国际生殖健康/计划生育杂志

国际生殖健康/计划生育 ›› 2012, Vol. 31 ›› Issue (6): 436-438.

• 论著 • 上一篇    下一篇

人卵巢组织冷冻对印迹基因IGF-2/H19甲基化程度及表达的影响

黄丽丽, 王文军, 李 琳, 李瑞岐, 黄 佳, 杨冬梓   

  1. 510120广州，中山大学孙逸仙纪念医院妇产科
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2012-11-15 发布日期:2012-11-15
  • 通讯作者: 杨冬梓

Effect of Human Ovarian Tissue Cryopreservation on Imprinted Gene IGF-2/H19

HUANG Li-li，WANG Wen-jun，LI Lin，LI Rui-qi，HUANG Jia，YANG Dong-zi   

  1. Department of Obstetrics and Gynecology，Sun Yat-Sen Memorial Hospital，Sun Yat-Sen University，Guangzhou 510120，China
  • Received:1900-01-01 Revised:1900-01-01 Published:2012-11-15 Online:2012-11-15
  • Contact: YANG Dong-zi

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摘要： 目的：探讨程序化冷冻和固相表面玻璃化冷冻对人卵巢组织中印迹基因胰岛素样生长因子2（IGF-2）/H１９（一种非可读框架RNＡ）差异性甲基化区（DMR）CpG岛甲基化程度及两基因表达的影响。方法：15例因其他疾病而手术的患者，将病理检查剩余的卵巢组织切割成皮质小条后随机分配到新鲜组（F组）、程序化冷冻组 （SF组）和固相表面玻璃化组（SSV组）。通过甲基化聚合酶链反应（MS-PCR）检测各组卵巢组织中IGF-2/H19 CpG岛甲基化程度，并分别采用免疫组化法和原位杂交方法检测各组卵泡中IGF-2蛋白和H19 RNA的表达情况。结果：F组、SF组和SSV组卵巢组织甲基化程度分别为26.46%，41.02%和33.09%，虽然SF组和SSV组的甲基化程度略高于F组，但差异无统计学意义。在新鲜和冻融后的卵巢组织中，各级别卵泡的卵母细胞和颗粒细胞胞质中均有IGF-2蛋白阳性表达，3组之间原始和初级卵泡IGF-2的表达差异无统计学意义（P＞0.05）。原始卵泡的颗粒细胞和卵母细胞均可见到H19 RNA阳性表达，3组之间原始卵泡H19 RNA的表达差异无统计学意义（P＞0.05）。结论：未检测到慢速程序化冷冻和快速固相表面玻璃化冷冻对印迹基因IGF-2/H19 CpG岛甲基化程度的影响，亦未发现2种冷冻方法对原始和初级卵泡中IGF-2蛋白和H19 RNA表达的影响。

关键词: 卵巢, 低温保存, 甲基化, 基因, 生殖技术, 辅助

Abstract: Objective：To evaluate the effect of solid-surface vitrification （ＳＳＶ） and slow-freezing on methylation profile of imprinted gene IGF-2/H19 DMR, and its effect on the expression of these two genes in follicles embedding in ovarian tissue. Methods：Human ovarian slices from 15 patients were randomly allocated to Fresh，SF and SSV groups. The methylation profile of imprinted gene IGF-2/H19 was assessed by MS-PCR. The expressions of IGF-2 protein and H19 RNA in follicles embedding in ovarian tissue from different groups were detected by immunohistochemistry and in situ hybridization. Results：The methylation of ovarian tissue in F， SF and SSV group was 26.46%， 41.02% and 33.09%， respectively. The SSV and SF groups did not show significantly difference（P＞0.05）. The expressions of IGF-2 protein and H19 RNA in primordial and primary follicles among F， SF and SSV groups did not show significantly difference （P＞0.05）. Conclusions：There was evidence at present to indicate that cryopreservation of human ovarian tissue had adverse effect on the methylation profile of imprinted gene IGF-2/H19.

Key words: Ovary, Cryopreservation, Methylation, Genes, Reproductive techniques, assisted