Determination of DNA Methylation and Hydroxymethylation Levels in Biological Samples by Field-Amplified Sample Injection-Capillary Zone Electrophoresis with UV Detection

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• 作者：Jing Zhang ; Aimin Sun ; Yi Yang ; Jielan Hu ; Ling Wei ; Bo Gao ; Xueqin Ding&#8230
• 关键词：Field ; amplified sample injection ; Capillary zone electrophoresis ; DNA methylation ; DNA hydroxyl methylation ; Cytosine (C) ; 5 ; Methylcytosine (5 ; mC) ; 5 ; Hydroxymethylcytosine (5 ; hmC)
• 刊名：Chromatographia
• 出版年：2016
• 出版时间：December 2016
• 年：2016
• 卷：79
• 期：23-24
• 页码：1649-1658
• 全文大小：
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chromatography; Proteomics; Pharmacy; Laboratory Medicine;
• 出版者：Springer Berlin Heidelberg
• ISSN：1612-1112
• 卷排序：79

文摘

In this study, a method of field-amplified sample injection coupled with capillary zone electrophoresis with ultraviolet detection was established for evaluation of DNA methylation and hydroxymethylation levels in biological materials. By modifying an existing method, the separation of cytosine (C), 5-methylcytosine (5-mC) and 5-hydroxymethylcytosine (5-hmC) was performed on an uncoated capillary column (40 cm × 75 μm I.D.) using 300 mmol L−1 tris solution (pH 2.90) as running buffer and detected at 280 nm. The detection limits (S/N = 3) of the method were 0.004 ng mL−1 for cytosine (C), 0.01 ng mL−1 for 5-methylcytosine (5-mC), and 0.02 ng mL−1 for 5-hydroxymethylcytosine (5-hmC). The proposed method has been successfully applied to the evaluation of DNA methylation and hydroxymethylation levels of blood samples from 15 hepatocellular carcinoma patients and 5 liver cirrhosis patients and liver tissues from 50 pairs of tumor and matched tumor-adjacent samples.