Wen Ge, Niu Shaoqing, Mei Shiqi, Wang Senming
Department of Oncology, Zhujiang Hospital, Southern Medical University, Guangzhou, 510280, China.
Department of Radiation Oncology, Guangdong Provincial Key Laboratory of Major Obstetric Diseases, Guangdong Provincial Clinical Research Center for Obstetrics and Gynecology, The Third Affiliated Hospital, Guangzhou Medical University, Guangzhou, 510150, China.
Sci Rep. 2025 May 8;15(1):16085. doi: 10.1038/s41598-025-00585-3.
Lung adenocarcinoma (LUAD) is the most popular lung cancer type with highly mortality. We performed a single cell RNA-seq analysis to explore characteristic of cancer stem cells in LUAD. We downloaded the single cell RNA-seq data (GSE149655) from the GEO database, the scRNA-seq analysis was performed by using the "Seurat" and "harmony" R package. The FindMarkers function and "ClusterProlifer" package was used for differentially expressed genes (DEGs) and function enrichment analysis. The protein-protein interaction and transcriptional regulatory network were performed by STRING and ChIPBase database. Immunohistochemistry tests to be used to observe differences in the expression of specific genes in LUAD and paracancerous tissue samples. BEAS-2B and A549 cells was used for vitro assay and the qRT-PCR, western blotting, wound healing, trans-well assays, EdU tests, and flow cytometry were performed. A total of 9 cell clusters were obtained after scRNA-seq analysis, in which the cancer stem cells had higher proportion in LUAD samples. Subsequently, function enrichment analysis revealed that the amino sugar and nucleotide sugar metabolism and DNA replication pathways were activated in cancer stem cells (CSCs), which were further sub-divided into 3 subtypes, the LGR5 + stem cell is a major contributor to cancer progression, its hub genes, such as HLA-DPB1, CD74, CTSH and HLA-DRB5 mediated the unique transcriptional state. In addition, the marker genes of three CSCs were also overexpressed in LUAD cells and the CXCL3 played an important role in mediating cell proliferation, apoptosis, migration and invasion of tumor. We performed a scRNA-seq analysis and identified the LGR5 + stem cell as a major contributor in LUAD progression, our findings are expected to provide new insights into the pathogenesis of LUAD.
肺腺癌（LUAD）是最常见且死亡率极高的肺癌类型。我们进行了单细胞RNA测序分析，以探究LUAD中癌症干细胞的特征。我们从GEO数据库下载了单细胞RNA测序数据（GSE149655），使用“Seurat”和“harmony”R包进行scRNA-seq分析。利用FindMarkers函数和“ClusterProlifer”包进行差异表达基因（DEG）和功能富集分析。通过STRING和ChIPBase数据库构建蛋白质-蛋白质相互作用和转录调控网络。采用免疫组织化学检测观察LUAD和癌旁组织样本中特定基因表达的差异。使用BEAS-2B和A549细胞进行体外实验，并进行qRT-PCR、蛋白质免疫印迹、伤口愈合、Transwell实验、EdU检测和流式细胞术。scRNA-seq分析后共获得9个细胞簇，其中癌症干细胞在LUAD样本中的比例更高。随后的功能富集分析表明，氨基糖和核苷酸糖代谢以及DNA复制途径在癌症干细胞（CSC）中被激活，这些细胞进一步细分为3个亚型，LGR5 +干细胞是癌症进展的主要贡献者，其枢纽基因如HLA-DPB1、CD74、CTSH和HLA-DRB5介导了独特的转录状态。此外，三种CSC的标记基因在LUAD细胞中也过表达，且CXCL3在介导肿瘤细胞增殖、凋亡、迁移和侵袭中起重要作用。我们进行了scRNA-seq分析，并确定LGR5 +干细胞是LUAD进展的主要贡献者，我们的研究结果有望为LUAD的发病机制提供新的见解。
