背景:膀胱癌(BLCA)是一种常见且致命的泌尿系癌症,需要有效的治疗方法,特别是肌肉浸润性膀胱癌(MIBC)。细胞周期抑制剂有望恢复BLCA细胞中细胞周期的控制,但他们的临床预后评估是有限的。
方法:转录组和scRNA-seq数据来自癌症基因组图谱计划(TCGA)-BLCA和GSE190888队列,分别。使用R软件和Seurat软件包进行数据分析,包括细胞质量控制,降维,和差异表达基因的鉴定。与细胞周期相关的基因从基因卡网站获得,使用cytoscape软件进行蛋白质-蛋白质相互作用网络分析。功能富集分析,免疫浸润分析,药物敏感性分析,和分子对接使用各种工具和包装进行。培养BLCA细胞系并转染用于体外实验测定,包括RT-qPCR分析,和CCK-8细胞活力测定。
结果:我们确定了32个基因作为BLCA预测的独立风险或保护因子。功能富集分析显示它们参与细胞周期调控,凋亡,和各种信号通路。利用这些基因,我们开发了一个预测BLCA生存的列线图,在BLCA患者中显示出较高的预后分层疗效。确定了四个细胞周期相关的关键基因,包括NCAM1,HBB,CKD6和CTLA4。我们还确定了BLCA患者的主要细胞类型,并根据关键基因的表达水平研究了上皮细胞之间的功能差异。此外,我们观察到癌相关成纤维细胞浸润与风险评分值之间存在高度正相关关系.最后,我们进行了体外实验,以证明NCAM1在BLCA细胞增殖中的抑制作用。
结论:这些研究结果表明,细胞周期相关基因可作为预测BLCA预后的潜在生物标志物,并可能成为开发更有效疗法的治疗靶点。希望,这些发现从细胞周期的角度为BLCA的分子机制和潜在治疗靶点提供了有价值的见解。此外,NCAM1是BLCA癌变中的一种新型细胞增殖抑制因子。
BACKGROUND: Bladder cancer (BLCA) is a prevalent and deadly form of urinary cancer, and there is a need for effective therapies, particularly for muscle-invasive bladder cancer (MIBC). Cell cycle inhibitors show promise in restoring control of the cell cycle in BLCA cells, but their clinical prognosis evaluation is limited.
METHODS: Transcriptome and scRNA-seq data were collected from the Cancer Genome Atlas Program (TCGA)-BLCA and GSE190888 cohort, respectively. R software and the Seurat package were used for data analysis, including cell quality control, dimensionality reduction, and identification of differentially expressed genes. Genes related to the cell cycle were obtained from the genecards website, and a protein-protein interaction network analysis was performed using cytoscape software. Functional enrichment analysis, immune infiltration analysis, drug sensitivity analysis, and molecular docking were conducted using various tools and packages. BLCA cell lines were cultured and transfected for in vitro experimental assays, including RT-qPCR analysis, and CCK-8 cell viability assays.
RESULTS: We identified 32 genes as independent risk or protective factors for BLCA prediction. Functional enrichment analysis revealed their involvement in cell cycle regulation, apoptosis, and various signaling pathways. Using these genes, we developed a nomogram for predicting BLCA survival, which displayed high prognosis stratification efficacy in BLCA patients. Four cell cycle associated key genes identified, including
NCAM1, HBB, CKD6, and CTLA4. We also identified the main cell types in BLCA patients and investigated the functional differences between epithelial cells based on their expression levels of key genes. Furthermore, we observed a high positive correlative relationship between the infiltration of cancer-associated fibroblasts and the risk score value. Finally, we conducted in vitro experiments to demonstrate the suppressive role of
NCAM1 in BLCA cell proliferation.
CONCLUSIONS: These findings suggest that cell cycle associated genes could serve as potential biomarkers for predicting BLCA prognosis and may represent therapeutic targets for the development of more effective therapies. Hopefully, these findings provide valuable insights into the molecular mechanisms and potential therapeutic targets in BLCA from the perspective of cell cycle. Moreover,
NCAM1 was a novel cell proliferation suppressor in the BLCA carcinogenesis.