摘要
目的应用凝集素芯片技术寻找肝癌细胞表面转移相关的特征性糖谱。方法应用肝细胞生长因子(hepatocyte growth factor,HGF)诱导建立肝癌上皮间质转化(epithelial mesenchymal transition,EMT)细胞模型。通过凝集素芯片比较诱导前后细胞膜的糖谱改变,采用凝集素印迹和荧光细胞凝集素免疫组化方法验证芯片结果。结果诱导后细胞对凝集素ACL、BPL、JAC、MPL、PHA-E、SBA和SNA的亲和作用减弱,而对凝集素AAL、ConA、DBA、GSLⅡ、ECL、HAL、LCA、LTL、NML、NPL、PHA-L、PTLⅡ和WFL的亲和作用增强。提示诱导后肝癌细胞表面出现了黏蛋白T/Tn抗原、平分型N-乙酰葡萄糖胺、α2,6唾液酸和末端α或β连接的N-乙酰半乳糖胺结构减少;而末端和核心岩藻糖、高甘露糖、N-乙酰葡萄糖胺β1,6分支和复杂型寡糖分支结构增多。结论肝癌细胞发生EMT过程中细胞膜表面糖链结构改变,提示糖链结构与肝癌的转移密切相关,为有效控制肝癌转移、改善预后提供了新思路。
Objective To search the specific cell surface glycan profiling related with hepatocellular carcinoma （HCC） metastasis by lectin microarray.Methods Hepatocyte growth factor （HGF） induced epithelial mesenchymal transition （EMT） model in Huh7 cells was established.Cell surface glycan profiling of Huh7 cells treated with or without HGF were compared by lectin microarray,then the results from lectin microarray were verified by lectin blot and fluorescence cell lectinimmunochemistry.Results The binding affinities to 7 lectins ACL,BPL,JAC,MPL,PHAE,SBA and SNA were reduced in HGF induced EMT cells,meanwhile the affinities to bind 13 lectins,AAL,ConA,DBA,GSL Ⅱ,ECL,HAL,LCA,LTL,NML,NPL,PHAL,PTL Ⅱ and WFL,were increased with statistic significance.It implied that the decreased T/Tn-antigen,NA2 and bisecting GlcNAc,Sia α 26 Gal/GalNAc,terminal α or β GalNAc structures,and the increased terminal and core fucose,high mannose,β 1,6 GlcNAc branching and tetraantennary complex oligosaccharides structures may be involved in the changed glycan profiles as EMT process happened in Huh7 cells.Conclusions Cell surface glycan altered in the EMT process and it may be closely related to the metastasis of HCC.This finding provides a new idea for HCC metastasis and prognosis.
出处
《复旦学报（医学版）》
CAS
CSCD
北大核心
2014年第2期198-204,共7页
Fudan University Journal of Medical Sciences
基金
国家重点基础研究发展计划项目(973计划)(2011CB 910604
2013CB 910501)
国家自然科学基金(81060199)
广西自然科学基金(2012GXNSFAA053088)~~
