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利用多重监测反应模式(MRM),通过对液相色谱及质谱条件的优化,建立了人体结肠癌组织样品中基因组DNA甲基化的高效液相色谱-电喷雾质谱(HPLC-ESIMS/MS)检测方法,对结肠癌组织及癌旁组织中基因组DNA甲基化进行了检测。结果表明,5-甲基脱氧胞苷(5mdC)的检出限是1pg,线性范围为0.0375～0.5mg/L,工作曲线相关系数大于0.9990,该方法的相对标准偏差为2.32%～9.21%。结肠癌病人样品检测结果表明,结肠癌组织基因组DNA甲基化水平低于相应的癌旁组织(P<0.05)。
Using multiplex monitoring reaction mode (MRM), the HPLC-ESIMS / MS detection of genomic DNA methylation in human colon cancer tissue samples was established by optimizing the conditions of liquid chromatography and mass spectrometry. METHODS: Genomic DNA methylation in colon cancer tissues and adjacent tissues was examined. The results showed that the detection limit of 5-methyldeoxycytidine (5mdC) was 1pg, the linear range was 0.0375-0.5mg / L, the correlation coefficient of working curve was greater than 0.9990, and the relative standard deviation was 2.32% -9.21%. The results of colon cancer patient samples showed that the methylation level of genomic DNA in colon cancer tissues was lower than that in corresponding paracancerous tissues (P <0.05).