Department of Brain and Cognitive Science, DGIST, Daegu, South Korea.
Core Protein Resources Center, DGIST, Daegu, South Korea.
Cell Biochem Funct. 2021 Jan;39(1):67-76. doi: 10.1002/cbf.3554. Epub 2020 Jun 11.
Mouse embryonic stem cells (mESCs) are pluripotent cells that possess the ability to self-renew and differentiate into three germ layers. Owing to these characteristics, mESCs act as important models for stem cell research and are being used in many clinical applications. Among the many cathepsins, cathepsin A (Ctsa), a serine protease, affects the function and properties of stem cells. However, studies on the role of Ctsa in stem cells are limited. Here, we observed a significant increase in Ctsa expression during mESC differentiation at protein levels. Furthermore, we established Ctsa knockdown mESCs. Ctsa knockdown led to Erk1/2 phosphorylation, which in turn inhibited the pluripotency of mESCs and induced G2/M cell cycle arrest to inhibit mESC proliferation. The knockdown also induced abnormal differentiation in mESCs and aberrant expression of differentiation markers. Furthermore, we identified inhibition of teratoma formation in nude mice. Our results suggested that Ctsa affects mESC pluripotency, proliferation, cell cycle and differentiation, and highlighted the potential of Ctsa to act as a core factor that can regulate various mESC properties. SIGNIFICANCE OF THE STUDY: Our results indicate that cathepsin A (Ctsa) affects the properties of mESCs. Inhibition of Ctsa resulted in a decrease in the pluripotency of mouse embryonic stem cells (mESCs). Further, Ctsa suppression resulted in decreased proliferation via cell cycle arrest. Moreover, Ctsa inhibition reduced differentiation abilities and formation of teratoma in mESCs. Our results demonstrated that Ctsa is an important factor controlling mESC abilities.
小鼠胚胎干细胞（mESCs）是多能细胞，具有自我更新和分化为三个胚层的能力。由于这些特性，mESCs 作为干细胞研究的重要模型，正在许多临床应用中得到应用。在许多组织蛋白酶中，组织蛋白酶 A（Ctsa）是一种丝氨酸蛋白酶，影响干细胞的功能和特性。然而，关于 Ctsa 在干细胞中的作用的研究有限。在这里，我们观察到 mESC 分化过程中 Ctsa 表达水平显著增加。此外，我们建立了 Ctsa 敲低 mESCs。Ctsa 敲低导致 Erk1/2 磷酸化，进而抑制 mESCs 的多能性，并诱导 G2/M 细胞周期阻滞以抑制 mESC 增殖。敲低还导致 mESCs 异常分化和分化标记物异常表达。此外，我们还鉴定了在裸鼠中抑制畸胎瘤形成。我们的结果表明，Ctsa 影响 mESC 的多能性、增殖、细胞周期和分化，并强调了 Ctsa 作为一种核心因子调节各种 mESC 特性的潜力。研究意义：我们的结果表明组织蛋白酶 A（Ctsa）影响 mESCs 的特性。抑制 Ctsa 导致小鼠胚胎干细胞（mESCs）的多能性降低。此外，Ctsa 抑制通过细胞周期阻滞导致增殖减少。此外，Ctsa 抑制降低了 mESCs 分化能力和畸胎瘤形成。我们的结果表明 Ctsa 是控制 mESC 能力的重要因素。
