Jin Yuzhao, Liao Luyu, Chen Qianping, Tang Bufu, Jiang Jin, Zhu Ji, Bai Minghua
Postgraduate Training Base Alliance of Wenzhou Medical University (Zhejiang Cancer Hospital), Hangzhou, Zhejiang, 310022, China.
Hangzhou Institute of Medicine (HIM), Chinese Academy of Sciences Hangzhou, Hangzhou, 310000, China.
Cancer Cell Int. 2025 Jan 31;25(1):31. doi: 10.1186/s12935-025-03643-y.
Previous studies have found that neutrophil extracellular traps (NETs) are highly expressed in colorectal cancer (CRC) and are associated with poor prognosis. Currently, there are few studies on the relationship between NETs and CRC, so we tried to explore new markers based on NETs to assist in the treatment of CRC.
We jointly screened three major NETs genes through machine learning. Large-sample RNA transcriptome and single-cell transcriptome analysis further confirmed that TIMP1 is a core gene in NETs. We used small interfering RNA to knockdown TIMP1, and verified the ability of TIMP1 in CRC proliferation, invasion and migration through western blot, transwell, cell scratch assay, cell clone formation and other experiments.
We screened out three major NETs Genes: TIMP1, F3, and CRISPLD2 based on machine learning. The NETs score constructed based on this not only predicts the prognosis of CRC patients but also shows significant differences in MSI status, chenckpoints expression, and predicted efficacy of PD-L1 targeted therapy. Transcriptome and single-cell data reveal that TIMP1 is highly expressed in neutrophils and is associated with poor prognosis in colorectal cancer patients and the occurrence of ferroptosis. Biological experiments have proven that TIMP1 can promote the proliferation, invasion and migration of CRC.
Bioinformatics analysis combined with experimental verification showed that TIMP1 is related to ferroptosis and plays a promoting role in the invasion, migration and proliferation of CRC.
既往研究发现中性粒细胞胞外诱捕网（NETs）在结直肠癌（CRC）中高表达且与预后不良相关。目前，关于NETs与CRC关系的研究较少，因此我们试图探索基于NETs的新标志物以辅助CRC的治疗。
我们通过机器学习联合筛选出三个主要的NETs基因。大样本RNA转录组和单细胞转录组分析进一步证实金属蛋白酶组织抑制因子1（TIMP1）是NETs中的核心基因。我们使用小干扰RNA敲低TIMP1，并通过蛋白质免疫印迹法、Transwell实验、细胞划痕实验、细胞克隆形成等实验验证TIMP1在CRC增殖、侵袭和迁移方面的能力。
我们基于机器学习筛选出三个主要的NETs基因：TIMP1、凝血因子Ⅲ（F3）和富含半胱氨酸的分泌蛋白2（CRISPLD2）。基于此构建的NETs评分不仅能预测CRC患者的预后，还在微卫星不稳定性（MSI）状态、检查点表达以及程序性死亡受体1（PD-L1）靶向治疗的预测疗效方面显示出显著差异。转录组和单细胞数据显示TIMP1在中性粒细胞中高表达，与结直肠癌患者的不良预后及铁死亡的发生相关。生物学实验证明TIMP1可促进CRC的增殖、侵袭和迁移。
生物信息学分析结合实验验证表明，TIMP1与铁死亡相关，在CRC的侵袭、迁移和增殖中起促进作用。
