Regulation of amphiregulin, EGFR‐like factor expression by hCG in cultured human granulosa cells
Regulation of amphiregulin, EGFR‐like factor expression by hCG in cultured human granulosa cells
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DOI：
10.1080/00016340701314959
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发表时间：
2007-06
影响因子：
4.3
通讯作者：
Hiroaki Negishi;Chiaki Ikeda;Yoko Nagai;Akiko Satoh;Yoko Kumasako;Satoru Makinoda;Takafumi Ustunomiya
中科院分区：
文献类型：
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作者：
Hiroaki Negishi;Chiaki Ikeda;Yoko Nagai;Akiko Satoh;Yoko Kumasako;Satoru Makinoda;Takafumi Ustunomiya
背景。长期以来，人们一直怀疑表皮生长因子（EGF）受体及其一些假定的配体可能在卵巢功能中起重要作用。双调节蛋白（AR）是一种具有EGF样基序的生长因子，但其在卵巢信号传导中的潜在作用尚不清楚。研究了AR基因在人未成熟卵泡颗粒细胞中的表达及其功能作用。方法。从月经期较早的未成熟卵泡中提取颗粒细胞，分别给予或不给予200 mIU/ml FSH刺激和1 IU/ml hCG刺激进行培养。应用RT - PCR半定量测定AR和促黄体生成素替代（LHR） mRNA水平。测定培养基中黄体酮(P)浓度。结果。FSH刺激48 h后LHR mRNA表达，AR mRNA不表达。AR mRNA在hCG刺激后1 h表达，并在6 h内表达强度增加。AR增加P的生物合成呈剂量依赖性。在没有FSH和hCG刺激的情况下，forskolin刺激可使AR mRNA在LHR mRNA表达前升高。当FSH培养15小时，然后增加hCG刺激剂量6小时时，AR mRNA水平随hCG浓度增加而增加，最高可达1,000 mIU/ml。结论。FSH刺激后出现LHR基因表达是AR基因在体内表达的必要条件，体外不表达LHR基因的forskolin诱导AR基因表达。AR在一定程度上刺激了P的生物合成，提示AR对颗粒细胞的分化作用。AR可能是排卵前LH信号的中介。
Background. It has long been suspected that the epidermal growth factor (EGF) receptor and some of its putative ligands may play an important role in ovarian function. Amphiregulin (AR) is the growth factor with an EGF‐like motif, but its potential role in signalling in the ovary is still obscure. AR gene expression and its functional effect were evaluated in human granulosa cells from immature follicles. Method. Granulosa cells from immature follicles with early menstrual phase were cultured with or without 200 mIU/ml of FSH stimulation, following with or without 1 IU/ml of hCG. mRNA levels of AR and luteinising hormone replacement (LHR) were semi‐quantified using RT‐PCR. Progesterone (P) concentration in the medium was assayed. Results. LHR mRNA was expressed 48 h after FSH stimulation without AR mRNA expression. AR mRNA was expressed 1 h after hCG stimulation, and increased the intensity in 6 h. P biosynthesis was increased by AR in a dose‐dependent manner. AR mRNA was elevated by forskolin stimulation without FSH and hCG stimulation before LHR mRNA expression. When cultured with FSH for 15 h, followed by increasing doses of hCG stimulation for 6 h, the AR mRNA levels increased according to hCG concentration up to 1,000 mIU/ml. Conclusion. Occurrence of LHR gene expression following FSH stimulation was necessary for the AR gene expression in vivo, and the AR gene was induced by forskolin without LHR gene expression in vitro. P biosynthesis was stimulated, to some extent, by AR. This result suggests the differentiation effect of AR on granulosa cells. AR might be a mediator of LH signals before ovulation.