定量检测E-cadherin基因启动子区甲基化芯片的建立  

Establishment of microarray-based method for quantificationally detecting methylation changes of E-cadherin gene promoter

作　　者：张凡[1] 王燕[2] 陈宝安[1] 郑文莉[2] 杜鹃[1] 陆祖宏[2] 

机构地区：[1]东南大学临床医学院,江苏南京210009 [2]东南大学生物科学与医学工程系,江苏南京210096

出　　处：《东南大学学报（医学版）》2006年第3期182-187,共6页Journal of Southeast University(Medical Science Edition)

基　　金：国家自然科学基金资助项目(30371606;39970832);东南大学科学基金资助项目(XJ0490169)

摘　　要：目的:拟建立一种能够定量检测抑癌基因E-cadherin甲基化改变的基因芯片。方法:用亚硫酸氢盐处理基因组DNA,并以此为模板进行PCR扩增。目的序列中未甲基化的CpG位点被翻转成TpG,而甲基化的CpG位点保持不变。设计5组探针构建一种检测E-cadherin基因启动子区17个CpG位点甲基化改变的芯片,每组探针包括一对非甲基化探针和甲基化探针,检测相邻的3或4个CpG位点。结果:绘制标准曲线显示,芯片上探针的可重复性和精确性很好,并定量检测了1例白血病样本的甲基化改变。结论:基因芯片能够作为一种定量检测基因多个CpG位点甲基化改变的有效工具,并用于肿瘤的研究。Objective To establish a microarray-based method for quantificationally detecting methylation changes of a tumor suppressor gene E-cadherin. Methods This method used bisulfite-modified DNA as a template for PCR amplification, resulting in conversion of unmethylated cytosine, but not methylated cytosine, into thymine within CpG islands of interest. Five sets of oligonucleotide probes were designed to fabricate a DNA microarray to detect 17 CpG sites in the promoter of E- cadherin gene, each set contained a pair of methylated and unmethylated oligonucleotides for the levels of methylation changes detected in a leukemia sample by microarray assay. Conclusion Microarray assay can be applied as a useful tool for mapping methylation changes in multiple CpG loci and for researching cancer.

关 键 词：甲基化 基因芯片 E-CADHERIN基因 

分 类 号：Q78[生物学—分子生物学]