Kimura Naoki, Moribe Toyoki, Iizuka Norio, Miura Toshiaki, Tamatsukuri Shigeru, Ishitsuka Hideo, Hamamoto Yoshihiko, Oka Masaaki
Research Group, Molecular Diagnostics R&D Department, Roche Diagnostics K.K., 6-1, Shiba 2-chome, Minato-ku, Tokyo 105-0014, Japan.
Clin Biochem. 2009 Jul;42(10-11):1113-22. doi: 10.1016/j.clinbiochem.2009.03.017. Epub 2009 Mar 28.
To assess the medical applicability of CpG methylation as molecular markers for cancer diagnosis, we established a new system to determine DNA methylation based on TaqMan PCR combined with a methyl-binding-domain polypeptide 2.
We evaluated the diagnostic applicability of this approach by examining the methylation status of two tumor suppressor genes, RASSF1A and APC, in 10 paired hepatocellular carcinoma (HCC) and the corresponding non-tumor liver tissues.
Methylation levels of total 20 clinical samples measured by the TaqMan PCR assay showed a significantly positive correlation (R=0.814, P<0.0005 for RASSF1A, R=0.736, P<0.00001 for APC) with those calculated by bisulfite sequencing. The methylated DNA amount measured by our TaqMan PCR system precisely replicated the methylation status estimated by direct sequencing.
This suggests our method may serve as a reliable and easy-to-use tool for cancer diagnosis using methylated genes as biomarkers.
为评估CpG甲基化作为癌症诊断分子标志物的医学适用性，我们建立了一种基于TaqMan PCR结合甲基结合域多肽2来确定DNA甲基化的新系统。
我们通过检测10对肝细胞癌（HCC）及其相应的非肿瘤肝组织中两个抑癌基因RASSF1A和APC的甲基化状态，评估了该方法的诊断适用性。
通过TaqMan PCR检测的20个临床样本的甲基化水平与亚硫酸氢盐测序计算的结果显示出显著正相关（RASSF1A的R = 0.814，P < 0.0005；APC的R = 0.736，P < 0.00001）。我们的TaqMan PCR系统测量的甲基化DNA量精确地反映了直接测序估计的甲基化状态。
这表明我们的方法可能成为一种可靠且易于使用的工具，用于以甲基化基因作为生物标志物的癌症诊断。
