基于MALDI-TOF质谱技术分析人宫颈癌HeLa细胞N-糖链结构轮廓  

Analysis of N-Glycans Profiling from HeLa Cells by Matrix-assisted Laser Desorption/Ionization Time-of-flight Mass Spectrometry

作　　者：王培培[1] 于广利[1] 焦广玲[1] 赵峡[1] 李静[1] 柴文刚[1] 

机构地区：[1]中国海洋大学医药学院,海洋药物教育部重点实验室,山东省糖科学与糖工程重点实验室,青岛266003

出　　处：《高等学校化学学报》2012年第6期1233-1238,共6页Chemical Journal of Chinese Universities

基　　金：国家“九七三”计划项目(批准号:2003CB716401);国际科技合作专项(批准号:2007DFA30980);教育部长江学者与创新团队发展计划(批准号:IRT0944);国家自然科学基金(批准号:31070724)资助

摘　　要：以微量HeLa细胞(107个)为对象,经细胞裂解、还原羧甲基化、胰酶降解和Oasis-HLB柱提取分离得到总糖肽后,用PNGase F酶解释放N-糖链.对所得N-糖链用Sep-Pak C18柱纯化后进行完全甲基化衍生,再采用基质辅助激光解吸电离-飞行时间质谱(MALDI-TOF MS)分析HeLa细胞表面N-糖链的结构轮廓.结果表明,在获得的34种N-糖链中,除高甘露糖型、二天线、三天线、四天线和五天线等N-糖链外,还出现了在某种程度上与肿瘤发生转移相关的特殊平分型和Lewis结构.利用MALDI-TOF MS技术可快速分析微量癌细胞表面N-糖链的结构轮廓,为进一步寻找肿瘤糖链标志物及肿瘤的早期预防诊断提供技术支持.The structure of N-glycans on cell surface was abnormal in the process of cancer, which was closely related to the invasion and metastasis of tumors. Total glycopeptides were obtained from trace HeLa cells（ 107） after cell lyses, reduction and acetamidation, trypsin digestion and Oasis-HLB cartridge purification. N-glycans were released by PNGase F digestion and purified by a Sep-Pak C18 cartridge. The structural of N-glycans profiling was first analyzed by positive-ion matrix-assisted laser desorption/ionization time-of-flight mass spectrometry （ MALDI-TOF MS） after permethylation. The 34 types of N-glycans obtained from HeLa cell included both high-mannose and complex （bi-, tri-, tetra- and penta-antennary） types, some of the latter con- taining bisecting and Lewis determinants, which were relevant to tumor metastasis in same way. The MALDI- TOF MS technology was suitable for rapid analysis of N-glycans profiling on cancer cell surface in trace amount, in order to find new glycans-makers and more foundation for prevention and diagnosis in early stage of tumors.

关 键 词：肿瘤 HELA细胞 N-糖链 基质辅助激光解吸电离-飞行时间质谱 

分 类 号：O629.12[理学—有机化学] O657[理学—化学]