Chen Bao-an, Zhang Fan, Wang Yan, Zheng Wen-li, Du Juan, Gao Chong, Ding Jia-hua, Sun Yun-yu, Cheng Jian, Wang Jun, Zhao Gang, Chen Ning-na, Lu Zu-hong
The Affiliated Zhongda Hospital of Southeast University, Nanjing 210009, China.
Zhonghua Zhong Liu Za Zhi. 2007 Jan;29(1):41-4.
To quantitatively detect the methylation of E-cadherin gene 5'-CpG islands in acute leukemia by microarray-based DNA analysis and to briefly discuss the role of microarry for detection of methylation in tumors.
Bisulfite-modified DNA was used 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 the methylation changes of E-cadherin gene CpG islands in acute leukemia. By drawing a standard curve to assess the levels of changes in methylation detected in the examined samples.
Microarray assay was successfully used to quantitatively detect methylation changes of E-cadherin gene in 5 acute leukemia samples. Varying degree of methylation was detected in five regions and the hypermethylation region was the same. The result was validated by gene sequencing.
Microarray assay may be applied as an useful tool for mapping methylation changes in multiple CpG loci and for leukemia research. It is more time-saving and labor-saving than gene sequencing and can be used to quantitatively detect changes in methylation with high throughput.
通过基于微阵列的DNA分析定量检测急性白血病中E-钙黏蛋白基因5'-CpG岛的甲基化，并简要探讨微阵列在肿瘤甲基化检测中的作用。
亚硫酸氢盐修饰的DNA用作PCR扩增的模板，导致感兴趣的CpG岛内未甲基化的胞嘧啶而非甲基化的胞嘧啶转化为胸腺嘧啶。设计五组寡核苷酸探针以制备DNA微阵列，用于检测急性白血病中E-钙黏蛋白基因CpG岛的甲基化变化。通过绘制标准曲线来评估检测样本中甲基化变化的水平。
微阵列分析成功用于定量检测5例急性白血病样本中E-钙黏蛋白基因的甲基化变化。在五个区域检测到不同程度的甲基化，且高甲基化区域相同。结果经基因测序验证。
微阵列分析可作为一种有用的工具，用于绘制多个CpG位点的甲基化变化图谱以及白血病研究。它比基因测序更省时省力，可用于高通量定量检测甲基化变化。
