摘要
以培养的原发性肝细胞癌Hep G2细胞和正常肝细胞L02为研究对象,用细胞裂解液提取总蛋白,然后采用Carlson还原性β-消除法释放O-糖链,以阳离子交换柱结合C18柱纯化分离O-糖链,用电喷雾电离质谱(ESI-MS)和串联质谱(MS/MS)对O-糖链进行序列鉴定,以β-环糊精为内标对2种细胞系的O-糖链进行定量比较分析.结果表明,在肝癌细胞系Hep G2中检测到10种O-糖链,正常细胞系L02中检测到9种O-糖链,其中9种O-糖链是2种细胞系中共有的,但Hep G2中存在癌细胞中特有的缩短的O-糖链N1A1(Neu Ac-Gal NAc,sialyl Tn抗原).t检验结果表明,Hep G2与L02相比,在检测到的10种O-糖链中有5种的含量具有极显著性差异(P<0.01),2种的含量具有显著性差异(P<0.05).
HepG2（ a primary hepatocellular carcinoma cell line） and L02 （ones derived from normal liver tissue） cells were chosen as model cell lines for research. The O-glycans of the total proteins extracted from HepG2 and 12）2 cells were released by Carlson reduetive β-elimination. The released O-glycans previously / purified by Dowex 50 WX8-400 cation exchange resin and C18 cartridges were identified by electrospray ionization mass spectrometry（ESI-MS） and MS/MS. For eomparision studies,β-cyclodextrin was used as the internal standard for relative quantitative analysis of the O-glycans derived from HepG2 and L02 cells by MS. As results, 10 O-glycans were observed in HepG2 cell line and 90-glycans were detected in L02 cell line. Moreover, 90-glycans were observed in both HepG2 and L02 cells, wherears 1 truncated O-glycan assigned as H1A1 （NeuAc-GalNAc, sialyl Tn antigen, ubiquitous in cancer cells） , was only found in HepG2 cells, t-Test results show that 5 and 20-glycans in HepG2 cells have significant differences （P〈0. 01 and P 〈 0. 05, recpectively）, when compared to those of 1332 cells. Our studies show methodological significance in structural investigation of O-glycans expressed in hepatocellular carcinoma and early biomarker discovery in clinical diagnose.
出处
《高等学校化学学报》
SCIE
EI
CAS
CSCD
北大核心
2015年第7期1275-1281,共7页
Chemical Journal of Chinese Universities
基金
国家自然科学基金(批准号:31370804
31170773)
陕西省教育厅自然科学专项基金(批准号:2013JK0714)资助~~
作者简介
张英，女，博士，讲师，主要从事糖链的荧光标记和质谱分析研究．E-mail：zhangying@nwu．edu．cn
王仲孚，男，博士，教授，博士生导师，主要从事糖生物学与糖工程学研究．E—mail：wangzhf@nwu．edu．cn
