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首页> 外文期刊>Reproductive Biology and Endocrinology >Genes targeted by the estrogen and progesterone receptors in the human endometrial cell lines HEC1A and RL95-2
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Genes targeted by the estrogen and progesterone receptors in the human endometrial cell lines HEC1A and RL95-2

机译:人子宫内膜细胞系HEC1A和RL95-2中雌激素和孕激素受体靶向的基因

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Background When the steroid hormones estrogen and progesterone bind to nuclear receptors, they have transcriptional impact on target genes in the human endometrium. These transcriptional changes have a critical function in preparing the endometrium for embryo implantation. Methods 382 genes were selected, differentially expressed in the receptive endometrium, to study their responsiveness of estrogen and progesterone. The endometrial cell lines HEC1A and RL95-2 were used as experimental models for the non-receptive and receptive endometrium, respectively. Putative targets for activated steroid hormone receptors were investigated by chromatin immunoprecipitation (ChIP) using receptor-specific antibodies. Promoter occupancy of the selected genes by steroid receptors was detected in ChIP-purified DNA by quantitative PCR (qPCR). Expression analysis by reverse transcriptase (RT)-PCR was used to further investigate hormone dependent mRNA expression regulation of a subset of genes. Results ChIP-qPCR analysis demonstrated that each steroid hormone receptor had distinct group of target genes in the endometrial cell lines. After estradiol treatment, expression of estrogen receptor target genes predominated in HEC1A cells (n = 137) compared to RL95-2 cells (n = 35). In contrast, expression of progesterone receptor target genes was higher in RL95-2 cells (n = 83) than in HEC1A cells (n = 7) after progesterone treatment. RT-PCR analysis of 20 genes demonstrated transcriptional changes after estradiol or progesterone treatment of the cell lines. Conclusions Combined results from ChIP-qPCR and RT-PCR analysis showed different patterns of steroid hormone receptor occupancy at target genes, corresponding to activation or suppression of gene expression after hormone treatment of HEC1A and RL95-2 cell lines.
机译:背景技术当类固醇激素雌激素和孕激素与核受体结合时,它们对人子宫内膜的靶基因有转录影响。这些转录改变在准备用于胚胎植入的子宫内膜中具有关键功能。方法选择382个在受体内膜中差异表达的基因,研究其对雌激素和孕激素的反应。子宫内膜细胞系HEC1A和RL95-2分别用作非子宫内膜和子宫内膜的实验模型。使用受体特异性抗体通过染色质免疫沉淀(ChIP)研究了激活的类固醇激素受体的假定靶标。通过定量PCR(qPCR)在ChIP纯化的DNA中检测到类固醇受体对所选基因的启动子占有。通过逆转录酶(RT)-PCR的表达分析被用于进一步研究基因子集的激素依赖性mRNA表达调控。结果ChIP-qPCR分析表明,每个类固醇激素受体在子宫内膜细胞系中都有不同的靶基因组。雌二醇处理后,与RL95-2细胞(n = 35)相比,HEC1A细胞(n = 137)中雌激素受体靶基因的表达占主导地位。相反,经孕酮处理后,RL95-2细胞(n = 83)中的孕酮受体靶基因表达高于HEC1A细胞(n = 7)。对20个基因的RT-PCR分析表明,雌二醇或孕酮处理细胞系后,转录发生了变化。结论ChIP-qPCR和RT-PCR分析的综合结果显示,激素基因对HEC1A和RL95-2细胞系的激活或抑制后,靶基因上甾体激素受体的占据方式不同。

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