Gene expression profiling of human endometrial receptivity on days LH+2 versus LH+7 by microarray technology
Gene expression profiling of human endometrial receptivity on days LH+2 versus LH+7 by microarray technology
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DOI：
10.1093/molehr/gag037
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发表时间：
2003-05-01
影响因子：
4
通讯作者：
Simón, C
中科院分区：
文献类型：
--
作者：
Riesewijk, A;Martín, J;Simón, C
在人类中，胚胎着床和繁殖依赖于胚胎与接受子宫内膜的相互作用。为了获得对人类子宫内膜容受性的全面分子理解，我们在五个独立的实验中比较了同一名有生育能力的妇女（n = 5）在同一月经周期中获得的接受前（LH+ 2天）和接受期（LH+7）子宫内膜的基因表达谱。使用Affymetrix HG-U95A阵列对活组织切片进行分析，这是一种包含类似12,000个基因的DNA芯片。使用预先定义的标准，即至少五分之四的女性的倍数变化大于或等于3，我们确定了211个受调节的基因。其中，153个在LH+7和LH+2位点上调，58个下调。在这211个受调控的基因中，我们确定了已知在接受性子宫内膜发育中起作用的基因，以及在子宫内膜接受性甚至子宫内膜表达中起作用的基因，这些基因以前没有被描述过。通过实时荧光定量PCR （Q-PCR）对来自育龄妇女（n = 3）独立的LH+2和LH+7子宫内膜样本（n = 3）和整个月经周期（n = 15）中三个上调基因[谷胱甘肽过氧化物酶3 (GPx-3), claudin-4 （claudin-4）和溶质载体家族1成员1 (SLC1A1)]的mRNA进行定量分析，验证阵列数据。人claudin-4在植入期达到峰值，而GPx-3和SLC1A1在分泌后期表达最高。原位杂交（ISH）实验表明，GPx-3和SLC1A1的表达在黄体中晚期仅限于腺和腔上皮细胞。本工作为该领域增加了新的重要数据，并强调了即使使用全局基因表达分析研究子宫内膜容受性的复杂性。
In humans, embryonic implantation and reproduction depends on the interaction of the embryo with the receptive endometrium. To gain a global molecular understanding of human endometrial receptivity, we compared gene expression profiles of pre-receptive (day LH+2) versus receptive (LH+7) endometria obtained from the same fertile woman (n = 5) in the same menstrual cycle in five independent experiments. Biopsies were analysed using the Affymetrix HG-U95A array, a DNA chip containing similar to12 000 genes. Using the pre-defined criteria of a fold change greater than or equal to3 in at least four out of five women, we identified 211 regulated genes. Of these, 153 were up-regulated at LH+7 versus LH+2, whereas 58 were down-regulated. Amongst these 211 regulated genes, we identified genes that were known to play a role in the development of a receptive endometrium, and genes for which a role in endometrial receptivity, or even endometrial expression, has not been previously described. Validation of array data was accomplished by mRNA quantification by real time quantitative fluorescent PCR (Q-PCR) of three up-regulated [glutathione peroxidase 3 (GPx-3), claudin 4 (claudin-4) and solute carrier family 1 member 1 (SLC1A1)] genes in independent LH+2 versus LH+7 endometrial samples from fertile women (n = 3) and the three up-regulated genes throughout the menstrual cycle (n = 15). Human claudin-4 peaks specifically during the implantation window, whereas GPx-3 and SLC1A1 showed highest expression in the late secretory phase. In-situ hybridization (ISH) experiments showed that GPx-3 and SLC1A1 expression was restricted to glandular and luminal epithelial cells during the mid- and late luteal phase. The present work adds new and important data in this field, and highlights the complexity of studying endometrial receptivity even using global gene-expression analysis.