EZH2 Gene Knockdown Inhibits Sheep Pituitary Cell Proliferation via Downregulating the AKT/ERK Signaling Pathway

• Published in: International journal of molecular sciences 2023-06, Vol.24 (13), p.10656
• Main Authors: Cai, Yu, Chen, Peiyong, Xu, Hui, Li, Shanglai, Zhao, Bingru, Fan, Yixuan, Wang, Feng, Zhang, Yanli
• Format: Article
• Language: English
• Subjects: AKT protein; Animals; Apoptosis; BAX protein; Cell cycle; Cell growth; Cell proliferation; Cell Proliferation - genetics; Epigenetics; EZH2; EZH2 gene; Fecundity; Fertility; Fetuses; Flow cytometry; Follicle Stimulating Hormone, beta Subunit - genetics; Follicle-stimulating hormone; Gametogenesis; Gene expression; Gene Knockdown Techniques; gonadotropin secretion; Gonadotropins; Histone methyltransferase; Hu sheep; Hypothalamus; Mammals - genetics; Molecular modelling; Morphology; Ovarian cancer; Ovaries; Ovulation; Phosphorylation; Physiology; Pituitary; Pituitary (anterior); pituitary cells; proliferation; Proteins; Proto-Oncogene Proteins c-akt - genetics; Relative abundance; Sheep; Sheep - genetics; Signal transduction; Signal Transduction - physiology; Stem cells; Steroidogenesis; TOR protein; Tumors
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms241310656

Pituitary gonadotropins perform essential functions in mammalian reproduction by stimulating gametogenesis and steroidogenesis in the ovaries and testicles. EZH2 is a histone methyltransferase that inhibits proliferation and aggravates apoptosis in stem cells subjected to pathological stimuli. However, the expression and molecular mechanisms of EZH2 in pituitary cells in vitro have not been extensively studied. In this study, the relative abundances of EZH2 mRNA ( < 0.01) and protein ( < 0.05) expression were larger in the pituitary cells of Hu sheep with relatively greater fecundity (GF) compared to those with lesser fecundity (LF). Loss-of-function examinations demonstrated that gene knockdown led to an earlier induction of apoptosis in sheep pituitary cells (PCs). The relative abundance of , , and was increased ( < 0.01), while 's abundance was less decreased ( < 0.01) in PCs where there was gene knockdown. Additionally, cell proliferation ( < 0.01) and viability ( < 0.01) were decreased in EZH2-knockdown sheep PCs, and the cell cycle was blocked compared to a negative control (NC). Notably, gene knockdown led to reduced abundances of gonadotropin subunit gene transcripts (FSHβ, < 0.05) and reduced FSH release ( < 0.01) from PCs. gene knockdown led to reduced phosphorylation of AKT, ERK, and mTOR ( < 0.01). The results suggest that EZH2 regulates pituitary cell proliferation, apoptosis, and FSH secretion through modulation of the AKT/ERK signaling pathway, providing a foundation for further study of pituitary cell functions.