体外补救成熟法对人卵细胞印记基因DNA甲基化的影响  

The Impact of in vitro Rescue-Maturation on DNA Methylation of Imprinted Genes in Human Oocytes

作　　者：施晓鋆 陈士岭[2] 郑海燕[2] SHI Xiaoyun CHEN Shiling ZHENG Haiyan(Department of Gynecology and Obstetrics, the Second People＇s Hospital of Guiyang, Guiyang 550081, Guizhou, China Reproductive Center, Department of Gynecology and Obstetrics, Nanfang Hospital of Southern Medical University, Guangzhou 510515, Guangdong, China)

机构地区：[1]贵阳市第二人民医院妇产科,贵州贵阳550081 [2]南方医科大学南方医院妇产科生殖中心,广东广州510515

出　　处：《贵州医科大学学报》2016年第10期1137-1140,共4页Journal of Guizhou Medical University

基　　金：国家自然科学基金项目(81460237)

摘　　要：目的:探讨体外补救成熟的人卵细胞印记基因H19和PEG1的DNA甲基化状态,初步评价卵细胞体外补救成熟的表观遗传学风险。方法:选择卵胞浆内单精子显微注射(ICSI)助孕治疗的不孕症患者54例,长方案超排卵取卵术后收集240个未成熟MⅠ期卵细胞,经体外补救培养24 h后,获得130个成熟MⅡ期卵细胞,采用结合重亚硫酸盐限制性酶分析法(COBRA)检测其中62个卵细胞H19基因和68个卵细胞PEG1基因的DNA甲基化状态。结果:23个卵细胞H19基因经bisulfite-PCR扩增成功,成功率37.0%(23/62),其中5个卵细胞(21.7%,5/23)出现H19异常高甲基化模式;30个卵细胞经bisulfite-PCR扩增成功,成功率44.1%(30/68),其中1个卵细胞(3.3%,1/30)出现PEG1异常低甲基化模式。结论:使用体外补救成熟卵细胞可能增加DNA甲基化缺陷的风险。Objective： To investigate the methylation status of H19 DMR and PEG1 DMR in human MII oocytes rescue-matured from M I oocytes,primarily evaluating the epigenetic risk linked to in vitro rescue-maturation. Methods： 240 M Ⅰ oocytes from 54 patients undergoing an ICSI procedure were additionally cultured for 24 hours,and 130 rescue-matured MⅡ oocytes were obtained. The Combined Bisulfite Restriction Analysis（ COBRA） were used to determine the methylation status of H19 DMR in62 oocytes and PEG1 DMR in 68 oocytes. Results： H19 DMR of 23 oocytes proliferated by bisulfitePCR was successful,successful rate was 37. 0%（ 23 /62）,aberrant methylation patterns were found in21. 7% oocytes（ 5 /23） at H19 DMR; 30 oocytes proliferated by bisulfite-PCR was successful,successful rate was 44. 1%（ 30 /68）,and aberrant methylation patterns were found in 3. 3% oocytes（ 1 /30） at PEG1. Conclusion： The use of the MⅡ-rescue oocytes may increase the risk of methylation errors because the imprinting program might be influenced.

关 键 词：印记基因 H19 PEG1 体外补救成熟 卵细胞 甲基化 

分 类 号：R711.6[医药卫生—妇产科学]