Department of Ophthalmology, Hallym University College of Medicine, 1, Hallymdaehak-gil, Chuncheon-si, Gangwon-do 24252, Korea.
Department of Ophthalmology, Hallym University Medical Center, 1 Shingil-ro, Youngdeungpo-gu, Seoul 07441, Korea.
Int J Mol Sci. 2020 Jun 20;21(12):4397. doi: 10.3390/ijms21124397.
Human corneal endothelial cells (hCECs) pump out water from the stroma and maintain the clarity of the cornea. The sex-determining region Y-box 2 (SOX2) participates in differentiation during the development of the anterior segment of the eye and is found in the periphery of wounded corneas. This study was performed to investigate the effect of SOX2 repression on hCECs.
Cultured hCECs were transfected by siRNA for . The wound healing rate and cell viability were measured. The cell proliferation-associated protein level was evaluated by Western blotting and RT-PCR. The energy production and mitochondrial function were measured, and cell shape and WNT signaling were assessed.
Upon transfecting the cultured cells with siRNA for the SOX2 level was reduced by 80%. The wound healing rate and viability were also reduced. Additionally, CDK1, cyclin D1, SIRT1, and ATP5B levels were reduced, and CDKN2A and pAMPK levels were increased. Mitochondrial oxidative stress and mitochondrial viability decreased, and the cell shape became elongated. Furthermore, SMAD1, SNAI1, WNT3A, and β-catenin levels were increased.
SOX2 repression disrupts the normal metabolism of hCECs through modulating WNT signaling and mitochondrial functions.
人眼角膜内皮细胞（hCECs）从基质中泵出水，维持角膜的透明度。性别决定区 Y 框 2（SOX2）参与眼前段的分化，存在于受伤角膜的周边。本研究旨在探讨 SOX2 抑制对 hCECs 的影响。
通过 siRNA 转染培养的 hCECs。测量伤口愈合率和细胞活力。通过 Western blot 和 RT-PCR 评估与细胞增殖相关的蛋白水平。测量能量产生和线粒体功能，并评估细胞形态和 WNT 信号。
用 siRNA 转染培养细胞后，SOX2 水平降低了 80%。伤口愈合率和活力也降低了。此外，CDK1、cyclin D1、SIRT1 和 ATP5B 水平降低，CDKN2A 和 pAMPK 水平升高。线粒体氧化应激和线粒体活力降低，细胞形态变长。此外，SMAD1、SNAI1、WNT3A 和 β-catenin 水平升高。
SOX2 抑制通过调节 WNT 信号和线粒体功能破坏 hCECs 的正常代谢。
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