Department of Clinical Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling 712100, Shaanxi, China.
Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Northwest A&F University, Yangling 712100, Shaanxi, China.
Int J Mol Sci. 2018 Nov 8;19(11):3517. doi: 10.3390/ijms19113517.
CREBZF, a multifunction transcriptional regulator, participates in the regulation of numerous cellular functions. The aims of the present study were to detect the localization of CREBZF expression in the ovary and explore the role of CREBZF and related mechanisms in the apoptosis of ovarian granulosa cells. We found by immunohistochemistry that CREBZF was mainly located in granulosa cells and oocytes during the estrous cycle. Western blot analysis showed that SMILE was the main isoform of CREBZF in the ovary. The relationship between apoptosis and CREBZF was assessed via CREBZF overexpression and knockdown. Flow cytometry analysis showed that CREBZF induced cell apoptosis in granulosa cells. Western bolt analysis showed that overexpression of CREBZF upregulated BAX and cleaved Caspase-3, while it downregulated BCL-2. Furthermore, overexpression of CREBZF inhibited the ERK1/2 and mTOR signaling pathways through the phosphorylation of intracellular-regulated kinases 1/2 (ERK1/2) and p70 S6 kinase (S6K1). Moreover, we found that CREBZF also activated autophagy by increasing LC3-II. In summary, these results suggest that CREBZF might play a proapoptotic role in cell apoptosis in granulosa cells, possibly by regulating the ERK1/2 and mTOR signaling pathways.
CREBZF 是一种多功能转录调节因子，参与众多细胞功能的调节。本研究旨在检测 CREBZF 在卵巢中的表达定位，并探讨 CREBZF 及其相关机制在卵巢颗粒细胞凋亡中的作用。通过免疫组织化学染色发现，CREBZF 在发情周期中主要定位于颗粒细胞和卵母细胞中。Western blot 分析显示，SMILE 是卵巢中 CREBZF 的主要同工型。通过 CREBZF 的过表达和敲低评估凋亡与 CREBZF 之间的关系。流式细胞术分析显示 CREBZF 诱导颗粒细胞凋亡。Western blot 分析显示，CREBZF 的过表达上调了 BAX 和 cleaved Caspase-3，而下调了 BCL-2。此外，CREBZF 的过表达通过细胞内调节激酶 1/2（ERK1/2）和 p70 S6 激酶（S6K1）的磷酸化抑制了 ERK1/2 和 mTOR 信号通路。此外，我们发现 CREBZF 通过增加 LC3-II 还激活了自噬。总之，这些结果表明，CREBZF 可能在颗粒细胞凋亡中发挥促凋亡作用，可能通过调节 ERK1/2 和 mTOR 信号通路。
