Yang Junhong, Yue Guangcheng, Fan Zhiguo, Zhang Ning, Nie Shiwei, Li Jing, Ji Yuanyuan
Department of Medical Oncology, The Affiliated Anyang Tumor Hospital of Henan University of Science and Technology, Anyang, 455000, China.
Department of Thoracic Surgery, The Affiliated Anyang Tumor Hospital of Henan University of Science and Technology, Anyang, 455000, China.
Drug Dev Res. 2025 Apr;86(2):e70087. doi: 10.1002/ddr.70087.
Lung adenocarcinoma (LUAD) is one of the main causes of death in cancer patients, as its hidden course is difficult to uncover, resulting in many patients being diagnosed as advanced. Late-stage LUAD patients are prone to develop resistance to cisplatin. This study aims to explore the potential molecular regulatory mechanism of NEK2 on cisplatin resistance in LUAD cells. The expression levels of NEK2 and FOXA1 in LUAD tissues were analyzed based on bioinformatics methods. qRT-PCR analysis was carried out to measure the mRNA expression levels of NEK2 and FOXA1 in LUAD cells. CCK8 detected and calculated cell viability and IC values for each group of cells. Gene set enrichment analysis (GSEA) analyzed signaling pathways enriched by the NEK2 gene in LUAD. Dual luciferase and CHIP experiments were conducted to verify the binding relationship between NEK2 and FOXA1. Comet assay was utilized to analyze the level of DNA damage in LUAD cells. Western blot (WB) measured the expression levels of DNA damage-related proteins (γ-H2AX, p-ATM). The experimental results showed that FOXA1 and NEK2 were highly expressed in LUAD tissues and cells. GSEA analysis showed that NEK2 was enriched in DNA damage-related pathways, and silencing NEK2 could reduce the vitality of LUAD cisplatin-resistant cells, lower the IC value of cells to cisplatin, and increase their DNA damage levels. FOXA1 can target the promoter region that binds to NEK2, and it can activate NEK2 through transcription to promote DNA damage repair and cisplatin resistance in cisplatin-resistant LUAD cells. This study confirms that FOXA1 can target NEK2 to promote DNA damage repair and cisplatin resistance in LUAD cells, providing a new valuable therapeutic target for the treatment of LUAD and the control of chemotherapy drug resistance.
肺腺癌（LUAD）是癌症患者死亡的主要原因之一，因其病程隐匿难以发现，导致许多患者被诊断为晚期。晚期LUAD患者容易对顺铂产生耐药性。本研究旨在探讨NEK2对LUAD细胞顺铂耐药性的潜在分子调控机制。基于生物信息学方法分析LUAD组织中NEK2和FOXA1的表达水平。采用qRT-PCR分析检测LUAD细胞中NEK2和FOXA1的mRNA表达水平。CCK8检测并计算每组细胞的活力和IC值。基因集富集分析（GSEA）分析LUAD中NEK2基因富集的信号通路。进行双荧光素酶和CHIP实验以验证NEK2与FOXA1之间的结合关系。利用彗星试验分析LUAD细胞中的DNA损伤水平。蛋白质免疫印迹法（WB）检测DNA损伤相关蛋白（γ-H2AX、p-ATM）的表达水平。实验结果表明，FOXA1和NEK2在LUAD组织和细胞中高表达。GSEA分析表明，NEK2在DNA损伤相关通路中富集，沉默NEK2可降低LUAD顺铂耐药细胞的活力，降低细胞对顺铂的IC值，并增加其DNA损伤水平。FOXA1可靶向与NEK2结合的启动子区域，并通过转录激活NEK2，以促进顺铂耐药LUAD细胞中的DNA损伤修复和顺铂耐药性。本研究证实FOXA1可靶向NEK2以促进LUAD细胞中的DNA损伤修复和顺铂耐药性，为LUAD的治疗及化疗耐药性的控制提供了一个新的有价值的治疗靶点。
