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Epigenome-based cancer risk prediction: rationale, opportunities and challenges.基于表观基因组的癌症风险预测：原理、机遇与挑战。
Nat Rev Clin Oncol. 2018 May;15(5):292-309. doi: 10.1038/nrclinonc.2018.30. Epub 2018 Feb 27.
3
Intratumor heterogeneity in epigenetic patterns.肿瘤内表观遗传模式的异质性。
Semin Cancer Biol. 2018 Aug;51:12-21. doi: 10.1016/j.semcancer.2018.01.010. Epub 2018 Jan 31.
4
Cancer Progenitor Cells: The Result of an Epigenetic Event?癌症祖细胞：一种表观遗传事件的结果？
Anticancer Res. 2018 Jan;38(1):1-6. doi: 10.21873/anticanres.12184.
5
Identification of CpG Sites of Promoter with Opposite Methylation Patterns in Benign and Malignant Prostate Cells.良性和恶性前列腺细胞中具有相反甲基化模式的启动子CpG位点的鉴定
Anticancer Res. 2017 Dec;37(12):6609-6618. doi: 10.21873/anticanres.12118.
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Current insights into functions of phospholipase A2 receptor in normal and cancer cells: More questions than answers.目前对磷脂酶 A2 受体在正常细胞和癌细胞中的功能的了解：问题多于答案。
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7
Assessing alternative base substitutions at primer CpG sites to optimise unbiased PCR amplification of methylated sequences.评估引物CpG位点处的替代碱基替换，以优化甲基化序列的无偏PCR扩增。
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8
Fetal DNA hypermethylation in tight junction pathway is associated with neural tube defects: A genome-wide DNA methylation analysis.紧密连接通路中的胎儿DNA高甲基化与神经管缺陷相关：全基因组DNA甲基化分析
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9
MassARRAY determination of somatic oncogenic mutations in solid tumors: Moving forward to personalized medicine.MassARRAY 法检测实体瘤中的体基因突变：迈向个体化医疗。
Cancer Treat Rev. 2016 Sep;49:57-64. doi: 10.1016/j.ctrv.2016.07.007. Epub 2016 Jul 29.
10
Epigenetic control of phospholipase A2 receptor expression in mammary cancer cells.乳腺癌细胞中磷脂酶A2受体表达的表观遗传调控
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使用等位基因敏感液滴数字聚合酶链反应（EAST-ddPCR）鉴定和定量异质甲基化DNA片段

Identification and Quantification of Heterogeneously-methylated DNA Fragments Using Epiallele-sensitive Droplet Digital Polymerase Chain Reaction (EAST-ddPCR).

作者信息

Menschikowski Mario, Jandeck Carsten, Friedemann Markus, Richter Susan, Thiem Dana, Lange Björn Sönke, Suttorp Meinolf

机构信息

Institute of Clinical Chemistry and Laboratory Medicine, University Hospital "Carl Gustav Carus", Technical University of Dresden, Dresden, Germany

Institute of Clinical Chemistry and Laboratory Medicine, University Hospital "Carl Gustav Carus", Technical University of Dresden, Dresden, Germany.

出版信息

Cancer Genomics Proteomics. 2018 Jul-Aug;15(4):299-312. doi: 10.21873/cgp.20088.

DOI:10.21873/cgp.20088
PMID:29976635
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6070704/
Abstract

BACKGROUND/AIM: DNA methylation plays an important role in the initiation and propagation of carcinogenesis; however, the role of heterogeneously methylated epialleles is currently not well studied, also due to the lack of sensitive, unbiased and high throughput methods. Here, a newly developed droplet digital PCR (ddPCR)-based method was evaluated regarding its ability to quantify such heterogeneously methylated epialleles with sufficient analytical sensitivity and specificity.

MATERIALS AND METHODS

Genomic DNA from blood leukocytes and bone marrow aspirate of an 8-year old male with B-cell acute lymphoblastic leukemia (B-ALL) and from normal and malignant prostate cell lines were analysed using ddPCR.

RESULTS

By using these DNA samples, the specificity of an applied set of fluorescence-labeled probes was demonstrated as a proof of concept.

CONCLUSION

All individual heterogeneously-methylated epialleles were quantifiable by a set of fluorescence-labeled probes with complementary sequences to epialleles in a closed-tube and high-throughput manner. The new method named epiallele-sensitive droplet digital PCR (EAST-ddPCR) may give new insights in the generation and regulation of epialleles and may help in finding new biomarkers for the diagnosis of benign und malignant diseases.

摘要

背景/目的：DNA甲基化在癌症发生的起始和发展过程中发挥着重要作用；然而，由于缺乏灵敏、无偏倚且高通量的方法，目前对异质性甲基化表观等位基因的作用研究较少。在此，我们评估了一种新开发的基于液滴数字PCR（ddPCR）的方法，该方法能够以足够的分析灵敏度和特异性对这种异质性甲基化表观等位基因进行定量。

材料与方法

使用ddPCR分析了一名8岁B细胞急性淋巴细胞白血病（B-ALL）男性患者的血液白细胞和骨髓抽吸物中的基因组DNA，以及正常和恶性前列腺细胞系中的基因组DNA。

结果

通过使用这些DNA样本，作为概念验证，证明了一组应用的荧光标记探针的特异性。

结论

所有个体的异质性甲基化表观等位基因都可以通过一组与表观等位基因具有互补序列的荧光标记探针在封闭管中以高通量方式进行定量。这种名为表观等位基因敏感液滴数字PCR（EAST-ddPCR）的新方法可能会为表观等位基因的产生和调控提供新的见解，并可能有助于寻找用于诊断良性和恶性疾病的新生物标志物。