高等学校化学学报 ›› 2018, Vol. 39 ›› Issue (9): 1904-1912.doi: 10.7503/cjcu20180143

• 分析化学 • 上一篇    下一篇

基于寡糖代谢工程的正常和肿瘤细胞中Mucin型O-糖链的质谱定性定量比较分析

杨明明, 南丽婧, 晋万军, 王承健, 黄琳娟, 张英(), 王仲孚()   

  1. 西北大学生命科学学院, 西部资源生物与现代生物技术教育部重点实验室, 西安 710069
  • 收稿日期:2018-02-21 出版日期:2018-07-10 发布日期:2018-07-10
  • 作者简介:

    联系人简介: 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方面的研究. Email: wangzhf@nwu.edu.cn; 张 英, 女, 博士, 副教授, 主要从事糖链的荧光标记和质谱分析方面的研究. Email: zhangying@nwu.edu.cn

  • 基金资助:
    国家自然科学基金(批准号: 31670808)、 陕西省自然科学基础研究计划(批准号: 2016JQ3018)和陕西省重点实验室科研项目(批准号: 16JS109, 14JS101)资助.

Quali-quantitative Profiling of Mucin-type O-Glycans in Normal and Tumor Cell Lines Based on Oligosaccharide Metabolic Engineering Combined with Mass Spectrometry

YANG Mingming, NAN Lijing, JIN Wanjun, WANG Chengjian, HUANG Linjuan, ZHANG Ying*(), WANG Zhongfu*()   

  1. Key Laboratory for Western China Resource Biology and Biotechnology, Ministry of Education, College of Life Science, Northwest University, Xi’an 710069, China
  • Received:2018-02-21 Online:2018-07-10 Published:2018-07-10
  • Contact: ZHANG Ying,WANG Zhongfu E-mail:zhangying@nwu.edu.cn;wangzhf@nwu.edu.cn
  • Supported by:
    † Supported by the National Natural Science Foundation of China(No.31670808), the Natural Science Basic Research Program of Shaanxi Province, China(No.2016JQ3018) and the Scientific Research Program Fund for Shannxi Provincial Key Laboratory, China(Nos.16JS109, 14JS101).

摘要:

基于寡糖代谢工程结合质谱技术(MS结合MS/MS), 对4种肿瘤细胞系和1种正常细胞系中的O-糖链进行了定性和相对定量比较分析. 结果表明, 4种肿瘤细胞系HeLa, SMMC-7721, HepG2和MCF-7中分别检测到19, 11, 6和5条O-糖链; 在正常肝细胞系L02中检测到10条O-糖链. 在对肿瘤和正常细胞系中表达的O-糖链进行定性和相对定量比较中发现, 结构组成为N1, H1N1A1和H1N1A2的3种糖链在5种细胞系中均有表达; 肿瘤细胞系表达的O-糖链的种类比正常细胞多, 且岩藻糖基化和唾液酸化程度均高于正常细胞组. 肿瘤细胞系中特有的O-糖链主要有岩藻糖化和唾液酸化修饰的Mucin型Core2结构糖链. MS/MS分析表明, 其中岩藻糖基化修饰的O-糖链结构组成为H3N3F1A2, H4N4F1A2和H5N5F1A1, 唾液酸化修饰的O-糖链结构组成为H5N4A1, H4N4A2和H5N5A2.

关键词: Mucin型O-糖链, 肿瘤细胞, 正常细胞, 寡糖代谢工程, 质谱

Abstract:

The O-glycans on glycoproteins have important biological functions, and their abnormal expressions are closely related to the occurrence and development of several diseases such as tumor. Thus, it is of importance to conduct quali-quantitative profiling of O-glycans expression in normal and disease cells. However, due to the structural complexities and inhomogeneous expression of O-glycans, quali-quantitative analyses of these glycoconjugates still encounter challenges. In this study, based on the oligosaccharide metabolic engineering combined with mass spectrometry, we have profiled and analyzed O-glycans expressed in four tumor cell lines(HeLa, SMMC-7721, HepG2 and MCF-7) and one normal cell lines(L02) quali-quantitatively. The results showed that 19, 11, 6 and 5 O-glycans were observed in four cancer cell lines, HeLa, SMMC-7721, HepG2 and MCF-7 respectively. And 10 O-glycans were detected in one normal cell line(L02). Qualitative and relative quantitative comparison of O-glycans expressed in tumor and normal cell lines showed that three O-glycans(N1, H1N1A1 and H1N1A2) were observed in all cell lines. The number of O-glycans expressed in cancer cell lines is more than those in normal cells. Moreover, these O-glycans were highly fucosylated and sialylated. Further MS/MS data showed that these O-glycans were belonged to Core2 type Mucin O-glycans and their sugar compositions were H3N3F1A2, H4N4F1A2 and H5N5F1A1(fucosylated O-glycans) and H5N4A1, H4N4A2 and H5N5A2(sialylated O-glycans).

Key words: Mucin-type O-glycan, Tumor cell, Normal cell, Oligosaccharides metabolic engineering, Mass Spectrometry

中图分类号: