文献类型：期刊文献

英文题名：Highly sensitive detection of DNA methylation levels by using a quantum dot-based FRET method

作者：Ma, Yunfei[1,2];Zhang, Honglian[2];Liu, Fangming[2];Wu, Zhenhua[2];Lu, Shaohua[3];Jin, Qinghui[2];Zhao, Jianlong[2];Zhong, Xinhua[1];Mao, Hongju[2]

机构：[1]E China Univ Sci & Technol, Inst Appl Chem, Shanghai 200237, Peoples R China;[2]Chinese Acad Sci, Shanghai Inst Microsyst & Informat Technol, State Key Lab Transducer Technol, Shanghai 200050, Peoples R China;[3]Fudan Univ, Zhongshan Hosp, Dept Pathol, Shanghai 200032, Peoples R China

年份：2015

卷号：7

期号：41

起止页码：17547

外文期刊名：NANOSCALE

收录：;EI(收录号：20154401460101);WOS:【SCI-EXPANDED(收录号:WOS:000363181600046)】；

基金：This work was supported by Grants from the National Basic Research Program of China (973 Program) (No. 2012CB933303), the National Science Foundation of China (No. 61571429, 61401442, 21275153, 61006086, 21175043 and 21301059), Science and Technology Commission of Shanghai Municipality (Grant No. 12441902600), Shanghai Clinical Center/Shanghai Xuhui Central Hospital, Chinese Academic of Sciences (No. BRC2012002).

语种：英文

外文关键词：Genes - Forster resonance energy transfer - Nanocrystals - Alkylation - Energy transfer - Diseases - Polymerase chain reaction - Semiconductor quantum dots - Diagnosis - DNA

摘要：DNA methylation is the most frequently studied epigenetic modification that is strongly involved in genomic stability and cellular plasticity. Aberrant changes in DNA methylation status are ubiquitous in human cancer and the detection of these changes can be informative for cancer diagnosis. Herein, we reported a facile quantum dot-based (QD-based) fluorescence resonance energy transfer (FRET) technique for the detection of DNA methylation. The method relies on methylation-sensitive restriction enzymes for the differential digestion of genomic DNA based on its methylation status. Digested DNA is then subjected to PCR amplification for the incorporation of Alexa Fluor-647 (A647) fluorophores. DNA methylation levels can be detected qualitatively through gel analysis and quantitatively by the signal amplification from QDs to A647 during FRET. Furthermore, the methylation levels of three tumor suppressor genes, PCDHGB6, HOXA9 and RASSF1A, in 20 lung adenocarcinoma and 20 corresponding adjacent nontumorous tissue (NT) samples were measured to verify the feasibility of the QD-based FRET method and a high sensitivity for cancer detection (up to 90%) was achieved. Our QD-based FRET method is a convenient, continuous and high-throughput method, and is expected to be an alternative for detecting DNA methylation as a biomarker for certain human cancers.