CAJ | 학술논문

目的建立骨髓增生异常综合征(MDS)的CpG岛甲基化模式,为MDS的早期诊断和鉴别诊断提供新的生物学标志.方法采用Illumina450k甲基化芯片和Agilent全基因组表达谱基因芯片进行差异基因筛选,筛选出高甲基化低表达基因,初步建立MDS的CpG岛甲基化模式.通过甲基化特异性PCR、重硫酸盐测序和实时荧光定量PCR进一步在MDS患者组及对照组(非恶性血液病患者)中进行验证,最终建立CpG岛甲基化模式.最后采用诊断试验评价的方法评价CpG岛甲基化模式诊断MDS的效能.结果通过甲基化芯片和表达谱基因芯片的关联分析,以及在211例MDS患者组和60例对照组中进行验证,建立的MDS CpG岛甲基化模式为高甲基化低表达的6个基因.6个基因在MDS组发生甲基化的比例分别为ABAT (97％)、DAPP1 (98％)、FADD(89％)、LRRFIP1 (96％)、PLBD1 (89％)和SMPD3 (85％).5个或5个以上基因同时发生甲基化时诊断MDS的特异度和灵敏度为95.0％和91.4％,准确度为92.3％.结论由ABAT、DAPP1、FADD、LRRFIP1、PLBD1、SMPD3这6个基因建立的MDS的CpG岛甲基化模式可以提高MDS诊断的准确性及鉴别诊断效能.
목적 건립골수증생이상종합정(MDS)적CpG도갑기화모식,위MDS적조기진단화감별진단제공신적생물학표지.방법 채용Illumina450k갑기화심편화Agilent전기인조표체보기인심편진행차이기인사선,사선출고갑기화저표체기인,초보건립MDS적CpG도갑기화모식.통과갑기화특이성PCR、중류산염측서화실시형광정량PCR진일보재MDS환자조급대조조(비악성혈액병환자)중진행험증,최종건립CpG도갑기화모식.최후채용진단시험평개적방법평개CpG도갑기화모식진단MDS적효능.결과 통과갑기화심편화표체보기인심편적관련분석,이급재211례MDS환자조화60례대조조중진행험증,건립적MDS CpG도갑기화모식위고갑기화저표체적6개기인.6개기인재MDS조발생갑기화적비례분별위ABAT (97％)、DAPP1 (98％)、FADD(89％)、LRRFIP1 (96％)、PLBD1 (89％)화SMPD3 (85％).5개혹5개이상기인동시발생갑기화시진단MDS적특이도화령민도위95.0％화91.4％,준학도위92.3％.결론 유ABAT、DAPP1、FADD、LRRFIP1、PLBD1、SMPD3저6개기인건립적MDS적CpG도갑기화모식가이제고MDS진단적준학성급감별진단효능.
Objective To identify methylation profiles in myelodysplastic syndrome (MDS) and to provide the biomarkers for the early diagnosis and differential diagnosis of MDS.Methods Genes were screened for hypermethylation by genome-wide DNA methylation profiles.Transcription down-regulation was determined with a gene expression microarray.Methylation-specific,real-time,and bisulfitesequencing PCR cloning and sequencing were performed to validate selected genes in MDS cases and nonmalignant hematologic diseases (controls).Diagnostic test,such as sensitivity and specificity,was used to evaluate the value of methylation patterns.Results A draft of methylation patterns was established and refined to 6 genes after validation in 211 patients and 60 controls.The hypermethylated genes were ABAT (97％),DAPP1 (98％),FADD (89％),LRRFIP1 (96％),PLBD1 (89％),and SMPD3 (85％).A combination of 5 or more than 5 genes showed a specificity of 95％ and sensitivity of 91.4％ for the diagnosis of MDS.The accuracy of diagnosis was 92.3％.Conclusions We demonstrated here that the ABAT,DAPP1,FADD,LRRFIP1,PLBD1 and SMPD3 genes are hypermethylated and downregulated in MDS.The six genes could be the markers of the methylation patterns in MDS,as a noninvasive approach for the diagnosis of MDS.