非转移性和转移性人肝癌血清糖蛋白凝集素亲和谱的比较研究  

Comparative serum glycoproteomics of non-metastatic and metastatic hepatocellular carcinoma by lectin affinity technology

作　　者：陈巧佩[1] 孙淳[2,3] 张舒[2] 康晓楠[3] 李岩[2] 郭坤[2] 孙璐[3] 秦雪[1] 刘银坤[2,3] 

机构地区：[1]广西医科大学第一附属医院临床医学实验部,南宁530021 [2]复旦大学附属中山医院肝癌研究所 [3]复旦大学生物医学研究院,上海200032

出　　处：《复旦学报（医学版）》2011年第5期389-395,共7页Fudan University Journal of Medical Sciences

基　　金：国家重大专项基金项目(2008ZX10002-021);国家重点基础研究发展计划(973计划)项目(2011CB910600);广西自然科学基金项目(2010GXNSFA013149)

摘　　要：目的应用凝集素亲和技术寻找肝癌转移相关特征性血清糖蛋白凝集素亲和谱,并探讨其生物学意义。方法本研究利用含有13种肿瘤相关的凝集素集合芯片技术,比较研究了非转移性和转移性人肝癌血清糖蛋白凝集素亲和谱,同时采用多重凝集素印迹方法验证凝集素芯片结果。计量资料数据采用双样本异方差t检验,进行两两比较。结果研究证明,与非转移人肝癌血清相比,转移性肝癌血清糖蛋白对麦胚凝集素(wheatgerm agglutinin,WGA)、黑接骨木凝集素(sambucus nigra lection,SNA)、红腰豆凝集素(phaseolus vulgariserythroagglutinin,PHAE)和欧曼陀罗凝集素(datura stramonium lectin,DSA)的亲和作用增强(P<0.05)。提示转移性肝癌血清糖蛋白出现了增多的唾液酸(sialic acid,sia)、N-乙酰葡萄糖胺(N-acetyl glucosamine,GlcNAc)、平分型GlcNAc及多天线、偏二天线糖链结构。结论血清糖蛋白聚糖链的变化与肿瘤侵袭和转移密切相关,为临床筛选肝癌侵袭及转移相关的糖标记提供了重要参考。Objective Lectin affinity technology is used to identify specific serum protein glycan profiles of metastatic hepatocellular carcinoma（HCC） compared to non-metastatic HCC,and to explore its biological significance.Methods The high abundance proteins depleted serum proteins were collected and then labeled with fluorescence dye Cy5.The tumor specific lectin microarray containing 13 kinds of tumor-related lectins were employed in this assay.Lectin affinitied glycan profiles of serum protein between non-metastatic and metastatic HCC were compared,and differential lectin affinitied glycans were further verified by multi-lectin blotting.Measurement data were analyzed by two-sample heteroscedastic t test.Results Both lectin microarray and lectin boltting assay showed enhanced affinity on wheat germ agglutintin（WGA）,sambucus nigra lectin（SNA）,phaseolus vulgaris erythroagglutinin（PHAE） and datura stramonium lectin（DSA） in metastatic HCC serum proteins（P0.05）.It showed that the specifc glycan profiles of metastatic HCC serum glycoprotein involved increased Sia,GlcNAc,bisecting GlcNac,multi-antennary and biantennary N-glycan structure. Conclusions Our study indicated that the alternated sugar chains might be associated with tumor invasion and metastasis.The potential value of these specific glycan aberrations as HCC metastasis biomarkers worth further studying.

关 键 词：凝集素芯片 肝癌 肿瘤转移 聚糖 

分 类 号：R735.7[医药卫生—肿瘤]