TOPK Activation Exerts Protective Effects on Cisplatin-induced Acute Kidney Injury
TOPK Activation Exerts Protective Effects on Cisplatin-induced Acute Kidney Injury
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DOI：
10.1007/s11596-022-2545-0
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发表时间：
2022-06-09
影响因子：
2.4
通讯作者：
Yao, Li-jun
中科院分区：
文献类型：
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作者：
Zhang, Hui;Dong, Qing-qing;Yao, Li-jun
目的T-LAK细胞起源蛋白激酶(TOPK)是一种PSD95-Disc Large-ZO1(PDZ-ZO1)结合蛋白，是丝裂原活化蛋白激酶(MAPK)家族的新成员。研究表明，TOPK在肿瘤细胞的功能中起着关键作用，包括细胞凋亡和有丝分裂。然而，TOPK在顺铂诱导的急性肾损伤(CP-AKI)中的作用尚不清楚。本研究旨在探讨TOPK在CP-AKI中的作用及其机制。方法用C57BL/6小鼠和体外培养的肾小管上皮细胞(TECs)建立CP-AKI小鼠或细胞模型。用TOPK特异性抑制剂OTS514刺激TECs，或用重组激活的TOPK-T9E载体转染TECs，以抑制或激活TOPK。在用OTS514或顺铂刺激后，用AKT抑制剂VIII处理TECs。用Western blotting和流式细胞术检测TECs的细胞周期和细胞凋亡率。结果顺铂在体内和体外均能显著抑制TOPK活性。此外，TOPK的特异性抑制剂OTS514对TOPK的药理抑制作用加剧了顺铂诱导的细胞周期停滞在G2/M期和TECs的凋亡。TOPK-T9E载体介导的TOPK激活可部分逆转顺铂诱导的TEC细胞周期停滞于G2/M期和细胞凋亡。此外，作为TOPK靶蛋白的AKT/蛋白激酶B(PKB)在培养的TECs中被顺铂抑制。药物抑制AKT进一步加重顺铂或TOPK抑制诱导的TECs凋亡。在CP-AKI细胞模型中，TOPK通过AKT途径系统地介导细胞凋亡。结论TOPK激活可能通过改善细胞G2/M期停滞和细胞凋亡而对CP-AKI产生保护作用。
Objective T-LAK-cell-originated protein kinase (TOPK), a PSD95-Disc large-ZO1 (PDZ) binding kinase (PBK), is a novel member of the mitogen-activated protein kinase (MAPK) family. Studies have shown that TOPK plays a critical role in the function of tumor cells, including apoptosis and mitosis. However, little is known on the effect of TOPK in cisplatin-induced acute kidney injury (CP-AKI). This study aimed to investigate the role and mechanism of TOPK in CP-AKI. Methods Cisplatin was administered to C57BL/6 mice and cultured kidney tubular epithelial cells (TECs) to establish the CP-AKI murine or cellular models. TECs were then stimulated with the specific inhibitor of TOPK OTS514 or transfected with the recombinant-activated plasmid TOPK-T9E to inhibit or activate TOPK. The TECs were treated with AKT inhibitor VIII following stimulation with OTS514 or cisplatin. Western blotting and flow cytometry were used to evaluate the cell cycle and apoptosis of TECs. Results The analysis revealed that the TOPK activity was significantly suppressed by cisplatin, both in vivo and in vitro. Furthermore, the pharmacological inhibition of TOPK by OTS514, a specific inhibitor of TOPK, exacerbated the cisplatin-induced cell cycle arrest in the G2/M phase and apoptosis of cultured TECs. Moreover, the TOPK activation via the TOPK-T9E plasmid transfection could partially reverse the cell cycle arrest at the G2/M phase and apoptosis of cisplatin-treated TECs. In addition, AKT/protein kinase B (PKB), as a TOPK target protein, was inhibited by cisplatin in cultured TECs. The pharmaceutical inhibition of AKT further aggravated the apoptosis of TECs induced by cisplatin or TOPK inhibition. TOPK systematically mediated the apoptosis via the AKT pathway in the CP-AKI cell model. Conclusion These results indicate that TOPK activation protects against CP-AKI by ameliorating the G2/M cell cycle arrest and cell apoptosis.