Jessup J M, Qi K F, Kanellopoulos K, Cleary K, Hickey C, Reading C L
Department of Surgery, University of Texas M.D. Anderson Hospital and Tumor Institute, Houston 77030.
J Cell Biochem. 1988 Jun;37(2):193-202. doi: 10.1002/jcb.240370206.
Recent interest has focused on fucosylated epitopes expressed on human neoplasms. The plant lectin Ulex europus agglutinin, Type I (UEA) binds fucosylated oligosaccharides, while UEA-reactive substances have a tissue distribution similar to carcinoembryonic antigen (CEA). We sought to determine if UEA reacted with CEA in extracts of fresh primary and metastatic colorectal carcinomas and paired normal tissues. The extracts were electrophoretically transferred to nitrocellulose membranes after the proteins were separated by SDS-PAGE in 10% polyacrylamide gels. The transfer membranes were then stained with peroxidase-conjugated UEA (UEA-P) or antibody to CEA (CEA-P). UEA-P reacted with a 170-190-kDa band in extracts of 22 of 30 primary tumors, 10 of 12 metastases, but only 1 of 5 villous adenomas. UEA-P generally did not react with normal colon or liver extracts. UEA-P also did not bind to 170-190-kDa molecules in Western transfers of a breast carcinoma metastatic to bowel and a focal nodular hyperplasia of liver. CEA-P displayed similar reactivity and detected CEA in a tumor extract negative for UEA. Fucose blocked binding of UEA-P to Western transfers of tumor extracts. CEA-P reacted with a 170-190-kDa substance in tumor extracts eluted with fucose from a column of immobilized UEA. Thus, UEA reacts with fucosylated oligosaccharides on most, but not all, species of CEA and may be a useful adjunct to anti-CEA immunohistochemistry.
最近的研究兴趣集中在人类肿瘤上表达的岩藻糖基化表位。植物凝集素欧洲荆豆凝集素I型(UEA)能结合岩藻糖基化寡糖,而UEA反应性物质的组织分布与癌胚抗原(CEA)相似。我们试图确定UEA是否与新鲜原发性和转移性结直肠癌及配对正常组织提取物中的CEA发生反应。蛋白质在10%聚丙烯酰胺凝胶中通过SDS-PAGE分离后,提取物被电泳转移到硝酸纤维素膜上。然后将转移膜用辣根过氧化物酶偶联的UEA(UEA-P)或CEA抗体(CEA-P)染色。UEA-P在30例原发性肿瘤中的22例提取物、12例转移瘤中的10例提取物中与一条170 - 190 kDa的条带发生反应,但在5例绒毛状腺瘤中只有1例发生反应。UEA-P一般不与正常结肠或肝脏提取物发生反应。UEA-P在转移至肠道的乳腺癌和肝脏局灶性结节性增生的Western印迹中也不与170 - 190 kDa分子结合。CEA-P显示出类似的反应性,并在UEA检测为阴性的肿瘤提取物中检测到CEA。岩藻糖可阻断UEA-P与肿瘤提取物Western印迹的结合。CEA-P与用岩藻糖从固定化UEA柱上洗脱的肿瘤提取物中的170 - 190 kDa物质发生反应。因此,UEA与大多数但并非所有种类的CEA上的岩藻糖基化寡糖发生反应,可能是抗CEA免疫组织化学的有用辅助手段。