To quantitatively detect hypermethylation and to analyze methylation pattern, a methylation-specific oligonucleotide microarray for quantitative analysis was developed. The method used bisulfite-modified DNA as a temp...To quantitatively detect hypermethylation and to analyze methylation pattern, a methylation-specific oligonucleotide microarray for quantitative analysis was developed. The 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. The amplified product, therefore, may contain a pool of DNA fragments with altered nucleotide sequences due to differential methylation status. A test sample was hybridized to a set of oligonucleotide arrays that discriminate methylated and unmethylated cytosine at specific nucleotide positions,and quantitative differences in hybridization were determined by fluorescence analysis. Four clinical samples of gastric cancer were quantitatively detected and methylation pattern of five methylated clones were analyzed with the methylation-specific oligonucleotide microarray. The results of microarray hybridization were in agreement with bisulfite genomic DNA sequencing. It showed that methylation-specific oligonucleotide microarray for quantitative analysis is a promising technique for mapping methylation changes in multiple CpG island loci and for generating epigenetic profiles in cancer.展开更多
肺癌是严重威胁人类健康的恶性肿瘤之一，肺癌的早期诊断对其治疗效果十分重要。p16基因是一种重要的抑癌基因，研究肺癌中p16基因启动子区CpG岛甲基化的发生状态具有很大的临床意义。用于检测CpG岛甲基化的特异聚合酶链反应（methylati...肺癌是严重威胁人类健康的恶性肿瘤之一，肺癌的早期诊断对其治疗效果十分重要。p16基因是一种重要的抑癌基因，研究肺癌中p16基因启动子区CpG岛甲基化的发生状态具有很大的临床意义。用于检测CpG岛甲基化的特异聚合酶链反应（methylation specific PCR，MSP）方法，为DNA甲基化的研究提供了新手段。本文采用MSP技术检测肺癌患者全血中p16基因的甲基化情况，探讨其与肺癌发生发展的关系和作为肺癌早期诊断指标的可能性，现报道如下：展开更多
文摘To quantitatively detect hypermethylation and to analyze methylation pattern, a methylation-specific oligonucleotide microarray for quantitative analysis was developed. The 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. The amplified product, therefore, may contain a pool of DNA fragments with altered nucleotide sequences due to differential methylation status. A test sample was hybridized to a set of oligonucleotide arrays that discriminate methylated and unmethylated cytosine at specific nucleotide positions,and quantitative differences in hybridization were determined by fluorescence analysis. Four clinical samples of gastric cancer were quantitatively detected and methylation pattern of five methylated clones were analyzed with the methylation-specific oligonucleotide microarray. The results of microarray hybridization were in agreement with bisulfite genomic DNA sequencing. It showed that methylation-specific oligonucleotide microarray for quantitative analysis is a promising technique for mapping methylation changes in multiple CpG island loci and for generating epigenetic profiles in cancer.
文摘肺癌是严重威胁人类健康的恶性肿瘤之一，肺癌的早期诊断对其治疗效果十分重要。p16基因是一种重要的抑癌基因，研究肺癌中p16基因启动子区CpG岛甲基化的发生状态具有很大的临床意义。用于检测CpG岛甲基化的特异聚合酶链反应（methylation specific PCR，MSP）方法，为DNA甲基化的研究提供了新手段。本文采用MSP技术检测肺癌患者全血中p16基因的甲基化情况，探讨其与肺癌发生发展的关系和作为肺癌早期诊断指标的可能性，现报道如下：
