N-glycosylation of colorectal cancer tissues: a liquid chromatography and mass spectrometry-based investigation.
N-glycosylation of colorectal cancer tissues: a liquid chromatography and mass spectrometry-based investigation.
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DOI：
10.1074/mcp.m111.011601
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发表时间：
2012-09
期刊：
影响因子：
--
通讯作者：
Wuhrer M
中科院分区：
文献类型：
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作者：
Balog CI;Stavenhagen K;Fung WL;Koeleman CA;McDonnell LA;Verhoeven A;Mesker WE;Tollenaar RA;Deelder AM;Wuhrer M
结直肠癌是全球第三大常见癌症，每年∼发病率为100万例，年死亡率为∼655,000人。迫切需要确定新的靶点，以开发更灵敏、可靠和特异的结肠癌早期检测方法。翻译后修饰在肿瘤进展和肿瘤免疫监视中发挥着重要作用。在本研究中，我们比较了13例结直肠癌肿瘤组织和相应的对照结肠组织的N-糖链图谱。用酶法释放、纯化N-葡聚糖，并用2-氨基苯甲酸标记。采用HILIC-HPLC型荧光检测联用和MALDI-TOF-MS负模式联用对等分进行了分析。用偏最小二乘判别分析研究结直肠癌N-糖基化的变化，对HILIC-HPLC和MALDI-TOF-MS数据都有很好的分离和预测能力。在结构鉴定方面，结合了正模式ESI-离子陷阱-MS/MS和负模式MALDI-TOF/TOF-MS的信息。在具有较高分离力的特征中，含有二等分GlcNAc的结构在肿瘤组织中减少，而在肿瘤组织中含有唾液酸化Lewis表位的硫酸多糖、低聚糖苷和含有唾液酸化Lewis表位的多糖增加。此外，在肿瘤标本中还检测到核心岩藻糖基化的高甘露糖N-糖链。综上所述，HILIC和MALDI-TOF-MS对N-糖链的分析与多元统计分析相结合，显示了其在结直肠癌组织中识别N-糖基化变化的潜力，并提供了可能被用作候选生物标记物的新线索。
Colorectal cancer is the third most common cancer worldwide with an annual incidence of ∼1 million cases and an annual mortality rate of ∼655,000 individuals. There is an urgent need for identifying novel targets to develop more sensitive, reliable, and specific tests for early stage detection of colon cancer. Post-translational modifications are known to play an important role in cancer progression and immune surveillance of tumors. In the present study, we compared the N-glycan profiles from 13 colorectal cancer tumor tissues and corresponding control colon tissues. The N-glycans were enzymatically released, purified, and labeled with 2-aminobenzoic acid. Aliquots were profiled by hydrophilic interaction liquid chromatography (HILIC-HPLC) with fluorescence detection and by negative mode MALDI-TOF-MS. Using partial least squares discriminant analysis to investigate the N-glycosylation changes in colorectal cancer, an excellent separation and prediction ability were observed for both HILIC-HPLC and MALDI-TOF-MS data. For structure elucidation, information from positive mode ESI-ion trap-MS/MS and negative mode MALDI-TOF/TOF-MS was combined. Among the features with a high separation power, structures containing a bisecting GlcNAc were found to be decreased in the tumor, whereas sulfated glycans, paucimannosidic glycans, and glycans containing a sialylated Lewis type epitope were shown to be increased in tumor tissues. In addition, core-fucosylated high mannose N-glycans were detected in tumor samples. In conclusion, the combination of HILIC and MALDI-TOF-MS profiling of N-glycans with multivariate statistical analysis demonstrated its potential for identifying N-glycosylation changes in colorectal cancer tissues and provided new leads that might be used as candidate biomarkers.