乳腺癌组织N-连接糖链的分析

Analysis of N-linked glycans in breast carcinoma

目的： 解析乳腺癌组织的特异性糖链结构，为乳腺癌的早期诊断提供可能标志物。方法：收集2014年5月至2014年9月在陕西省肿瘤医院乳腺科手术切除的乳腺癌患者组织标本15例及距癌组织边缘3-5 cm的癌旁组织标本5例，另选取乳腺小叶增生组织标本4例，提取组织蛋白质，利用滤膜辅助的糖蛋白糖链释放方法获得N-连接糖链，应用基质辅助激光解析电离飞行时间质谱（MALDITOF/TOFMS）技术分析乳腺癌组织、癌旁组织和正常乳腺组织中糖蛋白N连接糖链类型分布和含量变化情况。结果：乳腺癌组织、癌旁组织和正常乳腺组织中分别鉴定到38、34和22个糖链。与癌旁组织及正常乳腺组织相比，癌症组织中复杂型、高甘露糖型、平分型、岩藻糖化和二天线型糖链的含量除高甘露糖型明显升高外，其他均显著降低（P〈0.05或P〈0.01）。乳腺癌组织中相对分子质量分别为1 580.455和1 742.490的2种高甘露糖型N糖链的含量显著高于正常乳腺组织（P〈0.05或P〈0.01）、相对分子质量分别为1 647.473和1 850.583的2种复杂型N-糖链的含量则显著低于正常乳腺组织（P〈0.05或P〈0.01）。这4种糖链经ROC曲线分析显示具有较高检测准确性（P〈0.05或P〈0.01）。结论：发现的4种乳腺癌组织特异性的N糖链有成为早期诊断和靶向治疗标志物的潜力。

Objective：To analyze structures of specific glycans in breast carcinoma, in order to provide potential early diagnostic markers for breast carcinoma. Methods： Fifteen specimens of breast carcinoma and five specimens of tissues adjacent to the carcinoma margin 3-5 cm surgical resected from the patients with breast cancer who were hospitalized in Department of Galactophore, Tumor Hospital of Shaanxi Province during May 2014 to September 2014 were collected. In addition, four specimens of breast lobular hyperplasia tissues were selected. Tissue proteins were extracted. Using filter membrane assisted method for the release of glycoprotein glycans, N-linked glycans were obtained. Type distributions and content changes of glycoprotein N-linked glycans in breast carcinoma, adjacent breast carcinoma and normal breast tissues were analyzed by matrix assisted laser desorption ionization time of flight mass spectrometry （MALDI-TOF/TOF-MS） tech- nique. Results： Thirtyeight, thirty four and twentytwo N-linked glycans were respectively identified in the breast cancer tissues, the adjacent tissues, and the normal breast tissues. Comparing with the paracarcinoma tissues and the normal breast tissues, contents of complex, high mannose, bisecting, fucosylated and two attenna types of N-linked glycans in the breast carcinoma tissues significantly decreased except for that content of high mannose type N-linked glycans obviously in- creased （ P 〈 0.05 or P 〈 0.01 ）. In the breast carcinoma tissue, contents of two types of high mannose type N-linked gly- cans, relative molecular masses of that were 1 580. 455 and 1 742. 490 respectively, were significantly higher than those in the normal breast tissues （ P 〈 0.05 or P 〈 0.01 ）, and contents of two types of complex type N-linked glycans, relative molecular masses of that were 1 647. 473 and 1 850. 583 respectively, were obviously lower than those in the normal breasttissues （P 〈 0.05 or P 〈 0.01 ）. ROC curve analysis showed that the 4 kinds of N-linked glycanshave high detection accuracy （P 〈0.05 or P 〈 0.01 ）. Conclusion： Four types of specific N-linked glycans found in the breast carcinoma could be expected to become potential markers for early diagnosis and target therapy of the breast carcinoma.