Accumulative evidence shows that an organoid is a more practical and reliable tool in cancer biology research. This study aimed to identify and validate crucial genes involved in non-small cell lung cancer carcinogenesis and development using the transcriptomic analysis of tumor tissues and organoids. Gene set enrichment analysis (GSEA) of tumor tissues, tumor organoids, and normal tissues was performed to reveal the similar and different mechanisms involved in lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC) carcinogenesis and progression. Differentially expressed gene analysis, prognostic analysis, and gene co-expression network analysis were further used to identify hub genes involved in LUAD and LUSC carcinogenesis and development. Finally, LUAD cell lines and organoids were used to validate these findings. GSEA analysis was performed to reveal the similar mechanisms involved in LUAD and LUSC carcinogenesis and development, such as P53 signaling pathway, base mismatch repair, DNA replication, cAMP signaling pathway and PPAR pathway. However, comparing with LUSC organoids, LUAD organoids showed downregulation of immune-related pathways, inflammation-related pathways, MAPK signaling pathways, and Rap1 signaling pathways, although these pathways were downregulated in LUAD and LUSC tissues by comparing with normal lung tissues. Further gene co-expression network analysis and prognostic analysis indicated CDK1, CCNB2, and CDC25A as the hub tumor-promoting genes in LUAD but not in LUSC, which were further validated in other datasets. Using LUAD cell lines and organoid models, CDK1 and CCNB2 knockdown were found to suppress LUAD proliferation. However, CDC25A knockdown did not inhibit LUAD cell line proliferation but could effectively suppress LUAD organoid growth, indicating that an organoid could be used as an effective tool to study cancer biology in LUAD. The results revealed CDK1, CCNB2, and CDC25A as the hub genes involved in LUAD carcinogenesis and development, which could be used as the potential biomarkers and targets for LUAD.

中文翻译：

---

肿瘤组织和类器官的转录组学分析揭示了调控肺腺癌增殖的关键基因

越来越多的证据表明，类器官是癌症生物学研究中更实用、更可靠的工具。本研究旨在利用肿瘤组织和类器官的转录组学分析来鉴定和验证与非小细胞肺癌致癌作用和发展相关的关键基因。对肿瘤组织、肿瘤类器官和正常组织进行基因集富集分析 (GSEA) 以揭示参与肺腺癌 (LUAD) 和肺鳞状细胞癌 (LUSC) 癌变和进展的相似和不同机制。进一步使用差异表达基因分析、预后分析和基因共表达网络分析来鉴定参与 LUAD 和 LUSC 癌变和发展的中枢基因。最后，使用 LUAD 细胞系和类器官来验证这些发现。进行GSEA分析以揭示参与LUAD和LUSC致癌和发展的相似机制，例如P53信号通路、碱基错配修复、DNA复制、cAMP信号通路和PPAR通路。然而，与 LUSC 类器官相比，LUAD 类器官显示免疫相关通路、炎症相关通路、MAPK 信号通路和 Rap1 信号通路的下调，尽管与正常肺组织相比，这些通路在 LUAD 和 LUSC 组织中被下调。进一步的基因共表达网络分析和预后分析表明 CDK1、CCNB2 和 CDC25A 是 LUAD 而非 LUSC 中的中枢促肿瘤基因，这在其他数据集中得到了进一步验证。使用 LUAD 细胞系和类器官模型，发现 CDK1 和 CCNB2 敲低可抑制 LUAD 增殖。然而，CDC25A 敲低不会抑制 LUAD 细胞系增殖，但可以有效抑制 LUAD 类器官的生长，表明类器官可作为研究 LUAD 中癌症生物学的有效工具。结果显示CDK1、CCNB2和CDC25A是参与LUAD致癌和发展的枢纽基因，可作为LUAD的潜在生物标志物和靶点。