肝癌标志物——人肝癌组织中碱性磷酸酶糖链结构分析

Analysis of N liked sugar chain structures of ALP from human liver cancer tissue.

裸鼠人肝癌组织（ｈｕｍａｎＬｉｖｅｒｃｅｌｃａｎｃｅｒＴｉｓｓｕｅｉｎＮｕｄｅＭｉｃｅ，ＬＴＮＭ）中碱性磷酸酶（ａｌｋａｌｉｎｅｐｈｏｓ－ｐｈａｔａｓｅ，ＡＬＰ）在ＤＥ５２纤维素离子交换层析柱上均出现两个活性峰，分别由０．１５ｍｏｌ／ＬＮａＣｌ（峰１）和０．５ｍｏｌ／ＬＮａＣｌ（峰２）所洗脱，有些ＬＴＮＭ还会出现１ｍｏｌ／ＬＮａＣｌ洗脱峰。用麦胚凝集素（ｗｈｅａｔｇｅｒｍａｇｇｌｕｔｉｎｉｎ，ＷＧＡ）、小扁豆凝集素（ｌｅｎｓｃｕｌｉｎａｒｉｓａｇｇｌｕｔｉｎｉｎ，ＬＣＡ）、伴刀豆蛋白Ａ（ｃｏｎｃａｎａｖａｌｉｎＡ，ＣｏｎＡ）和蔓陀萝凝集素（ｄａｔｕｒａｓｔｒａ－ｍｏｎｉｕｍａｇｇｌｕｔｉｎｉｎ，ＤＳＡ）固相凝集素亲和层析研究峰１和峰２中ＡＬＰ的糖链结构发现：峰１中ＡＬＰ主要为二天线复杂型糖链，高甘露糖型及杂合型糖链约占３０％，２，４，２－三天线或２，４－偏二天线糖链约占２６％，含核心岩藻糖（Ｆｕｃ）和平分型ＧｌｃＮＡｃ残基的糖链含量分别为５５％和９８％。峰２中ＡＬＰ几乎均为多天线复杂型糖链（９５％），二天线复杂型糖链和高甘露糖型及杂合型糖链含量很少。此峰的ＡＬＰ糖链均不含核心Ｆｕｃ，但含平分型ＧｌｃＮＡｃ残?

To verify if the unique serum ALP with multiple antennary complex type sugar chain comes from liver cancer tissue, WGA(Wheat germ agglutinin)、LCA(Lens culinaris agglutinin)、ConA(concanavalin A) and DSA(Datura stromonium agglutinin) were used to study the alternation of N glycan structures in ALP from human liver cell cancer tissue in nude mice(LTNM). It was found that ALP from LTNM were separated by DEAE Cellulose (DE 52 ) chromatography into two or three active peaks. The first bound peak (76 16±10 86%) was eluted by 0 15 mol/L NaCl and the second bound peak (23 84±11 16%) with 0 5 mol/L NaCl. The third active peak eluted by 1 mol/L NaCl was present in some of LTNM and was too less to study. The first and the second peaks were subjected to WGA、LCA、ConA and DSA immobilized lectin affinity chromatography to analyze their oligosaccharide structures respectively. The results indicated that half of the ALP in the first peak has biantennary complex type sugar chain;30% was high mannose (HM) type and hybrid (Hyb) type.The content of 2,4,2 triantennary and (or) 2,4 biantennary sugar chain (weak bound on DSA) was 26%.55% of the ALP in this peak contained core fucose (Fuc) and 98% contained “Bisecting” GlcNAc residue. The ALP in the second peak contained almost only multiple antennary complex type sugar chain. The content of biantennary complex type sugar chain, HM and Hyb were very low. There were some ALP with core Fuc in this peak. 28% of the ALP in this peak contained “Bisecting” GlcNAc residue. So the conclusion is that the ALP from LTNM does contain multiple antennary complex type sugar chain, and also core Fuc and “Bisecting” GlcNAc residues.