H19基因甲基化在Y染色体异常男性不育患者中的表达  

H19 gene methylation levels in male infertile patients with Y chromosome abnormality

作　　者：陈贺[1] 贺文风[1] 牟海燕[1] 匡渤海[2] CHEN He HE Wen-feng MU Hai-yan KUANG Bo-hai(The Key Laboratory of Molecular Medicine, The Second Affiliated Hospital of Nanchang University, Nanchang 330006, Jiangxi Department of Medical Genetics, Medical School of Nanchang University, Nanchang 330006, Jiangxi , China)

机构地区：[1]南昌大学第二附属医院江西省分子医学重点实验室,南昌330006 [2]南昌大学医学院医学遗传研究室,南昌330006

出　　处：《临床检验杂志》2016年第11期861-864,共4页Chinese Journal of Clinical Laboratory Science

基　　金：江西省卫生厅科技计划项目(20165290)

摘　　要：目的探讨H19基因甲基化在Y染色体异常男性不育患者中的表达及相关性。方法用实时荧光PCR探针法检测33例Y染色体异常男性不育患者的微缺失。梯度离心法提取33例Y染色体异常标本,采用亚硫酸氢盐对精液DNA进行修饰,修饰后的DNA样本进行PCR扩增及纯化,产物与p MD@18-T载体连接并经酶切验证,挑选阳性克隆测序并对H19基因甲基化程度进行差异分析。结果基因微缺失检测结果显示,33例Y染色体异常患者包括3例AZFa区缺失患者、14例AZFc区缺失患者、1例AZFbc区缺失患者和15例无缺失患者。33例患者中18例Y染色体微缺失患者的H19基因呈低甲基化水平;15例无缺失患者中有7例呈高甲基化水平,其余8例为正常甲基化水平。结论 AZF缺失男性不育患者与H19基因异常甲基化有关,在精子的生成和成熟过程起到重要作用。Objective To investigate the levels of H19 gene methylation in male infertility patients with Y chromosome abnormality.Methods The microdeletions in 33 patients with Y chromosome abnormality were detected by real-time PCR. Spermatozoa were extracted by gradient centrifugation from 33 semen samples,and sperm DNAs were modified with hydrosulfite. The modified DNAs were amplified by PCR and purified. The purified PCR products were ligated with p MD@ 18-T vector and then verified by restriction endonuclease digestion. The positive clones were performed sequencing,and the levels of H19 gene methylation were analyzed. Results The microdeletion results in 33 patients with Y chromosome abnormality showed 3 with AZFa deletion,14 with AZFc deletion,1 with AZFbc deletion and 15 without any deletion. Eighteen patients with Y chromosome microdeletion had low methylation levels of H19 gene.Among 15 patients without any deletion,7 had high methylation levels of H19 gene,and the other 8 had normal methylation levels.Conclusion Male infertility patients with AZF deletion are associated with abnormal methylation of H19 gene,which may play an important role in the process of sperm production and maturation.

关 键 词：H19基因甲基化 Y染色体微缺失 男性不育 相关性 

分 类 号：R698.2[医药卫生—泌尿科学]