Establishment of a Human Nonluteinized Granulosa Cell Line that Transitions from the Gonadotropin-Independent to the Gonadotropin-Dependent Status
Establishment of a Human Nonluteinized Granulosa Cell Line that Transitions from the Gonadotropin-Independent to the Gonadotropin-Dependent Status
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DOI：
10.1210/en.2011-1810
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发表时间：
2012-06-01
期刊：
影响因子：
4.8
通讯作者：
Kikkawa, Fumitaka
中科院分区：
文献类型：
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作者：
Bayasula;Iwase, Akira;Kikkawa, Fumitaka
卵巢是一个复杂的内分泌器官，负责类固醇和卵泡的生成。卵泡包括卵母细胞和两种主要的类固醇生成细胞类型，颗粒细胞和卵泡膜细胞。永生化的人颗粒细胞是研究类固醇激素和卵泡发生机制的基础。我们从一名35岁的女性获得颗粒细胞，并通过慢病毒介导的几种基因转移使其永生化，以建立人非黄素化颗粒细胞系（HGrC 1）。我们随后表征了HGrC 1并研究了其类固醇生成性能。HGrC 1表达与类固醇生成相关的酶，如类固醇生成急性调节蛋白、CYP 11 A、芳香化酶和促性腺激素受体。FSH刺激增加芳香化酶的mRNA水平，从而诱导雄烯二酮芳构化为雌二醇。激活素A增加FSH受体的mRNA水平，其与FSH刺激协同上调。HGrC 1还表达一系列属于TGF-β超家族的配体和受体。蛋白质印迹分析表明，骨形态发生蛋白（BMP）-4，BMP-6和BMP-7磷酸化小母亲对decapentaplegic（Smad）1/5/8，而生长分化因子-9磷酸化Smad 2/3。BMP-15和抗苗勒管激素磷酸化Smad 1/5/8，同时也弱磷酸化Smad 2/3。提示HGrC 1可能具有早期卵泡颗粒细胞的特征。HGrC 1也可能能够显示从促性腺激素非依赖性状态到促性腺激素依赖性状态的生长转变。（内分泌学153：2851-2860，2012）
The ovary is a complex endocrine organ responsible for steroidogenesis and folliculogenesis. Follicles consist of oocytes and two primary steroidogenic cell types, the granulosa cells, and the theca cells. Immortalized human granulosa cells are essential for researching the mechanism of steroidogenesis and folliculogenesis. We obtained granulosa cells from a 35-yr-old female and immortalized them by lentivirus-mediated transfer of several genes so as to establish a human nonluteinized granulosa cell line (HGrC1). We subsequently characterized HGrC1 and investigated its steroidogenic performance. HGrC1 expressed enzymes related to steroidogenesis, such as steroidogenic acute regulatory protein, CYP11A, aromatase, and gonadotropin receptors. Stimulation with FSH increased the mRNA levels of aromatase, which consequently induced the aromatization of androstenedione to estradiol. Activin A increased the mRNA levels of the FSH receptor, which were synergistically up-regulated with FSH stimulation. HGrC1 also expressed a series of ligands and receptors belonging to the TGF-beta superfamily. A Western blot analysis showed that bone morphogenetic protein (BMP)-4, BMP-6, and BMP-7 phosphorylated small mother against decapentaplegic (Smad) 1/5/8, whereas growth differentiation factor-9 phosphorylated Smad2/3. BMP-15 and anti-Mullerian hormone phosphorylated Smad1/5/8 while also weakly phosphorylating Smad2/3. These results indicate that HGrC1 may possess the characteristics of granulosa cells belonging to follicles in the early stage. HGrC1 might also be capable of displaying the growth transition from a gonadotropin-independent status to gonadotropin-dependent one. (Endocrinology 153: 2851-2860, 2012)