基于PCR技术的DNA甲基化检测方法研究进展
李克勇;肖武生;常秀丽;吴庆;
摘要(Abstract)：
DNA甲基化是表观遗传修饰最基本的方式,在维持正常细胞功能、胚胎发育和肿瘤发生中起着重要的作用,是当前学术研究的重点和热点。随着研究的深入,为满足不同研究目的需求,DNA甲基化检测方法也得到了较快的发展。本文针对近年来基于PCR技术的DNA甲基化检测方法进行简要的分析总结,为选择适合研究目的的检测方法提供帮助。
关键词(KeyWords)： 表观遗传;DNA甲基化;PCR;检测
基金项目(Foundation): 国家自然科学基金(30671728,30630021)
作者(Authors): 李克勇;肖武生;常秀丽;吴庆;
参考文献(References)：
- [1]HERMAN J G,GRAFF J R,MYOHANEN S,et al.Methylation-specific PCR:a novel PCR assay formethylation status of CpG islands[J].Proc Natl Acad SciUSA,1996,93(18):9821-9826.
- [2]GONZALGO M L,JONES P A.Rapid quantitation ofmethylation differences at specific sites using methylation-sensitive single nucleotide primer extension(Ms-SNuPE)[J].Nucleic Acids Res,1997,25(12):2529-2531.
- [3]GONZALGO M L,JONES P A.Quantitative methylationanalysis using methylation-sensitive single-nucleotide primerextension(Ms-SNuPE)[J].Methods,2002,27(2):128-133.
- [4]SUZUKI H,KAWAI J,TAGA C,et al.A PCR-mediatedmethod for cloning spot DNA on restriction landmarkgenomic scanning(RLGS)gel[J].DNA Res,1994,1(5):245-250.
- [5]XIONG Z,LAIRD P W.COBRA:a sensitive andquantitative DNA methylation assay[J].Nucleic Acids Res,1997,25(12):2532-2534.
- [6]BRENA R M,AUER H,KORNACKER K,et al.Accuratequantification of DNA methylation using combined bisulfiterestriction analysis coupled with the Agilent 2100Bioanalyzer Platform[J].Nucleic Acids Res,2006,34(3):e17.
- [7]BRENA R M,HUANG T H,PLASS C.Quantitativeassessment of DNA methylation:Potential applications fordisease diagnosis,classification,and prognosis in clinicalsettings[J].J Mol Med,2006,84(5):365-377.
- [8]ZESCHNIGK M,BOHRINGER S,PRICE E A,et al.Anovel real-time PCR assay for quantitative analysis ofmethylated alleles(QAMA):analysis of the retinoblastomalocus[J].Nucleic Acids Res,2004,32(16):e125.
- [9]EADS C A,DANENBERG K D,KAWAKAMI K,et al.MethyLight:a high-throughput assay to measure DNAmethylation[J].Nucleic Acids Res,2000,28(8):E32.
- [10]THOMASSIN H,KRESS C,GRANGE T.MethylQuant:asensitive method for quantifying methylation of specificcytosines within the genome[J].Nucleic Acids Res,2004,32(21):e168.
- [11]YAO Q F,KANG X J,HAO Q L,et al.A quantitativeDNA methylation assay using mismatch hybridization andchemiluminescence[J].Biomed Environ Sci,2005,18(1):48-52.
- [12]JIANG C L,JIN S G,PFEIFER G P.MBD3L1 is atranscriptional repressor that interacts with methyl-CpG-binding protein 2(MBD2)and components of the NuRDcomplex[J].J Biol Chem,2004,279(50):52456-52464.
- [13]RAUCH T,PFEIFER G P.Methylated-CpG island recoveryassay:a new technique for the rapid detection ofmethylated-CpG islands in cancer[J].Lab Invest,2005,85(9):1172-1180.
- [14]GEBHARD C,SCHWARZFISCHER L,PHAM T H,et al.Rapid and sensitive detection of CpG-methylation usingmethyl-binding(MB)-PCR[J].Nucleic Acids Res,2006,34(11):e82.
- [15]HAN W,CAUCHI S,HERMAN J G,et al.DNAmethylation mapping by tag-modified bisulfite genomicsequencing[J].Anal Biochem,2006,355(1):50-61.
- [16]RAMSAHOYE B H.Measurement of genome wide DNAmethylation by reversed-phase high-performance liquidchromatography[J].Methods,2002,27(2):156-161.
- [17]FRAGA M F,URIOL E,BORJA DIEGO L,et al.High-performance capillary electrophoretic method for thequantification of 5-methyl 2-’deoxycytidine in genomicDNA:application to plant,animal and human cancertissues[J].Electrophoresis,2002,23(11):1677-1681.
- [18]YANG I,PARK I Y,JANG S M,et al.Rapidquantification of DNA methylation through dNMP analysisfollowing bisulfite-PCR[J].Nucleic Acids Res,2006,34(8):e61.
- [19]WITTWER C T,REED G H,GUNDRY C N,et al.High-resolution genotyping by amplicon melting analysis usingLCGreen[J].Clin Chem,2003,49(6 Pt 1):853-860.
- [20]WOJDACZ T,KDOBROVIC A.Methylation-sensitive highresolution melting(MS-HRM):a new approach for sensitiveand high-throughput assessment of methylation[J].NucleicAcids Res,2007,35(6):e41.
- [21]HAMADA T,MURASAWA S,ASAHARA T.Simplescreening method for differentially methylated regions of thegenome using a small number of cells[J].Biochem BiophysRes Commun,2007,353(2):275-279.