Tremblay Patricia G, Sirard Marc-André
Biol Reprod. 2017 Apr 1;96(4):855-865. doi: 10.1093/biolre/iox024.
The developmental competence of an oocyte is its capacity to resume maturation, undergo successful fertilization, and reach the blastocyst stage. This competence is acquired through interaction with somatic cells of the follicle. Cumulus and granulosa cells support oocyte development, while the oocyte influences follicular cell growth and differentiation. Studies suggest that follicle-stimulating hormone and luteinizing hormone play an essential role in oocyte competence acquisition through signaling initiated by protein kinases A and C (PKA and PKC) in granulosa cells. Using a microarray and RT-qPCR, the transcriptome of human granulosa-like tumor cells (KGN) treated for 24 h with forskolin (FSK) or phorbol 12-myristate 13-acetate (PMA) was analyzed to determine the effects of PKA and PKC stimulation on gene expression. Protein-kinase-driven signaling appeared to involve five major upstream regulators, namely epidermal growth factor (EGF), transforming growth factor beta 1 (TGFβ1), vascular endothelial growth factor (VEGF), basic fibroblast growth factor (FGF2), and hepatocyte growth factor (HGF). Gene associations with seven major ovarian functions were identified: Prostaglandin- endoperoxide synthase 2 (PTGS2), interleukin 8 (IL8), and interleukin 6 (IL6) with inflammation; Steroidogenic acute regulatory protein (STAR), cytochrome P450scc (CYP11A1), and cytochrome P450 family 19 subfamily A member 1 (CYP19A1) with steroidogenesis; Vascular endothelial growth factor C (VEGFC), Vascular endothelial growth factor A (VEGFA), and C-X-C chemokine receptor type 4 (CXCR4) with angiogenesis; Amphiregulin (AREG), epidermal growth factor receptor (EGFR), and sprouty RTK signaling antagonist 2 (SPRY2) with differentiation, BCL2 associated X (BAX), BCL2 like 12 (BCL2L12), and caspase 1(CASP1) with apoptosis, Cyclin D1 (CCND1), cyclin B1 (CCNB1), and cyclin B2 (CCNB2) with division; and Matrix metalloproteinase-1 (MMP1), Matrix metallopeptidase 9 (MMP9), and TIMP metallopeptidase inhibitor 1 (TIMP1) with ovulation. Overall, these results indicate that signaling via both PKA and PKC potentiates gene regulation of functions such as inflammation and apoptosis, while functions such as differentiation, ovulation and angiogenesis are partial to one kinase or the other. These results improve understanding of the pathways underlying the most important changes that occur in the follicle prior to ovulation.
卵母细胞的发育能力是其恢复成熟、成功受精并达到囊胚阶段的能力。这种能力是通过与卵泡的体细胞相互作用而获得的。卵丘细胞和颗粒细胞支持卵母细胞发育，而卵母细胞则影响卵泡细胞的生长和分化。研究表明，促卵泡激素和促黄体生成素通过颗粒细胞中蛋白激酶A和C（PKA和PKC）启动的信号传导，在卵母细胞能力获得中起重要作用。使用微阵列和RT-qPCR分析了用福斯可林（FSK）或佛波酯12-肉豆蔻酸酯13-乙酸酯（PMA）处理24小时的人颗粒样肿瘤细胞（KGN）的转录组，以确定PKA和PKC刺激对基因表达的影响。蛋白激酶驱动的信号传导似乎涉及五个主要的上游调节因子，即表皮生长因子（EGF）、转化生长因子β1（TGFβ1）、血管内皮生长因子（VEGF）、碱性成纤维细胞生长因子（FGF2）和肝细胞生长因子（HGF）。确定了与七种主要卵巢功能相关的基因：前列腺素内过氧化物合酶2（PTGS2）、白细胞介素8（IL8）和白细胞介素6（IL6）与炎症相关；类固醇生成急性调节蛋白（STAR）、细胞色素P450侧链裂解酶（CYP11A1）和细胞色素P450家族19亚家族A成员1（CYP19A1）与类固醇生成相关；血管内皮生长因子C（VEGFC）、血管内皮生长因子A（VEGFA）和C-X-C趋化因子受体4（CXCR4）与血管生成相关；双调蛋白（AREG）、表皮生长因子受体（EGFR）和Sprouty RTK信号拮抗剂2（SPRY2）与分化相关，BCL2相关X蛋白（BAX）、BCL2样蛋白12（BCL2L12）和半胱天冬酶1（CASP1）与凋亡相关，细胞周期蛋白D1（CCND1）、细胞周期蛋白B1（CCNB1）和细胞周期蛋白B2（CCNB2）与分裂相关；基质金属蛋白酶-1（MMP1）、基质金属蛋白酶9（MMP9）和金属蛋白酶组织抑制剂1（TIMP1）与排卵相关。总体而言，这些结果表明，通过PKA和PKC的信号传导增强了炎症和凋亡等功能的基因调控，而分化、排卵和血管生成等功能则部分依赖于一种激酶或另一种激酶。这些结果有助于更好地理解排卵前卵泡中发生的最重要变化的潜在途径。
