease or increase O-GlcNAc glycosylation modification levels of gastric cancer cells HGC-27 and SNU-1 cells,respectively,and their adhesion ability was detected by adhesion assay.Results:The mRNA expression levels of GFPT1,GNPNAT1,PGM3,OGT,and OGA were significantly elevated in pan-cancer tissues,especially gastric cancer,compared with those in adjacent tissues(P<0.05).Compared with HGC-27 and SNU-1 cells,the mRNA expressions of GFPT1,GNPNAT1,PGM3 and UAP1 in AGS cells of gastric cancer were higher,but the mRNA expressions of OGT and OGA were lower.The expression of GFPT1 protein was not significantly different among human gastric mucosal epithelial cells and three gastric cancer cell lines,but the expression of OGT and OGA

关 键 词：O连接的N-乙酰葡萄糖胺(O-GlcNAc) 胃癌 细胞黏附 己糖胺生物合成途径 细胞外基质 

分 类 号：R735.2[医药卫生—肿瘤]