Russell-Silver综合征11p15.5区域遗传缺陷分析  

Analysis of genetic defects in the 11p15.5 region in Russell-Silver syndrome

作　　者：夏超然[1,2] 杨永臣[3] 许无恨[3] 路兆宁[1] 王伟[2,4] XIA Chaoran;YANG Yongchen;XU Wuhen;LU Zhaoning;WANG Wei(Shanghai Institute of Medical Genetics, Shanghai Children’s Hospital, Shanghai Jiao Tong University ,Shanghai 200040, China;Key Lab of Medical Embryo Molecular Biology, Ministry of Health, and Shanghai Lab of Embryo and Reproduction Engineering,Shanghai 200040, China;Department of Laboratory Medicine, Shanghai Children's Hospital, Shanghai Jiao Tong University,Shanghai 200040, China;Department of Medical Genetics, Shanghai Children's Hospital,Shanghai 200040,China)

机构地区：[1]上海交通大学附属儿童医院,上海市儿童医院,上海交通大学医学遗传研究所,上海200040 [2]卫生部医学胚胎分子生物学重点实验室,上海市胚胎与生殖工程重点实验室,上海200040 [3]上海市儿童医院检验科,上海200040 [4]上海市儿童医院医学遗传科,上海200040

出　　处：《临床儿科杂志》2018年第3期210-215,共6页Journal of Clinical Pediatrics

基　　金：上海重大出生缺陷的筛查、诊断和防治的体系建立(No.2013ZYJB0015)

摘　　要：目的探讨Russell-Silver综合征(RSS)发病机制。方法采集6例男性,年龄6~8岁的临床表型疑似RSS患儿,以及2例患儿父母、5例健康男性儿童的外周血2 m L,分离单个核细胞并提取基因组DNA,应用焦磷酸测序技术进行分析,检测染色体11p15.5上印记基因控制区域(ICR)1的H19基因的甲基化水平。应用甲基化特异性多重连接探针扩增技术(MS-MLPA)对1例焦磷酸测序结果阳性且为RSS患儿的甲基化水平进行验证分析并对相应区域的基因拷贝数进行检测。结果焦磷酸测序结果显示,6例患儿在H19-差异甲基化区域(DMR)的6个Cp G位点的甲基化率为11%~29%;患儿父母及正常对照组对应位点的甲基化率为44%~59%。焦磷酸测序结果阳性的1例患儿对应的MS-MLPA结果显示,H19基因的4个位点甲基化率在10%左右,明显低于正常水平。KCNQ1OT1基因的4个位点甲基化率约为50%,在正常范围内。所测样本的基因拷贝数均在正常范围内。结论 RSS患儿的ICR1的H19-DMR存在甲基化水平异常。Objective To explore the pathogenesis of Russell-Silver syndrome(RSS).Methods Two milliliter peripheral blood samples were collected from6male patients aged6to8years with suspected RSS phenotype,the parents of2patients and5healthy boys.Mononuclear cells were isolated and genomic DNA was extracted.The methylation level of the H19imprinting control region(ICR)1on chromosome11p15.5was detected by pyrosequencing.The methylation status and the copy number variation in the corresponding region of one RSS patient with positive results by pyrosequencing were analysed by methylation-specific multiplex-ligation-dependent probe amplification assay(MS-MLPA).Results Pyrosequencing analysis revealed that the methylation rates on the6CpG targeting sites in H19differentially methylated region(DMR)in the6RSS patients were about11%~29%,which were significantly lower than those in their parents and normal controls(44%~59%).The MS-MLPA results of one patient with positive pyrosequencing showed that the methylation rates of4sites in H19-DMR were about10%,which was obviously lower than the normal level.The methylation rates of the4sites in KCNQ1OT1gene were about50%,which was in the normal range.The copy number variations from all samples detected were in the normal range.Conclusion There is methylation aberration of H19-DMR in ICR1in children with RSS.

关 键 词：Russell-Silver综合征 甲基化异常 11p15.5 基因组印记 

分 类 号：R725.9[医药卫生—儿科]