Qin Hai, Yuan Manqin, Yuan Yaqin, Xia Fengqiong, Yang Yonghong
Department of Clinical Laboratory, Beijing Jishuitan Hospital Guizhou Hospital, No. 206, Sixian Street, Baiyun District, Guiyang City, Guizhou Province, China.
Department of Clinical Laboratory Medicine, Guizhou Medical University, No. 9, Beijing Road, Yunyan District, Guiyang City, Guizhou Province, China.
Transl Oncol. 2025 Jan;51:102186. doi: 10.1016/j.tranon.2024.102186. Epub 2024 Nov 4.
Colorectal cancer (CRC) is a prevalent malignancy with poor patient survival, and NIMA-associated kinase 2 (NEK2) has been implicated in the pathogenesis and progression of various cancers, including CRC. This study aimed to investigate the impact of NEK2 on CRC cell functionality and its interaction with the TGF-β/Smad signaling pathway. NEK2 expression in CRC tissues and cell lines was assessed, and its association with patient survival was analyzed. Functional assays, including NEK2 knockdown via lentiviral infection, RT-qPCR, Western blotting, CCK-8 assay, Transwell migration, invasion assays, and goblet cell formation assays, were employed to evaluate NEK2's effects on CRC cell proliferation, migration, invasion, and stemness. Mechanistic studies explored the TGF-β/Smad2 signaling pathway, utilizing co-immunoprecipitation (Co-IP) and protein interaction analyses. In vivo experiments further evaluated NEK2's role in tumor initiation, metastasis, and chemoresistance. NEK2 was found to be upregulated in CRC tissues and correlated with poor survival. NEK2 knockdown inhibited CRC cell behaviors, while NEK2 activated the TGF-β/Smad2 signaling pathway through Smad2/3 phosphorylation. Overexpression of Smad2/3 reversed NEK2 knockdown effects, confirming the importance of this pathway in CRC. In vivo, NEK2 promoted tumor initiation, metastasis, and chemoresistance, effects partially reversed by Smad2/3 overexpression. These findings reveal the critical role of NEK2 in CRC progression and underscore its potential as a therapeutic target, offering new insights into the molecular mechanisms driving CRC and informing targeted therapy development.
结直肠癌（CRC）是一种常见的恶性肿瘤，患者生存率较低，而NIMA相关激酶2（NEK2）已被证实与包括CRC在内的多种癌症的发病机制和进展有关。本研究旨在探讨NEK2对CRC细胞功能的影响及其与转化生长因子-β（TGF-β）/Smad信号通路的相互作用。评估了NEK2在CRC组织和细胞系中的表达，并分析了其与患者生存率的关系。采用包括通过慢病毒感染敲低NEK2、逆转录-定量聚合酶链反应（RT-qPCR）、蛋白质免疫印迹法、细胞计数试剂盒-8（CCK-8）检测、Transwell迁移和侵袭检测以及杯状细胞形成检测等功能实验，以评估NEK2对CRC细胞增殖、迁移、侵袭和干性的影响。机制研究利用免疫共沉淀（Co-IP）和蛋白质相互作用分析探索了TGF-β/Smad2信号通路。体内实验进一步评估了NEK2在肿瘤起始、转移和化疗耐药中的作用。研究发现NEK2在CRC组织中上调，且与较差的生存率相关。敲低NEK2可抑制CRC细胞行为，而NEK2通过Smad2/3磷酸化激活TGF-β/Smad2信号通路。Smad2/3的过表达逆转了NEK2敲低的作用，证实了该信号通路在CRC中的重要性。在体内，NEK2促进肿瘤起始、转移和化疗耐药，Smad2/3过表达可部分逆转这些作用。这些发现揭示了NEK2在CRC进展中的关键作用，并强调了其作为治疗靶点的潜力，为驱动CRC的分子机制提供了新的见解，并为靶向治疗的开发提供了依据。
