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基于石英晶体微天平测量的无标记实时 DNA 甲基化检测。

Label-free real-time detection of DNA methylation based on quartz crystal microbalance measurement.

机构信息

Division of Nanobiomedicine, Suzhou Institute of Nano-Tech and Nano-Bionics, Chinese Academy of Sciences, Suzhou, Jiangsu 215123, People's Republic of China.

出版信息

Anal Chem. 2013 Feb 19;85(4):2096-101. doi: 10.1021/ac3026724. Epub 2013 Jan 29.

Abstract

DNA methylation plays an important role in the regulation of gene transcription, chromatin compaction, genome imprinting, and X-chromosome inactivation. DNA methyltransferase is considered a potential target for anticancer drug design. It is important to locate aberrantly methylated sequences on the human genome that are linked to specific diseases and to discover new low-toxic methylation inhibitors for medical treatments. We developed a DNA methylation detection method using a quartz crystal microbalance (QCM). We applied this method to assay genes p16 and GALR2 in two cell lines. Methylation of p16 was detected in both HT29 and HepG2 cell lines, whereas methylation of GALR2 was detected only in the HT29 cell line. We also used this method to evaluate the effect of 5-aza-2'-deoxycytidine (decitabine), a methyltransferase inhibitor used in clinical treatment. We found methylation of genes p16 and GALR2 to be strongly inhibited. The results show that this method is sensitive to DNA methylation and is fit for evaluation of methyltransferase inhibitors.

摘要

DNA 甲基化在基因转录调控、染色质紧缩、基因组印记和 X 染色体失活中发挥着重要作用。DNA 甲基转移酶被认为是抗癌药物设计的潜在靶点。重要的是要找到与特定疾病相关的人类基因组上异常甲基化的序列,并发现新的低毒性甲基化抑制剂用于医疗。我们开发了一种使用石英晶体微天平 (QCM) 的 DNA 甲基化检测方法。我们将该方法应用于两种细胞系中的 p16 和 GALR2 基因的测定。在 HT29 和 HepG2 细胞系中均检测到 p16 的甲基化,而仅在 HT29 细胞系中检测到 GALR2 的甲基化。我们还使用该方法评估了甲基转移酶抑制剂 5-氮杂-2'-脱氧胞苷 (地西他滨) 在临床治疗中的作用。我们发现基因 p16 和 GALR2 的甲基化被强烈抑制。结果表明,该方法对 DNA 甲基化敏感,适用于评估甲基转移酶抑制剂。

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