Herndon Maria K, Law Nathan C, Donaubauer Elyse M, Kyriss Brandon, Hunzicker-Dunn Mary
School of Molecular Biosciences and the Center for Reproductive Biology, Washington State University, Pullman WA 99163, USA.
Mol Cell Endocrinol. 2016 Oct 15;434:116-26. doi: 10.1016/j.mce.2016.06.020. Epub 2016 Jun 17.
FSH promotes maturation of ovarian follicles. One pathway activated by FSH in granulosa cells (GCs) is phosphatidylinositol-3 kinase/AKT. The AKT target FOXO1 is reported to function primarily as a repressor of FSH genes, including Ccnd2 and Inha. Based on its broad functions in other tissues, we hypothesized that FOXO1 may regulate many more GC genes. We transduced GCs with empty adenovirus or constitutively active FOXO1 followed by treatment with FSH for 24 h, and conducted RNA deep sequencing. Results show that FSH regulates 3772 genes ≥2.0-fold; 60% of these genes are activated or repressed by FOXO1. Pathway Studio Analysis revealed enrichment of genes repressed by FOXO1 in metabolism, signaling, transport, development, and activated by FOXO1 in signaling, cytoskeletal functions, and apoptosis. Gene regulation was verified by q-PCR (eight genes) and ChIP analysis (two genes). We conclude that FOXO1 regulates the majority of FSH target genes in GCs.
促卵泡生成素（FSH）促进卵巢卵泡成熟。FSH在颗粒细胞（GCs）中激活的一条信号通路是磷脂酰肌醇-3激酶/蛋白激酶B（AKT）。据报道，AKT的靶标叉头框蛋白O1（FOXO1）主要作为包括细胞周期蛋白D2（Ccnd2）和抑制素α（Inha）在内的FSH基因的阻遏物发挥作用。基于其在其他组织中的广泛功能，我们推测FOXO1可能调控更多的GC基因。我们用空腺病毒或组成型激活的FOXO1转导GCs，然后用FSH处理24小时，并进行RNA深度测序。结果显示，FSH调控3772个基因，其表达变化≥2.0倍；这些基因中有60%被FOXO1激活或抑制。通路工作室分析显示，FOXO1抑制的基因在代谢、信号传导、转运、发育等方面富集，而FOXO1激活的基因在信号传导、细胞骨架功能和细胞凋亡方面富集。通过定量聚合酶链反应（q-PCR，检测8个基因）和染色质免疫沉淀分析（ChIP，检测2个基因）验证了基因调控情况。我们得出结论，FOXO1调控GCs中大多数FSH靶基因。
