摘要
TG interaction factor 1 (TGIF1) plays a major role in transcriptional inhibition and suppression of TGF-β signaling, but its functional roles in granulosa cells (GCs) have not been elucidated; in particular, there is no information about the yak (Bos grunniens) TGIF1 gene. Therefore, the objectives of this study were to clone yak TGIF1 and investigate TGIF1 functions in yak GCs. RT‒PCR results showed that the coding region of yak TGIF1 is 759 bp and encodes 252 amino acids. Its nucleotide sequence showed 85.24–99.74% similarity to mouse, human, pig, goat and cattle homologous genes. To explore the functional roles of TGIF1, we studied proliferation, apoptosis, cell cycle progression, steroidogenesis and the expression levels of related genes in yak GCs transfected with small interfering RNA specific to TGIF1. The results showed that TGIF1 knockdown promoted proliferation and cell cycle progression and inhibited apoptosis and estradiol (E2) and progesterone (P4) production in cultured yak GCs. Conversely, TGIF1 overexpression inhibited proliferation and cell cycle progression and stimulated apoptosis and E2 and P4 production. In addition, these functional changes in yak GCs were observed parallel to the expression changes in genes involved in the cell cycle (PCNA, CDK2, CCND1, CCNE1, CDK4 and P53), apoptosis (BCL2, BAX and CASPASE3), and steroidogenesis (CYP11A1, 3β-HSD and StAR). In conclusion, TGIF1 was relatively conserved in the course of animal evolution. TGIF1 inhibited GC viability and stimulated apoptosis and the secretion of E2 and P4 by yak GCs. Our results will help to reveal the mechanism underlying yak follicular development and improve the reproductive efficiency of female yaks.
摘要译文
TG相互作用因子1（TGIF1）在转录抑制和抑制TGF-β信号传导中起主要作用，但尚未阐明其在颗粒细胞（GC）中的功能作用。特别是，没有关于牛（BOS grunniens）TGIF1基因的信息。因此，这项研究的目标是克隆Yak TGIF1并研究YAK GC中的TGIF1功能。RT = PCR的结果表明，Yak TGIF1的编码区为759 bp，编码252个氨基酸。它的核苷酸序列显示出与小鼠，人，猪，山羊和牛同源基因的85.24–99.74％的相似性。为了探索TGIF1的功能作用，我们研究了在Yak GC中使用特定于TGIF1的小干扰RNA转染的YAK GC中相关基因的增殖，细胞周期进展，类固醇生成和相关基因的表达水平。结果表明，TGIF1敲低促进了培养的YAK GC中的凋亡和雌二醇（E 2）和孕酮（P 4）产生的细胞周期进程，并抑制了凋亡和雌二醇（E 2）。相反，TGIF1的过表达抑制了增殖和细胞周期进程，并刺激了凋亡以及E 2和P 4的产生。此外，观察到与细胞周期中涉及的基因的表达变化平行观察到的这些功能变化（PCNA，CDK2，CCND1，CCND1，CCNE1，CDK4和P53），凋亡（Bcl2，Bax和caspase3）和类固醇生成（CYP11A11A11A1），3β-HSD和Star）。总之，TGIF1在动物进化过程中相对保守。TGIF1抑制了GC的生存力和刺激的凋亡以及E 2和P 4的YAK GC分泌。我们的结果将有助于揭示牛卵泡发育的基础机制，并提高雌性牛的生殖效率。
Xiang-dong Zi [a]. Molecular characterization and effects of the TGIF1 gene on proliferation and steroidogenesis in yak (Bos grunniens) granulosa cells[J]. Theriogenology, 2023,211: 224-231