Wu Fangfang, Liu Zhihui, Huang Jing, Gao Yuan, Yang Lan, Du Fuliang
Jiangsu Key Laboratory for Molecular and Medical Biotechnology, College of Life Sciences, Nanjing Normal University, #1 Wenyuan Rd, Nanjing, 210046, China.
Chengdu Institute of Biological Products Co., Ltd., 379, 3rd Section, Jinhua Road, Jinjiang District, Chengdu, 610023, Sichuan Province, China.
Sci Rep. 2025 Jul 26;15(1):27204. doi: 10.1038/s41598-025-12427-3.
Polycomb repressive complexes (PRC) and nucleosome remodeling and deacetylase (NuRD) complex are crucial for regulating the expression of pluripotent and developmental genes and maintaining the characteristics of mouse embryonic stem cells (mESCs). However, the interplay between the Polycomb and NuRD complexes in mESCs, particularly under protein kinase C (PKC) inhibition, remains to be elucidated. We knocked down Polycomb complexes components Ezh2, Ring1b, and Cbx7 via short hairpin RNA interference and observed significant reductions in most NuRD complex components, especially Mbd3, Mta1, Rbbp4, and Rbbp7. Similarly, Ezh2 overexpression increased the levels of these major NuRD complex components. Further, Mbd3 knockdown significantly reduced the expression of PRC1 major components Ring1b, Rybp, and Cbx7 and PRC2 major components Ezh2, Suz12, and Eed, but its overexpression had no significant effect on their levels. These results indicate that PKC inhibition provides a suitable environment for the expression of PRC components. Altogether, our study demonstrates that mESCs exhibit mutual gene regulation of Polycomb and NuRD complexes under PKC inhibition that maintains pluripotency and self-renewal abilities and regulates the plasticity of mESCs to balance between pluripotency and cell fate determination.
多梳抑制复合物(PRC)和核小体重塑与去乙酰化酶(NuRD)复合物对于调节多能性和发育相关基因的表达以及维持小鼠胚胎干细胞(mESCs)的特性至关重要。然而,PRC和NuRD复合物在mESCs中的相互作用,特别是在蛋白激酶C(PKC)抑制作用下的相互作用,仍有待阐明。我们通过短发夹RNA干扰敲低了多梳复合物的组分Ezh2、Ring1b和Cbx7,并观察到大多数NuRD复合物组分显著减少,尤其是Mbd3、Mta1、Rbbp4和Rbbp7。同样,Ezh2的过表达增加了这些主要NuRD复合物组分的水平。此外,Mbd3的敲低显著降低了PRC1主要组分Ring1b、Rybp和Cbx7以及PRC2主要组分Ezh2、Suz12和Eed的表达,但其过表达对它们的水平没有显著影响。这些结果表明,PKC抑制为PRC组分的表达提供了适宜的环境。总之,我们的研究表明,mESCs在PKC抑制下表现出PRC和NuRD复合物之间的相互基因调控,这种调控维持了多能性和自我更新能力,并调节了mESCs在多能性和细胞命运决定之间平衡的可塑性。