Quantification of 5-methylcytosine, 5-hydroxymethylcytosine and 5-carboxylcytosine from the blood of cancer patients by an enzyme-based immunoassay.
Quantification of 5-methylcytosine, 5-hydroxymethylcytosine and 5-carboxylcytosine from the blood of cancer patients by an enzyme-based immunoassay.
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DOI：
10.1016/j.aca.2014.09.020
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发表时间：
2014-12-10
影响因子：
6.2
通讯作者：
Irudayaraj J
中科院分区：
文献类型：
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作者：
Chowdhury B;Cho IH;Hahn N;Irudayaraj J
经典表观遗传修饰5-甲基胞嘧啶（5 mC）的全基因组畸变被认为是基因沉默的标志，在介导健康细胞的致癌转化中起着关键作用。近年来，在哺乳动物基因组中发现了3种由5 mC酶促氧化而衍生的表观遗传标记，即5-羟甲基胞嘧啶（5 hmC）、5-甲酰基胞嘧啶（5 fC）和5-羧基胞嘧啶（5caC）。越来越多的证据表明，这些新的碱基具有独特的调节功能，并可能在致癌作用中发挥关键作用。为了给这些罕见的表观遗传标记提供定量基础，我们设计了一种生物素-亲和素介导的酶免疫分析（EIA），并评估了其在从诊断为转移性肺癌、胰腺癌和膀胱癌患者以及健康对照的血液中分离的基因组DNA中的性能。建议EIA采用空间优化的生物素化抗体和高度的辣根过氧化物酶（HRP）标记的链霉亲和素，促进信号放大和灵敏的检测。结果表明，正常人血液基因组DNA中5 mC、5 hmC和5caC的含量分别为1.025 ± 0.081、0.023 ± 0.006和0.001 ± 0.0002。我们观察到与健康对照相比，恶性肺癌患者血液中5 hmC的平均总体百分比（0.013 ± 0.003%）显著（p<0.05）降低。这些表观遗传修饰在癌症中的精确生物学作用仍不清楚，但在过去两年中，很明显，在各种癌症中，全球5 hmC含量急剧减少。据我们所知，这是转移性肺癌患者血液中5 hmC含量降低的第一份报告，这一观察结果的临床效用需要在更大的样本数据集中进一步验证。
Genome-wide aberrations of the classic epigenetic modification 5-methylcytosine (5mC), considered the hallmark of gene silencing, has been implicated to play a pivotal role in mediating carcinogenic transformation of healthy cells. Recently, three epigenetic marks derived from enzymatic oxidization of 5mC namely 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC) and 5-carboxylcytosine (5caC), have been discovered in the mammalian genome. Growing evidence suggests that these novel bases possess unique regulatory functions and may play critical roles in carcinogenesis. To provide a quantitative basis for these rare epigenetic marks, we have designed a biotin-avidin mediated Enzyme-based Immunoassay (EIA) and evaluated its performance in genomic DNA isolated from blood of patients diagnosed with metastatic forms of lung, pancreatic and bladder cancer, as well as healthy controls. The proposed EIA incorporates spatially optimized biotinylated antibody and a high degree of horseradish-peroxidase (HRP) labeled streptavidin, facilitating signal amplification and sensitive detection. We report that the percentages of 5mC, 5hmC and 5caC present in the genomic DNA of blood in healthy controls as 1.025 + 0.081, 0.023 + 0.006 and 0.001 + 0.0002 respectively. We observed a significant (p<0.05) decrease in the mean global percentage of 5hmC in blood of patients with malignant lung cancer (0.013 + 0.003 %) in comparison to healthy controls. The precise biological roles of these epigenetic modifications in cancers are still unknown but in the past two years it has become evident that the global 5hmC content is drastically reduced in a variety of cancers. To the best of our knowledge, this is the first report of decreased 5hmC content in the blood of metastatic lung cancer patients and the clinical utility of this observation needs to be further validated in larger sample datasets.
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影响因子：
10.3
作者：
Esteller, M;Silva, JM;Herman, JG
通讯作者：
Herman, JG
影响因子：
11.2
作者：
Jin SG;Jiang Y;Qiu R;Rauch TA;Wang Y;Schackert G;Krex D;Lu Q;Pfeifer GP
通讯作者：
Pfeifer GP
影响因子：
3.4
作者：
BARNARD, DL;JOHNSON, FB;RICHARDS, DF
通讯作者：
RICHARDS, DF
全基因组DNA甲基化分析的进展。
影响因子：
2.7
作者：
Gupta R;Nagarajan A;Wajapeyee N
通讯作者：
Wajapeyee N
影响因子：
64.8
作者：
通讯作者：
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