Fibroblast growth factor 9 (FGF9) regulation of cyclin D1 and cyclin-dependent kinase-4 in ovarian granulosa and theca cells of cattle.
Fibroblast growth factor 9 (FGF9) regulation of cyclin D1 and cyclin-dependent kinase-4 in ovarian granulosa and theca cells of cattle.
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DOI：
10.1016/j.mce.2016.11.002
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发表时间：
2017-01-15
影响因子：
4.1
通讯作者：
Spicer, L. J.
中科院分区：
文献类型：
--
作者：
Totty, M. L.;Morrell, B. C.;Spicer, L. J.
关键词：
为了确定成纤维细胞生长因子9（FGF 9）改变颗粒（GC）和卵泡膜（TC）细胞增殖的机制，评价了调节细胞周期G1期进展的细胞周期蛋白，细胞周期蛋白D1（CCND 1）和细胞周期蛋白依赖性激酶4（CDK 4; CCND 1的催化伴侣）。从当地屠宰场获得卵巢，从小（1-5 mm）和大（8-22 mm）卵泡收获GC，并且从大卵泡收获TC。将GC和TC接种在含有10%胎牛血清的培养基中，然后在无血清培养基中进行各种处理。用30 ng/mL的FGF 9或IGF 1处理显著增加GC数量，并且当组合时，协同作用以进一步增加GC数量三倍。实时荧光定量PCR检测TC和GC中CCND 1和CDK 4 mRNA的表达。单独和与IGF 1组合，FGF 9显著增加GC和TC中的CCND 1 mRNA表达。Western blotting结果显示，TGF-9处理6 h和12 h后，TC中CCND 1蛋白表达增加，而CDK 4蛋白表达无明显变化。丝裂原活化蛋白激酶（MAPK）/细胞外信号调节激酶（ERK）通路抑制剂U 0126可显著降低FGF 9诱导的CCND 1 mRNA表达至基础水平。我们首次表明，CCND 1 mRNA的表达增加了FGF 9在牛TC和GC，和FGF 9可能使用MAPK途径诱导CCND 1 mRNA的生产在牛TC。
To determine the mechanism by which fibroblast growth factor 9 (FGF9) alters granulosa (GC) and theca (TC) cell proliferation, cell cycle proteins that regulate progression through G1 phase of the cell cycle, cyclin D1 (CCND1) and cyclin-dependent kinase-4 (CDK4; CCND1's catalytic partner), were evaluated. Ovaries were obtained from a local abattoir, GC were harvested from small (1-5 mm) and large (8-22 mm) follicles, and TC were harvested from large follicles. GC and TC were plated in medium containing 10% fetal calf serum followed by various treatments in serum-free medium. Treatment with 30 ng/mL of either FGF9 or IGF1 significantly increased GC numbers and when combined, synergized to further increase GC numbers by threefold. Abundance of CCND1 and CDK4 mRNA in TC and GC were quantified via real-time PCR. Alone and in combination with IGF1, FGF9 significantly increased CCND1 mRNA expression in both GC and TC. Western blotting revealed that CCND1 protein levels were increased by FGF9 in TC after 6 h and 12 h of treatment, but CDK4 protein was not affected. A mitogen-activated protein kinase (MAPK)/ extracellular signal-regulated kinase (ERK) pathway inhibitor, U0126, significantly reduced FGF9-induced CCND1 mRNA expression to basal levels. For the first time we show that CCND1 mRNA expression is increased by FGF9 in bovine TC and GC, and that FGF9 likely uses the MAPK pathway to induce CCND1 mRNA production in bovine TC.
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影响因子：
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