糖类相关基因芯片的设计与制备  

The design and fabrication of glyco-gene microarrays

作　　者：陈闻天[1] 刘晨[1] 于汉杰[1] 颜桦[1] 王岸潇[1] 陈超[1] 李铮[1] 

机构地区：[1]西北大学生命科学学院,国家微检测系统工程技术中心,西安710069

出　　处：《中国科学：化学》2010年第5期538-545,共8页SCIENTIA SINICA Chimica

基　　金：科技部国际科技合作计划项目(2009DFA32730);教育部高等学校博士学科点专项科研基金(200806970018)资助

摘　　要：随着糖组学的发展,研究表明某些疾病如肿瘤等的发生伴随着体内糖蛋白表面糖链结构的变化,而糖链结构的形成与修饰经过糖基转移酶、糖苷酶和磺基转移酶的参与.本研究从Genebank等数据库中选取人类糖基转移酶基因127条、糖苷酶基因34条和磺基转移酶34条以及管家基因10条.通过对每个基因的mRNA序列进行探针设计,制备出一款糖类相关基因芯片,用于研究糖类相关基因的表达谱,旨在揭示疾病发生与糖类相关基因表达变化的关联性.制备的芯片应用于人肝癌细胞系SMMC-7721与正常肝细胞系Chang’s liver的研究,筛选出差异表达的基因34个,其中上调基因19个、下调基因15个.通过对2个上调基因和2个下调基因进行Realtime-PCR验证,得到了一致的实验结果.Glycosylation is the most common posttranslational modification of proteins.Glycosyltransferases are a ubiquitous group of enzymes that catalyse the transfer of a sugar moiety from an activated sugar donor onto saccharide or non-saccharide acceptors.Glycosyltransferase,glycosidase and sulphotransferase participate in the formation and modification of glycan chains.The growing evidences supported that the glycan structures on the cellular surface happened to change in tumors.In this report,127 glycosyltransferases,34 glycosidases,34 sulfotransferases and 10 house keeping genes from the database of Genebank were selected to design 60-mer oligonucleotide probes according to their mRNA sequences.A kind of glyco-gene microarrays related to glycosylation of proteins was prepared and used to generate the glyco-gene expression profiles with liver normal cell line （Chang＇s liver） and liver cancer cell line （SMMC-7721）.The result showed that there were 34 glycogenes of the differential expression,up-regulation of 19 gene expressions and down-regulation of 15 gene expressions by comparing SMMC-7721 cell with Chang＇s liver cell.The 2 genes （Alg10 and Galnt11） of up-regulation and 2 genes （HMCS and Galnt12） of down-regulation in SMMC-7721 cell were analyzed with the Realtime-PCR in order to confirm the results of the arrays.The verification showed high concordance between the two methods.

关 键 词：基因芯片 糖基转移酶 糖苷酶 磺基转移酶 SMMC-7721肝癌细胞系 Chang’s liver肝细胞系 

分 类 号：R450[医药卫生—治疗学]