An Jiayue, Song Wei, Wang Qin, Tan Boyu, Fei Xuan, Wang Ruoxi, Li Siyan, Lu Xiyu, Li Youjie, Xie Ning
Department of Clinical Laboratory, The Second Medical College of Binzhou Medical University, Yantai, Shandong 264100, P.R. China.
Department of Biochemistry and Molecular Biology, Binzhou Medical University, Yantai, Shandong 264003, P.R. China.
Oncol Lett. 2025 May 19;30(1):353. doi: 10.3892/ol.2025.15099. eCollection 2025 Jul.
Lung adenocarcinoma (LUAD) is among the most prevalent malignancies worldwide. Cuproptosis, a copper-induced form of cell death, has been identified as a key process in LUAD progression; however, the molecular mechanisms underlying cuproptosis in LUAD and potential therapeutic targets remain unclear. The present study utilized The Cancer Genome Atlas database to retrieve mRNA expression profiles and clinical information of LUAD, identifying 10 candidate genes from differentially expressed genes associated with cuproptosis. Protein-protein interaction analysis indicated that CDK inhibitor 2A (CDKN2A), an upregulated gene in LUAD, may function as a hub gene. Furthermore, multiple online databases were used to analyze Spi-1 proto-oncogene (SPI1), a transcription factor upstream of CDKN2A, which was downregulated in LUAD cuproptosis. The LinkedOmics database identified the p53-mediated cuproptosis-related pathway regulated by CDKN2A. Gene expression patterns were examined through Gene Expression Profiling Interactive Analysis, the Human Protein Atlas and reverse transcription-quantitative polymerase chain reaction. Prognostic significance was assessed using the UALCAN and Kaplan-Meier plotter databases. experiments demonstrated that CDKN2A knockdown and SPI1 overexpression inhibited the proliferation and migration of the H1975 cell line. After copper-induced cuproptosis in H1975 cells, SPI1 expression was upregulated, whereas CDKN2A expression was downregulated. When H1975 cells were pretreated with tetrathiomolybdate, the upregulation of SPI1 was inhibited and the downregulation of CDKN2A was also suppressed. Cell Counting Kit-8 assays indicated that SPI1 overexpression and CDKN2A knockdown facilitated elesclomol-CuCl-induced cuproptosis. Western blot analysis revealed an inverse association between SPI1 overexpression and CDKN2A/p53 levels. In conclusion, the present study demonstrated the role of the SPI1/CDKN2A/p53 axis in LUAD cuproptosis, providing insights into potential therapeutic targets and contributing to clinical research on treatment strategies.
肺腺癌（LUAD）是全球最常见的恶性肿瘤之一。铜死亡是一种由铜诱导的细胞死亡形式，已被确定为LUAD进展中的关键过程；然而，LUAD中铜死亡的分子机制和潜在治疗靶点仍不清楚。本研究利用癌症基因组图谱数据库检索LUAD的mRNA表达谱和临床信息，从与铜死亡相关的差异表达基因中鉴定出10个候选基因。蛋白质-蛋白质相互作用分析表明，LUAD中上调的基因细胞周期蛋白依赖性激酶抑制剂2A（CDKN2A）可能作为枢纽基因发挥作用。此外，使用多个在线数据库分析CDKN2A上游的转录因子Spi-1原癌基因（SPI1），其在LUAD铜死亡中表达下调。LinkedOmics数据库确定了由CDKN2A调节的p53介导的铜死亡相关途径。通过基因表达谱交互式分析、人类蛋白质图谱和逆转录-定量聚合酶链反应检测基因表达模式。使用UALCAN和Kaplan-Meier绘图仪数据库评估预后意义。实验表明，敲低CDKN2A和过表达SPI1可抑制H1975细胞系的增殖和迁移。在H1975细胞中诱导铜死亡后，SPI1表达上调，而CDKN2A表达下调。当H1975细胞用四硫代钼酸盐预处理时，SPI1的上调受到抑制，CDKN2A的下调也受到抑制。细胞计数试剂盒-8检测表明，过表达SPI1和敲低CDKN2A促进了依斯氯铵-氯化铜诱导的铜死亡。蛋白质印迹分析显示SPI1过表达与CDKN2A/p53水平呈负相关。总之，本研究证明了SPI1/CDKN2A/p53轴在LUAD铜死亡中的作用，为潜在治疗靶点提供了见解，并有助于治疗策略的临床研究。
