Tan Xin, Liang Xiaojing, Feng Yi, Xie Ming, Zhong Kun, Luo Wenwu, Wang Yurao, Yin Yu, Cai Yongping
Department of Pathology, School of Basic Medical Science, Anhui Medical University, Hefei, 230032, People's Republic of China.
Department of Pathology, the First Affiliated Hospital of Anhui Medical University, Hefei, 230022, People's Republic of China.
Med Oncol. 2025 Mar 19;42(4):120. doi: 10.1007/s12032-025-02657-w.
Osteosarcoma is a highly invasive and metastatic primary malignant bone tumor, and resistance to chemotherapy remains a major therapeutic challenge. Our previous studies showed that increased Forkhead box protein M1 (FoxM1) expression promotes osteosarcoma progression. While NIMA-related kinase 2 (NEK2) has emerged as a potential oncogenic factor, its functional role and molecular mechanisms in osteosarcoma remain poorly understood. Pearson's correlation analysis was performed to assess the relationship between FoxM1 and NEK2 expression using the GSE33382 dataset from GEO. Coimmunoprecipitation (Co-IP) was employed to investigate FoxM1-NEK2 interactions. NEK2 expression was modulated in the HOS and U2OS osteosarcoma cell lines through pharmacological inhibition (MBM-55), siRNA-mediated knockdown, and plasmid-mediated overexpression. Cellular proliferation was evaluated via CCK-8 and colony formation assays. Transwell migration/invasion assays and flow cytometry were performed to assess the metastatic potential and apoptosis, respectively. The protein levels of FoxM1, NEK2, and AKT/p-AKT were analyzed by Western blotting. Western blot analyses of FoxM1-overexpressing cell lines and RCM-1-treated cells revealed a positive correlation between NEK2 and FoxM1 levels. Co-IP confirmed their interaction. NEK2 knockdown significantly suppressed proliferation, migration, and invasion; enhanced cisplatin sensitivity (reduced the IC); and promoted apoptosis. Conversely, NEK2 overexpression exacerbated malignant phenotypes and decreased chemosensitivity. Mechanistically, NEK2 activation was shown to drive osteosarcoma progression via AKT/p-AKT pathway activation. This study revealed that NEK2 promotes osteosarcoma proliferation, invasion, migration, and chemoresistance while inhibiting apoptosis, likely through AKT/p-AKT signaling. These effects may be regulated by FoxM1.
骨肉瘤是一种具有高度侵袭性和转移性的原发性恶性骨肿瘤，对化疗的耐药性仍然是主要的治疗挑战。我们之前的研究表明，叉头框蛋白M1（FoxM1）表达增加会促进骨肉瘤进展。虽然NIMA相关激酶2（NEK2）已成为一种潜在的致癌因子，但其在骨肉瘤中的功能作用和分子机制仍知之甚少。使用来自基因表达综合数据库（GEO）的GSE33382数据集进行Pearson相关性分析，以评估FoxM1与NEK2表达之间的关系。采用免疫共沉淀（Co-IP）法研究FoxM1-NEK2相互作用。通过药物抑制（MBM-55）、小干扰RNA（siRNA）介导的敲低和质粒介导的过表达，在人骨肉瘤细胞系HOS和U2OS中调节NEK2表达。通过细胞计数试剂盒-8（CCK-8）和集落形成试验评估细胞增殖。分别进行Transwell迁移/侵袭试验和流式细胞术以评估转移潜能和细胞凋亡。通过蛋白质免疫印迹法分析FoxM1、NEK2和AKT/p-AKT的蛋白水平。对FoxM1过表达细胞系和RCM-1处理的细胞进行的蛋白质免疫印迹分析显示，NEK2与FoxM1水平呈正相关。免疫共沉淀证实了它们之间的相互作用。NEK2敲低显著抑制增殖、迁移和侵袭；增强顺铂敏感性（降低半数抑制浓度）；并促进细胞凋亡。相反，NEK2过表达加剧了恶性表型并降低了化疗敏感性。机制上，NEK2激活被证明通过AKT/p-AKT途径激活来驱动骨肉瘤进展。本研究表明，NEK2可能通过AKT/p-AKT信号传导促进骨肉瘤增殖、侵袭、迁移和化疗耐药，同时抑制细胞凋亡。这些作用可能受FoxM1调节。
