Human endometrium undergoes sequential changes during the menstrual cycle and becomes receptive to implantation during a defined period in the secretory phase. We attempted to identify the genes expressed during this period by representational difference analysis (RDA). When the cDNAs of a proliferative endometrium were used as the driver and the cDNAs of a post-ovulatory day 5 endometrium were used as the tester, a number of bands were identified by RDA. DNA of the cloned RDA products revealed that the majority of the clones contained a fragment of a cDNA identical to that of a crystallin B chain. Northern blot analysis showed that the expression of the alpha crystallin B chain mRNA was absent during the proliferative phase. The expression of the mRNA of alpha crystallin B chain first appeared in the secretory phase, progressively increased during this phase and peaked in the late secretory endometria. The pattern of expression of alpha crystallin B chain mRNA in the endometrium of mature cycling baboons (Papio anubis) was similar to that seen in human endometrium. As revealed by Western blot analysis, the expression of the alpha crystallin B chain protein in human endometrium followed a pattern of expression similar to its mRNA. At the cellular level, the immunoreactive protein first appeared in the surface epithelial cells of human endometrium within the implantation window without significant immunoreactivity in the underlying glandular cells. During the mid- and late secretory phases, the intensity of staining in the epithelial cells was enhanced and an intense immunoreactivity was developed in the glandular epithelium, alpha crystallin B chain was virtually an epithelial product and no immunoreactivity for this protein was detectable in the stromal cells, endothelial cells or lymphoid cells. The expression of alpha crystallin B chain could be regulated, by medroxy progesterone acetate as well as by oestrogen withdrawal, in human endometrial carcinoma cells (EnCa-101), transplanted to nude mice. Based on the data presented here, the known function of alpha crystallin B chain and its distinct pattern of expression in human endometrium, we suggest that this protein is an important factor within the molecular repertoire that makes endometrium receptive to implantation.

译文

人的子宫内膜在月经周期中会发生顺序变化,并在分泌期的特定时期内接受植入。我们试图通过代表性差异分析(RDA)来鉴定在此期间表达的基因。当使用增生性子宫内膜的cDNA作为驱动器,并在排卵后第5天的子宫内膜的cDNA作为检测器时,通过RDA鉴定了许多条带。克隆的RDA产物的DNA显示大多数克隆都包含与crystallin B链相同的cDNA片段。 Northern印迹分析表明在增殖期不存在α晶状体蛋白B链mRNA的表达。 α晶状蛋白B链的mRNA表达首先出现在分泌期,在此阶段逐渐增加,并在晚期分泌性子宫内膜达到高峰。在成熟的狒狒(Papio anubis)的子宫内膜中,α晶状蛋白B链mRNA的表达方式与人的子宫内膜相似。如蛋白质印迹分析所揭示的,人子宫内膜中的α结晶蛋白B链蛋白的表达遵循与其mRNA相似的表达模式。在细胞水平上,免疫反应蛋白首先出现在植入窗口内人子宫内膜的表面上皮细胞中,而在下面的腺体细胞中却没有明显的免疫反应性。在分泌的中期和后期,上皮细胞的染色强度增强,腺上皮形成强烈的免疫反应性,αcrystallin B链实际上是上皮产物,在基质中未检测到该蛋白的免疫反应性细胞,内皮细胞或淋巴样细胞。醋酸甲羟孕酮以及雌激素的撤除均可在移植到裸鼠的人子宫内膜癌细胞(EnCa-101)中调节α晶状蛋白B链的表达。根据此处提供的数据,α晶体蛋白B链的已知功能及其在人子宫内膜中的独特表达模式,我们建议该蛋白是分子库中的一个重要因素,可使子宫内膜易于接受植入。

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