基于凝集素芯片的不同转移潜能肝癌细胞膜蛋白糖谱比较  

Membrane Protein Glycanprofiling of Hepatocellular Carcinoma Cell With Different Metastastic Potential by Lectin Microarray

作　　者：王念[1,2] 康晓楠[1,2] 刘银坤[1,2] 郭坤[2] 崔杰峰[2] 孙瑞霞[2] 陈洁[2] 赵燕[2] 陈培[1] 

机构地区：[1]复旦大学中山医院肝癌研究所,上海200032 [2]复旦大学生物医学研究院癌症研究中心,上海200032

出　　处：《生物化学与生物物理进展》2009年第10期1348-1355,共8页Progress In Biochemistry and Biophysics

基　　金：国家高技术研究发展计划(863)(2006AA02A308);国家重大传染病防治专项资助项目(2008ZX10002-021,-017)资助项目~~

摘　　要：评估采用凝集素芯片技术寻找肝癌细胞表面侵袭和转移相关特征性糖谱的适用性.首先选取一对模式细胞株(中华仓鼠卵巢细胞CHO和其N-乙酰葡萄糖胺转移酶Ⅰ缺陷株Lec1)验证凝集素芯片系统的可靠性.然后通过凝集素芯片比较正常肝细胞L02、非转移肝癌细胞Hep3B、高转移肝癌细胞HCCLM3的细胞表面糖谱,同时采用细胞凝集素组织化学的方法验证芯片结果.细胞Hep3B和L02相比,对凝集素PHA-L、ConA、AAL、MPL的亲和作用增强而对凝集素WGA的亲和作用减弱,提示在肝癌细胞表面可能出现了增多的复杂寡糖分支、高甘露糖、末端岩藻糖、黏蛋白T抗原和减少的N-乙酰葡萄糖胺和/或多价唾液酸结构.细胞HCCLM3和Hep3B相比,对凝集素LCA、MAL-Ⅰ、MAL-Ⅱ、WGA、PHA-E的亲和作用增强而对凝集素RCA-I的亲和作用减弱,提示在高转移肝癌细胞HCCLM3的表面可能出现了增多的核心岩藻糖、唾液酸(主要是α2-3链接方式)、N-乙酰葡萄糖胺、平分型GlcNAc结构以及减少的末端β1-4链接半乳糖结构.细胞凝集素组织化学的结果支持芯片结果.研究证明,凝集素芯片技术是解析生物学进程中糖谱改变的适用工具.To perform an evaluation of the usefulness of lectin microarray approach for the identification of characteristic glycan profilings related to different aggressive and metastatic potentials between liver cell lines. At first, CHO and its glycosyltransferase-defective mutants Lecl were chose to validate the feasibility of the lectin microarray system, then the difference of the glycan profiling on cell surface of L02 （normal control）, Hep3B （no metastasis） and HCCLM3 （high metastatic potential） cell lines was characterized by lectin mieroarray. Comparing with L02, Hep3B showed increased affinity for PHA-L, ConA, AAL, MPL and decreased signals for WGA. HCCLM3 presented elevated signals for LCA, MAL- I, MAL- II, WGA, PHA-E and decreased signals for RCA-I with contrast to Hep3B. The results of fluoresceinated lectin staining of cells with biotinylated LCA, WGA, PHA-E and RCA-I support some observation from array. Lectin microarray was an applicable and useful tool in identifying the glycan profiling changes that accompany the biological processes.

关 键 词：凝集素芯片 肝癌 糖 转移 

分 类 号：R735.7[医药卫生—肿瘤] Q53[医药卫生—临床医学]