Yuan Xiaolong, Zhou Xiaofeng, He Yingting, Zhong Yuyi, Zhang Ailing, Zhang Zhe, Zhang Hao, Li Jiaqi
Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, National Engineering Research Centre for Breeding Swine Industry, College of Animal Science, South China Agricultural University, Guangzhou 510642, China.
College of Biological and Food Engineering, Guangdong University of Education, Guangzhou 510303, China.
Genes (Basel). 2018 Jun 13;9(6):295. doi: 10.3390/genes9060295.
Previous studies suggest that signal transducer and activator of transcription 3 and CCAAT/enhancer binding protein beta play an essential role in ovarian granulosa cells (GCs) for mammalian follicular development. Several C/EBPβ putative binding sites were previously predicted on the promoter in mammals. However, the molecular regulation of on and their effects on cell proliferation and apoptosis remain virtually unexplored in GCs. Using porcine GCs as a model, the 5′-deletion, luciferase report assay, mutation, chromatin immunoprecipitation, Annexin-V/PI staining and EdU assays were applied to investigate the molecular mechanism for regulating the expression of and their effects on the cell proliferation and apoptosis ability. We found that over and interfering with the expression of significantly increased and decreased the messenger RNA (mRNA) and protein levels of , respectively. The dual luciferase reporter assay showed that C/EBPβ directly bound at −1397/−1387 of to positively regulate the mRNA and protein expressions of . Both and were observed to inhibit cell apoptosis and promote cell proliferation. Furthermore, might enhance the antiapoptotic and pro-proliferative effects of . These results would be of great insight in further exploring the molecular mechanism of and on the function of GCs and the development of ovarian follicles in mammals.
先前的研究表明，信号转导子和转录激活子3以及CCAAT/增强子结合蛋白β在哺乳动物卵泡发育的卵巢颗粒细胞（GCs）中起重要作用。先前在哺乳动物的启动子上预测到了几个C/EBPβ假定结合位点。然而，在GCs中，关于它们对细胞增殖和凋亡的分子调控及其影响实际上仍未得到探索。以猪GCs为模型，应用5′-缺失、荧光素酶报告基因检测、突变、染色质免疫沉淀、Annexin-V/PI染色和EdU检测来研究调控的分子机制及其对细胞增殖和凋亡能力的影响。我们发现，过表达和干扰的表达分别显著增加和降低了的信使核糖核酸（mRNA）和蛋白质水平。双荧光素酶报告基因检测表明，C/EBPβ直接结合在的−1397/−1387处，以正向调节的mRNA和蛋白质表达。和均被观察到抑制细胞凋亡并促进细胞增殖。此外，可能增强的抗凋亡和促增殖作用。这些结果对于进一步探索和对GCs功能及哺乳动物卵泡发育的分子机制具有重要的启示意义。
