凝集素点印迹法分析胆汁糖蛋白糖链结构诊断胆管癌

The Diagnosis of Bile Duct Carcinoma by Analysis of Sugar Chain of Glycoprotein in Bile with Lectin Dot blotting

目的 研究胆管癌胆汁中的肿瘤相关糖蛋白N 连接型糖链结构及其诊断价值。方法 将 31例胆管癌胆汁和 13例良性胆道疾病胆汁直接点在硝酸纤维膜 (NC)上 ,采用辣根过氧化物酶 (HRP)标记的刀豆凝集素 (ConA )、欧曼陀罗凝集素 (DSA )、麦胚凝集素 (WGA )、小扁豆凝集素 (LCA )进行点印迹分析。结果 胆管癌组和良性组ConA、DSA、LCA和WGA阳性率 (阳性例数 /总例数 )分别为 80 .6 % (2 5 /31)和 76 .9% (10 /13)、74.2 % (2 3/31)和 0 (0 /13)、70 .0 % (2 2 /31)和 15 .4% (2 /13)、80 .6 % (2 5 /31)和 2 3 .1% (3/13) ,胆管癌组DSA、LCA和WGA阳性率高于良性组 (P <0 .0 5 )。结论 胆管癌胆汁糖蛋白复杂型糖链增多 ,其核心岩藻糖和唾液酸含量丰富 ,可能出现偏二天线以及天线数的增加。

Objective To study the N linked oligosaccharides of tumor related glycoprotein in bile and its diagnostic value for bile duct carcinoma.Methods The bile samples from 31 cases of cholangiocarcinomas and 13 cases of benign biliary diseases were dropped onto nitrocellulose membrane respectively,followed by lectin dot blotting with horseradish peroxidase labeled lectins,including ConA(coneanavalia ensifomis agglutinin),DSA(datura stramonium agglutinin),WGA(triticum valgaris agglutinin) and LCA(lens culinaris agglutinin).Results The positive rates of ConA,DSA,LCA and WGA in cholangiocarcinoma group and benign group were 80.6%(25/31) vs 76.9%(10/13),74.2%(23/31) vs 0(0/13),70.0%(22/31) vs 15.4%(2/13),80.6%(25/31) vs 23.1% (3/13),respectively.The positive rates of DSA,LCA and WGA in malignant group were higher than in benign one( P < 0.05 ).Conclusion Complex type asparagine (Asn) linked oligosaccharides are found in bile glycoprotein of bile duct carcinoma,with notably increased sialylation and core fucose,maybe aberrant bi antennary N glycan and increasement of the amount of antennary.Lectin dot blotting of bile glycoprotein may be an effective method for the diagnosis of cholangiocarcinoma.