Zhang Weidong, Ni Yingchen, Li Jianxin, Hua Runjia, Wang Yudong, Yang Huilin, Li Xuefeng, Gan Minfeng, Chu Genglei
Department of Orthopaedic Surgery, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China.
Department of Orthopaedic Surgery, Affiliated Hospital of Nantong University, Nantong, Jiangsu, China.
Heliyon. 2024 Jul 4;10(13):e34127. doi: 10.1016/j.heliyon.2024.e34127. eCollection 2024 Jul 15.
NUAK2 is a member of the AMP-activated protein kinase (AMPK) family, which plays an essential role in cellular processes such as apoptosis, proliferation, and cell fate. Recent studies have already shown that silencing of NUAK2 blocks proliferation and promotes apoptosis of human melanoma cells and liver cancer cells. In addition, NUAK2 is involved in the development of glioblastoma via regulating the expression of cancer stem cell-related genes, and it promotes the cell cycle entry in the glioblastoma cells. However, the expression and the role of NUAK2 in the progress of peripheral nerve regeneration after injury are yet to be elucidated. We observed that NUAK2 was upregulated following distal sciatic nerve crush (SNC). Interestingly, we discovered that NUAK2 showed co-localization with S100 (Schwann cell marker). Furthermore, we found that the NUAK2 had a spatiotemporal protein expression, which was consistent with proliferating cell nuclear-antigen (PCNA). The protein level of NUAK2 and YAP was upregulated in the model of TNF-α-induced Schwann cell (SC) proliferation. Furthermore, flow cytometry analysis, CCK-8, transwell assays, and wound healing assays were all performed with the purpose of exploring the role of NUAK2 in the regulation of SC proliferation and migration. More importantly, we found that NUAK2-deficient SCs showed significantly reduced expression of Yes-associated protein (YAP). Bioinformatic analysis identified upstream regulators of NUAK2 and NUAK2-associated genes (e.g., YAP1). Finally, we investigated the recovery changes during regeneration progress through the walking track analysis. Thus, we speculated that NUAK2 was involved in biochemical and physiological responses of SCs after SNC via YAP-driven proliferation and migration, and this study determined the importance of NUAK2 as a potential target in peripheral nerve regeneration.
NUAK2是AMP激活的蛋白激酶（AMPK）家族的成员，在细胞凋亡、增殖和细胞命运等细胞过程中起着至关重要的作用。最近的研究已经表明，沉默NUAK2会阻碍人黑色素瘤细胞和肝癌细胞的增殖并促进其凋亡。此外，NUAK2通过调节癌症干细胞相关基因的表达参与胶质母细胞瘤的发展，并促进胶质母细胞瘤细胞进入细胞周期。然而，NUAK2在损伤后周围神经再生过程中的表达及作用尚待阐明。我们观察到坐骨神经远端挤压伤（SNC）后NUAK2表达上调。有趣的是，我们发现NUAK2与S100（雪旺细胞标志物）共定位。此外，我们发现NUAK2具有时空蛋白表达，这与增殖细胞核抗原（PCNA）一致。在TNF-α诱导雪旺细胞（SC）增殖的模型中，NUAK2和YAP的蛋白水平上调。此外还进行了流式细胞术分析、CCK-8检测、Transwell检测和伤口愈合检测，以探索NUAK2在调节SC增殖和迁移中的作用。更重要的是，我们发现缺乏NUAK2的SCs中Yes相关蛋白（YAP）的表达显著降低。生物信息学分析确定了NUAK2的上游调节因子和与NUAK2相关的基因（如YAP1）。最后，我们通过行走轨迹分析研究了再生过程中的恢复变化。因此，我们推测NUAK2通过YAP驱动的增殖和迁移参与了SNC后SCs的生化和生理反应，并且本研究确定了NUAK2作为周围神经再生潜在靶点的重要性。
