癌睾抗原（CTAs）常表达于肿瘤和睾丸组织，但不表达于其他正常组织。迄今为止，还没有一项针对与子宫内膜癌（EC）发展相关的CTA基因表达和临床意义的全面研究。此外，CTA基因TTK蛋白激酶（TTK）在EC中的临床相关性、生物学角色和分子机制尚未被完全理解。我们全面调查了通过生物信息学方法分析TCGA数据库中的EC样本中与TTK过度表达相关的基因组、转录组和表观遗传学改变。我们还使用来自GEO数据库的EC样本的单细胞数据，进一步研究了与TTK失调相关的低生存机制。细胞功能实验用于确认TTK在EC细胞中的生物学作用。我们确定了80种CTA基因在EC中比正常组织更丰富表达，并且TTK的高表达与EC患者的低生存率显著相关。此外，ROC分析显示，TTK可以准确区分I期EC组织和良性子宫内膜样本，表明TTK具有成为早期EC检测生物标志物的潜力。我们发现TTK过度表达在高级别、晚期肿瘤、浆液性癌和TP53改变的EC患者中更为普遍。此外，在EC组织中，TTK表达与EMT相关基因呈强阳性相关性。在单细胞转录组数据中，我们确认了一种高表达TTK和已知上皮间充质转化（EMT）相关基因和转录因子的增殖细胞亚群。当增殖细胞根据TTK表达水平分组时，TTK高组中过表达的基因显示出在控制化疗耐药性方面的功能参与。利用shRNA抑制EC细胞中的TTK表达导致细胞增殖、侵袭、EMT和化疗耐药性均大幅度下降。进一步研究确定了microRNA-21（miR-21）作为TTK诱导的EMT和化疗耐药性的重要下游调节因子。最后，TTK抑制剂AZ3146在减少EC细胞生长和侵袭，并增强由紫杉醇引起的EC细胞凋亡方面具有良好效果。我们的研究结果确定了TTK作为EC的新生物标记物和尚未知的致癌功能的临床意义。本研究建议，TTK的治疗靶向可能为EC治疗提供可行途径。©2023年作者。

Cancer-testis antigens (CTAs) are often expressed in tumor and testicular tissues but not in other normal tissues. To date, there has been no comprehensive study of the expression and clinical significance of CTA genes associated with endometrial cancer (EC) development. Additionally, the clinical relevance, biological role, and molecular mechanisms of the CTA gene TTK protein kinase (TTK) in EC are yet to be fully understood.Using bioinformatics methods, we comprehensively investigated the genomic, transcriptomic, and epigenetic changes associated with aberrant TTK overexpression in EC samples from the TCGA database. We further investigated the mechanisms of the lower survival associated with TTK dysregulation using single-cell data of EC samples from the GEO database. Cell functional assays were used to confirm the biological roles of TTK in EC cells.We identified 80 CTA genes that were more abundant in EC than in normal tissues, and high expression of TTK was significantly linked with lower survival in EC patients. Furthermore, ROC analysis revealed that TTK could accurately distinguish stage I EC tissues from benign endometrial samples, suggesting that TTK has the potential to be a biomarker for early EC detection. We found TTK overexpression was more prevalent in EC patients with high-grade, advanced tumors, serous carcinoma, and TP53 alterations. Furthermore, in EC tissue, TTK expression showed a strong positive correlation with EMT-related genes. With single-cell transcriptome data, we identified a proliferative cell subpopulation with high expression of TTK and known epithelial-mesenchymal transition (EMT)-related genes and transcription factors. When proliferative cells were grouped according to TTK expression levels, the overexpressed genes in the TTKhigh group were shown to be functionally involved in the control of chemoresistance. Utilizing shRNA to repress TTK expression in EC cells resulted in substantial decreases in cell proliferation, invasion, EMT, and chemoresistance. Further research identified microRNA-21 (miR-21) as a key downstream regulator of TTK-induced EMT and chemoresistance. Finally, the TTK inhibitor AZ3146 was effective in reducing EC cell growth and invasion and enhancing the apoptosis of EC cells generated by paclitaxel.Our findings establish the clinical significance of TTK as a new biomarker for EC and an as-yet-unknown carcinogenic function. This present study proposes that the therapeutic targeting of TTK might provide a viable approach for the treatment of EC.© 2023. The Author(s).