利用基因芯片分析孕妇外周血浆APC基因甲基化初步探究.docVIP

利用基因芯片分析孕妇外周血浆APC基因甲基化初步探究 　 作者:黄钧，葛芹玉，蒋犁，蒋小青，陆祖宏 【摘要】 目的 研究孕妇外周血浆中APC基因甲基化模式与孕妇妊娠状态的关系，探讨其应用于无创性产前诊断的可行性。方法 从不同妊娠时期孕妇和健康未孕妇女外周血浆标本中提取APC基因，经过亚硫酸氢钠转化后，利用基因芯片方法检测分析APC基因启动区一组CpG位点的甲基化模式，并对不同妊娠时期以及健康未孕样品进行了分析比较。结果 健康未孕妇女血浆中APC基因是未甲基化或低甲基化的，而在妊娠状态下是高甲基化状态的，但不同妊娠时期甲基化程度有所不同。结论 孕妇外周血浆APC甲基化模式的变化与妊娠状态和时间均有一定的关系，基因芯片方法可以高通量的检测分析血浆DNA甲基化状态。血浆DNA甲基化的分析进一步应用于无创性产前诊断具有可行性。 【关键词】 无创性产前诊断；DNA甲基化；基因芯片；APC基因 A microarray to analyze methylation pattern of the APC gene in maternal plasma in pregnant woman 【Abstract】 Objective Detection of the relationship between methylation pattern of the APC gene in maternal plasma in pregnant woman and the state of pregnancy， and estimation the potentiality that applicate to noninvasive prenatal diagnosis. Methods Plasma DNA was isolated from peripheral blood of pregnant women， then after bisulphate-treat， the methylation pattern of CpG site in the APC promoter were analyzed by DNA microarray-based method. The methylation pattern of differential stages in pregnant women and nonpregnant women were compared by DNA microarray. Results The methylation prattern of CpG site in the APC promoter in healthy nonpregnant women are densely methylation and in pregnant women are hypomethylation or unmethylation. The degree of methylation in pregnant women are distinction. Conclusion There are relationship between the methylation pattern of the APC gene in maternal plasma in pregnant women and the pattern of the differential stages and times of pregnancy. The DNA microarray can be applied as a high throughput tool to map methylation patterns in CpG island sites. The analysis of DNA methylation in plasma will have the powerful potentiality that applicate to noninvasive prenatal diagnosis. 【Key words】 DNA methylation; noninvasive prenatal diagnosis;gene microarray; APC gene 1997年，Lo等［1］首次报道在母体血循环中发现了游离胎儿DNA，证实了母体血浆中的DNA是胎儿和母体DNA的混合体，这一发现为无创性产前诊断开辟了新的方法。但以往对孕妇外周血中游离胎儿DNA的应用大多局限在检测胎儿从父亲一方继承来的基因或突变，且检测的胎儿基因必须能与母亲DNA 序列显著区分开来。2002 年， Poon等［2］利用后生性标志，如胎儿