Division of Infertility and Reproductive Endocrinology, Department of Gynecology, Obstetrics and Gynecological Oncology, Poznan University of Medical Sciences, Poznan, Poland.
Department of Cancer Immunology, Poznan University of Medical Sciences, Poznan, Poland.
J Biol Regul Homeost Agents. 2019;33(2):461-468.
Endocrinal interactions are one of the most crucial regulatory mechanisms that maintain the state of homeostasis in humans. Processes such as oogenesis, folliculogenesis, menstruation and pregnancy remain under hormonal control. A key role in folliculogenesis is played by granulosa cells. Moreover, granulosa cells take part in corpus luteum formation after ovulation. Because of that, it is important to understand the ways in which the granulosa cells, associated with those processes, respond to hormonal stimulus. In the present study, a transcriptomic analysis of human granulosa cells (GCs) was carried out with the use of expression microarrays. The results were validated by RT-qPCR. The total RNA was isolated after 1st, 7th, 15th and 30th days of long-term primary cultures. The main focus of this work was placed on the genes belonging to "Response to estradiol", "Response to follicle-stimulating-hormone", "Cellular response to hormone stimulus", "Cellular hormone metabolic process" and "Hormone biosynthetic process" gene ontology groups. These groups of genes have been associated with GC hormone metabolism and cellular response to hormones. Eighty genes belonging to these groups were identified. Those that were members of more than one of the analyzed gene ontology groups, or exhibited unique expression patterns, were selected for further analysis. All of the selected genes were described, with their expression patterns detailed. In this manuscript, two gene expression patterns have been described. The first one showed large downregulation of genes in the later stages of culture, with the second one presenting upregulation of expression after day 1 of IVC. The present research was focused on six genes found to be the most important for steroidogenesis: STAR, POR, CYP11A1, ADM, GCLC, IL1B, as well as three genes of higher expression at the later stages of long-term in vitro culture: NR2F2, BMP4, COL1A1. The main goal of the presented study was to select genes involved in response to hormonal stimulus and hormone metabolism in GC long-term in vitro culture.
内分泌相互作用是维持人体内稳态的最重要的调节机制之一。卵子发生、卵泡发生、月经和妊娠等过程仍然受到激素的控制。在卵泡发生中，颗粒细胞起着关键作用。此外，排卵后颗粒细胞参与黄体形成。因此，了解与这些过程相关的颗粒细胞对激素刺激的反应方式非常重要。在本研究中，使用表达微阵列对人颗粒细胞（GC）进行了转录组分析。结果通过 RT-qPCR 进行了验证。总 RNA 是在长期原代培养的第 1、7、15 和 30 天分离的。这项工作的主要重点是属于“对雌二醇的反应”、“对促卵泡激素的反应”、“细胞对激素刺激的反应”、“细胞激素代谢过程”和“激素生物合成过程”基因本体论组的基因。这些基因群与 GC 激素代谢和细胞对激素的反应有关。确定了属于这些组的 80 个基因。那些属于多个分析基因本体论组的成员，或表现出独特表达模式的基因，被选择进行进一步分析。选择了所有选定的基因，并详细描述了它们的表达模式。在本文中，描述了两种基因表达模式。第一种模式显示培养后期基因的大量下调，第二种模式显示 IVC 后第 1 天表达上调。本研究主要集中在被发现对类固醇生成最重要的六个基因上：STAR、POR、CYP11A1、ADM、GCLC、IL1B，以及三个在长期体外培养后期高表达的基因：NR2F2、BMP4、COL1A1。本研究的主要目的是选择参与 GC 长期体外培养中激素刺激和激素代谢反应的基因。
