From the College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang 310058, China.
the Laboratory of Aquaculture and Artemia Reference Center, Department of Animal Production, Ghent University, B-9000 Ghent, Belgium, and.
J Biol Chem. 2019 Apr 19;294(16):6598-6611. doi: 10.1074/jbc.RA118.005900. Epub 2019 Feb 14.
Cellular quiescence, a reversible state in which growth, proliferation, and other cellular activities are arrested, is important for self-renewal, differentiation, development, regeneration, and stress resistance. However, the physiological mechanisms underlying cellular quiescence remain largely unknown. In the present study, we used embryos of the crustacean in the diapause stage, in which these embryos remain quiescent for prolonged periods, as a model to explore the relationship between cell-membrane potential () and quiescence. We found that is hyperpolarized and that the intracellular chloride concentration is high in diapause embryos, whereas is depolarized and intracellular chloride concentration is reduced in postdiapause embryos and during further embryonic development. We identified and characterized the chloride ion channel protein cystic fibrosis transmembrane conductance regulator (CFTR) of (CFTR) and found that its expression is silenced in quiescent cells of diapause embryos but remains constant in all other embryonic stages. -CFTR knockdown and GlyH-101-mediated chemical inhibition of CFTR produced diapause embryos having a high and intracellular chloride concentration, whereas control embryos released free-swimming nauplius larvae. Transcriptome analysis of embryos at different developmental stages revealed that proliferation, differentiation, and metabolism are suppressed in diapause embryos and restored in postdiapause embryos. Combined with RNA sequencing (RNA-Seq) of GlyH-101-treated MCF-7 breast cancer cells, these analyses revealed that CFTR inhibition down-regulates the Wnt and Aurora Kinase A (AURKA) signaling pathways and up-regulates the p53 signaling pathway. Our findings provide insight into CFTR-mediated regulation of cellular quiescence and in the model.
细胞静止是一种生长、增殖和其他细胞活动停滞的可逆状态，对于自我更新、分化、发育、再生和应激抵抗很重要。然而，细胞静止的生理机制在很大程度上仍然未知。在本研究中，我们使用处于休眠阶段的甲壳动物 的胚胎作为模型，探索细胞膜电位 ( ) 与静止之间的关系。我们发现休眠胚胎中的 处于超极化状态，细胞内氯离子浓度较高，而在休眠后胚胎和进一步的胚胎发育过程中， 去极化，细胞内氯离子浓度降低。我们鉴定并表征了 的氯离子通道蛋白囊性纤维化跨膜电导调节因子 (CFTR) (CFTR)，并发现其在休眠细胞中的表达被沉默，但在所有其他胚胎阶段保持不变。 -CFTR 敲低和 GlyH-101 介导的 CFTR 化学抑制导致休眠胚胎具有高 和细胞内氯离子浓度，而对照 胚胎释放自由游动的无节幼体幼虫。不同发育阶段胚胎的转录组分析表明，增殖、分化和代谢在休眠胚胎中受到抑制，而在休眠后胚胎中得到恢复。结合 GlyH-101 处理的 MCF-7 乳腺癌细胞的 RNA 测序 (RNA-Seq) 分析，这些分析表明 CFTR 抑制下调 Wnt 和 Aurora Kinase A (AURKA) 信号通路，并上调 p53 信号通路。我们的研究结果为 CFTR 介导的 模型中细胞静止和 的调控提供了新的见解。
