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| Identification of tumor microenvironment and intercellular communication-related signaling molecules of castration-resistant prostate cancer based on single cell RNA sequencing |
| ZHU Xiaoren 1,2, CHEN Minbin 1,2, JIANG Jianzhuo 2,3, XUE Mengen 4, LIU Yuanyuan 2,3 |
| (1. Department of Radiotherapy and Oncology, Affiliated Kunshan Hospital of Jiangsu University, Kunshan Jiangsu 215300; 2. School of Medicine, Jiangsu University, Zhenjiang Jiangsu 212013; 3. Clinical Research & Lab Center, Affiliated Kunshan Hospital of Jiangsu University, Kunshan Jiangsu 215300; 4. Gusu School, Nanjing Medical University, Suzhou Jiangsu 215000, China） |
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Abstract Objective： To explore intercellular communication-related signaling molecules in tumor microenvironment (TME) of castration-resistant prostate cancer (CRPC) and analyze their relationship with tumor occurrence and development. Methods： The main components of the TME in CRPC was analyzed using singlecell RNA sequencing (scRNA-seq) data (GSE137829) from CRPC patients found in the Gene Expression Omnibus (GEO) database. CellPhoneDB was used to examine interactions between different cells within the microenvironment, with a focus on revealing the connections between cancer-associated fibroblasts (CAFs) and malignant epithelial cells. CellChat was applied to analyze signaling molecules involved in communication between CAFs and malignant epithelial cells. Data from The Cancer Genome Atlas Prostate Adenocarcinoma (TCGA-PRAD) were integrated to identify signaling molecules closely related to the prognosis of CRPC patients. siRNA was used to knock down the expression of the signaling molecule Delta-like canonical Notch ligand 3 (DLL3) in CRPC cells, and the knockdown efficiency was validated by real-time quantitative PCR and Western blotting. The effects of DLL3 on the biological behaviors of CRPC cells, including proliferation, migration, and invasion, were investigated using CCK8 assays, EdU assays, and Transwell assays with Matrigel, respectively. Singlegene differential analysis, overrepresentation analysis (ORA), and gene set enrichment analysis (GSEA) were also performed to identify pathways enriched with DLL3-related differential expression genes. Results： The results of single-cell transcriptomic sequencing indicated that the main cellular components of the TME in CRPC include malignant epithelial cells, CAFs and immune cells. Close communication between CAFs and malignant epithelial cells was mediated by interaction-related signaling molecules, with 63 signaling molecules identified as closely related to the communication between CAFs and malignant epithelial cells. Univariate Cox regression analysis identified key signaling molecules, including tumor necrosis factorrelated ligand superfamily member 12 (TNFSF12) and DLL3, whose transcriptional expression were significantly associated with the prognosis of CRPC patients. Notably, high expression of DLL3 was significantly associated with poor prognosis in CRPC patients. In vitro experimental results demonstrated that knockdown of DLL3 significantly inhibited the growth, proliferation, migration and invasion capabilities of CRPC cells. KEGG and GSEA enrichment analyses indicated that differentially expressed genes related to DLL3 were significantly enriched in metabolic pathways. Conclusion： In the TME, CAFs and malignant epithelial cells communicate closely through intercellular signaling molecules. Among these signaling molecules, the expression of DLL3 is negatively correlated with cancer prognosis and affects the biological behavior of CRPC cells, suggesting that DLL3 may be a key signaling molecule in the malignant progression of human CRPC.
［Key words］castration-resistant prostate cancer; scRNA-seq; tumor microenvironment; cell communication; DLL3
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Received: 16 January 2024
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. [J]. Journal of Jiangsu University(Medicine Edition), 2024, 34(04): 290-. |
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