IntroductionBladder 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.MethodsTranscriptome 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.ResultsWe 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.ConclusionThese 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.

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从细胞周期 NCAM1 角度对膀胱癌进行单细胞和批量分辨率的综合分析

简介膀胱癌（BLCA）是一种常见且致命的泌尿系统癌症，需要有效的治疗方法，特别是对于肌层浸润性膀胱癌（MIBC）。细胞周期抑制剂显示出恢复 BLCA 细胞细胞周期控制的希望，但其临床预后评估有限。方法转录组和 scRNA-seq 数据分别从癌症基因组图谱计划 (TCGA)-BLCA 和 GSE190888 队列中收集。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识别。从genecards网站获得与细胞周期相关的基因，并使用cytoscape软件进行蛋白质-蛋白质相互作用网络分析。使用各种工具和软件包进行功能富集分析、免疫浸润分析、药物敏感性分析和分子对接。培养并转染 BLCA 细胞系用于体外实验测定，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素。功能富集分析揭示了它们参与细胞周期调节、细胞凋亡和各种信号通路。利用这些基因，我们开发了一个用于预测 BLCA 存活率的列线图，该列线图在 BLCA 患者中显示出较高的预后分层功效。鉴定出四个细胞周期相关关键基因，包括 NCAM1、HBB、CKD6 和 CTLA4。我们还鉴定了 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，我们观察到癌症相关成纤维细胞的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来证明 NCAM1 在 BLCA 细胞增殖中的抑制作用。结论这些发现表明细胞周期相关基因可以作为预测 BLCA 预后的潜在生物标志物，并可能代表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有价值的见解。此外，NCAM1 是 BLCA 致癌过程中一种新型细胞增殖抑制因子。