Chung Daesuk, Gao Fei, Jegga Anil G, Das Sanjoy K
Division of Reproductive Sciences, Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, Cincinnati, OH 45229, USA; The Perinatal Institute, Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, Cincinnati, OH 45229, USA.
Biomedical Informatics, Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, Cincinnati, OH 45229, USA.
Mol Cell Endocrinol. 2015 Jan 15;400:48-60. doi: 10.1016/j.mce.2014.11.002. Epub 2014 Nov 7.
To define endometrial stromal-derived paracrine mediators that participate in estradiol-17β (E2)-induced epithelial proliferation, microarray analysis of gene expression was carried out in mouse uterine epithelial-stromal co-culture systems under the condition of E2 or vehicle (control). Our results demonstrated gene alteration by E2: in epithelial cells, we found up-regulation of 119 genes and down-regulation of 28 genes, while in stroma cells we found up-regulation of 144 genes and down-regulation of 184 genes. A functional enrichment analysis of the upregulated epithelial genes implicated them for proliferation, while upregulated stromal genes were associated with extracellular functions. Quantitative RT-PCR and in situ hybridization results confirmed differential gene expression in both cell cultures and ovariectomized uteri after the above treatments. Based on our identification of stromal secretory factors, we found evidence that suppression by siRNA specifically for Bmp8a and/or Fgf10 in the stromal layer caused significant inhibition of proliferation by E2 in the co-culture system, suggesting Bmp8a and Fgf10 act as paracrine mediators during E2-dependent control of uterine proliferation. The localization of receptors and receptor activation signaling in epithelial cells in both the co-culture system and uteri was consistent with their involvement in ligand-receptor signaling. Interestingly, loss of Bmp8a or Fgf10 also caused abrogation of E2-regulated epithelial receptor signaling in co-culture systems, suggesting that stroma-derived Fgf10 and Bmp8a are responsible for epithelial communication. Overall, stromal Fgf10 and Bmp8a serve as potential paracrine factors for E2-dependent regulation of epithelial proliferation in the uterus.
为了确定参与雌二醇-17β（E2）诱导的上皮细胞增殖的子宫内膜基质衍生旁分泌介质，在E2或溶剂（对照）条件下，对小鼠子宫上皮-基质共培养系统进行了基因表达微阵列分析。我们的结果显示了E2引起的基因改变：在上皮细胞中，我们发现119个基因上调，28个基因下调，而在基质细胞中，我们发现144个基因上调，184个基因下调。对上调的上皮基因进行的功能富集分析表明它们与增殖有关，而上调的基质基因与细胞外功能有关。定量RT-PCR和原位杂交结果证实了上述处理后两种细胞培养物和去卵巢子宫中的差异基因表达。基于我们对基质分泌因子的鉴定，我们发现证据表明，在基质层中特异性针对Bmp8a和/或Fgf10的siRNA抑制会导致共培养系统中E2对增殖的显著抑制，这表明Bmp8a和Fgf10在E2依赖性子宫增殖控制过程中作为旁分泌介质发挥作用。共培养系统和子宫中上皮细胞中受体的定位以及受体激活信号与它们参与配体-受体信号传导一致。有趣的是，Bmp8a或Fgf10的缺失也导致共培养系统中E2调节的上皮受体信号传导的废除，这表明基质衍生的Fgf10和Bmp8a负责上皮细胞间通讯。总体而言，基质Fgf10和Bmp8a作为E2依赖性调节子宫上皮增殖的潜在旁分泌因子。
