Dwek M V, Ross H A, Streets A J, Brooks S A, Adam E, Titcomb A, Woodside J V, Schumacher U, Leathem A J
Department of Surgery, Royal Free and University College London Medical School, London, United Kingdom.
Int J Cancer. 2001 Mar 20;95(2):79-85. doi: 10.1002/1097-0215(20010320)95:2<79::aid-ijc1014>3.0.co;2-e.
Predicting long-term outcome after breast-cancer diagnosis remains problematic, particularly for patients with clinically small, axillary lymph node- negative tumours. Evidence suggests that the lectin Helix pomatia agglutinin (HPA) identifies oligosaccharides associated with poor-prognosis cancer. Our aim was to identify oligosaccharides that bind HPA in aggressive breast cancers. Breast-cancer cell lines (MCF-7, BT-549 and BT-20) and a cell line from human milk (HBL-100), which showed a range of HPA-binding intensities, were used to extract HPA-binding glycoproteins. Oligosaccharides were released using anhydrous hydrazine and separated on a range of HPLC matrices. We investigated whether HPA-binding oligosaccharides from cell lines were present in human breast-cancer tissues, using 69 breast-cancer specimens from patients with between 5 and 10 years' follow-up. A monosialylated oligosaccharide was over-expressed in the cell line that bound HPA strongly. Further analysis by normal-phase HPLC showed that the 2-aminobenzamide-conjugated oligosaccharide had a hydrodynamic volume of 4.58 glucose units (HPAgly1). Increased expression of HPAgly1 was associated with HPA staining of breast-cancer specimens (Student's t-test p = 0.025). Analysis of oligosaccharide levels and disease-free survival after treatment for breast cancer indicated a shorter disease-free interval for patients with elevated levels of HPAgly1. This is the first time that histochemical lectin staining has been correlated with biochemical mapping of oligosaccharides. Using this approach, we have identified a monosialylated HPA lectin-binding oligosaccharide present in breast-cancer cells grown in vitro which is elevated in breast-cancer specimens that bind the lectin.
预测乳腺癌诊断后的长期预后仍然存在问题，尤其是对于临床分期为小肿瘤、腋窝淋巴结阴性的患者。有证据表明，凝集素苹果蜗牛凝集素（HPA）可识别与预后不良的癌症相关的寡糖。我们的目的是鉴定侵袭性乳腺癌中与HPA结合的寡糖。乳腺癌细胞系（MCF-7、BT-549和BT-20）以及人乳腺细胞系（HBL-100），它们显示出一系列HPA结合强度，用于提取与HPA结合的糖蛋白。使用无水肼释放寡糖，并在一系列HPLC基质上进行分离。我们使用来自随访5至10年患者的69份乳腺癌标本，研究了细胞系中与HPA结合的寡糖是否存在于人类乳腺癌组织中。一种单唾液酸化寡糖在与HPA强烈结合的细胞系中过度表达。通过正相HPLC进一步分析表明，2-氨基苯甲酰胺缀合的寡糖的流体力学体积为4.58个葡萄糖单位（HPAgly1）。HPAgly1表达增加与乳腺癌标本的HPA染色相关（Student's t检验p = 0.025）。分析乳腺癌治疗后寡糖水平和无病生存期表明，HPAgly1水平升高的患者无病间隔较短。这是首次将组织化学凝集素染色与寡糖的生化图谱相关联。使用这种方法，我们鉴定出一种存在于体外培养的乳腺癌细胞中的单唾液酸化HPA凝集素结合寡糖，其在与凝集素结合的乳腺癌标本中升高。
