分析化学2012，Vol.40Issue(8)：1201-1206,6.DOI:10.3724/SP.J.1096.2012.11255
液相色谱-串联质谱法测定生物样本全基因组DNA甲基化
Determination of Global DNA Methylation in Biological Samples by Liquid Chromatography- Tandem Mass Spectrometry
摘要
Abstract
A method based on liquid chromatography-electrospray ionization tandem mass spectrome-try (LC-ESI-MS/MS) was developed to determine global DNA methylation level in biological samples. DNA was extracted from biological samples and digested by three enzymes into single nucleo-tides. Liquid chromatography coupled to tandem mass spectrometry was used to measure the concentrations of 2 -deoxycytidine and 2'-deoxy-5-methylcytidine respectively, so as to calculate the global DNA methylation ratios. The developed method was further used to explore the global DNA methylation level in normal human liver cell L-02 exposed to perfluorooctane sulfonate (PFOS) and plasma samples from 10 hepatocellular carcinoma patients and 10 control cases. This approach has high sensitivity and stability, and is easy to operate, enabling us to analyze the global DNA methylation level in various biological samples, especially those valuable samples (such as serum, plasma etc.) with extremely low concentration of DNA.关键词
DNA甲基化/液相色谱/质谱/血浆/细胞/全氟辛烷磺酸Key words
Deoxyribonucleic acid methylation/ Liquid chromatography/ Mass spectrometry/ Plasma/ Cell/ Perfluorooctane sulfonate引用本文复制引用
彭思远,张洁,田美平,王展林,申河清..液相色谱-串联质谱法测定生物样本全基因组DNA甲基化[J].分析化学,2012,40(8):1201-1206,6.基金项目
本文系中国科学院知识创新工程重要方向项目(No.KZCX2-EW-QN408)、“百人计划”项目和厦门市科技项目(No.3502Z20112017)资助 （No.KZCX2-EW-QN408）
