Department of Obstetrics and Gynecology, Faculty of Medicine, Gazi University, 06500 Besevler, Ankara, Turkey.
Eur J Obstet Gynecol Reprod Biol. 2010 Mar;149(1):77-81. doi: 10.1016/j.ejogrb.2009.12.007. Epub 2009 Dec 29.
As genomic imprinting plays a critical role in the development of the placenta, the aim of this study was to detect whether the expression levels of the imprinted genes IGF2 and H19 in the endometrium differ between infertile and fertile women.
Total RNA was extracted from 30 (15 unexplained infertile and 15 fertile) women's endometrial tissue. cDNA was synthesized from total RNAs of each sample. IGF2 and H19 mRNA expression levels were measured quantitatively using the Real Time PCR method. In order to determine the allelic expression of IGF2 and H19, genomic DNA was extracted from endometrial tissues.
When compared with the control group, increased mRNA expression of IGF2 was detected (1.5-fold change, P=0.015) in the unexplained infertility group. In contrast, H19 expression was lower in the infertility group as compared to the control group (4-fold change, P<0.0001). Restriction analysis of cDNA-derived PCR product showed that all patients and controls indicated monoallelic expression of IGF2 and H19.
Our results showed that altered expression of these imprinted genes might affect implantation and that their timely and appropriate activation is important for proper functioning. To understand the molecular epigenetic basis of implantation and placental development, genomic imprinted genes should be further investigated.
由于基因组印迹在胎盘发育中起着关键作用,本研究旨在检测子宫内膜中印迹基因 IGF2 和 H19 的表达水平是否在不孕和生育妇女之间存在差异。
从 30 名(15 名不明原因不孕和 15 名生育)妇女的子宫内膜组织中提取总 RNA。从每个样本的总 RNA 中合成 cDNA。使用实时 PCR 方法定量测量 IGF2 和 H19 mRNA 的表达水平。为了确定 IGF2 和 H19 的等位基因表达,从子宫内膜组织中提取基因组 DNA。
与对照组相比,在不明原因不孕组中检测到 IGF2 mRNA 的表达增加(1.5 倍变化,P=0.015)。相比之下,不孕组中 H19 的表达低于对照组(4 倍变化,P<0.0001)。对 cDNA 衍生 PCR 产物的限制性分析表明,所有患者和对照组均显示 IGF2 和 H19 的单等位基因表达。
我们的结果表明,这些印迹基因的表达改变可能会影响着床,并且它们的及时和适当激活对于正常功能很重要。为了理解着床和胎盘发育的分子表观遗传学基础,应该进一步研究基因组印迹基因。