Identification of CTSC-driven progression in ESCC by single-cell sequencing and experimental validation
BackgroundThe progression of cancer cells is influenced by the tumor microenvironment (TME); however, the molecular mechanisms driving the progression of esophageal squamous cell carcinoma (ESCC) remain unclear. Therefore, we aimed to investigate the TME of ESCC and construct a risk signature based...
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| Format: | Article |
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Frontiers Media S.A.
2025-07-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2025.1585139/full |
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| author | Xin Sui Yongxu Jia Jing Li Jiayao Xu Wenjia Wang Yanru Qin |
| author_facet | Xin Sui Yongxu Jia Jing Li Jiayao Xu Wenjia Wang Yanru Qin |
| author_sort | Xin Sui |
| collection | DOAJ |
| description | BackgroundThe progression of cancer cells is influenced by the tumor microenvironment (TME); however, the molecular mechanisms driving the progression of esophageal squamous cell carcinoma (ESCC) remain unclear. Therefore, we aimed to investigate the TME of ESCC and construct a risk signature based on apoptosis-related genes to identify prognosis-related genes in ESCC.MethodsWe integrated a total of 92,714 cells from 18 samples across three single-cell datasets to analyze the differences in cellular landscapes between primary tumor tissues and adjacent normal tissues. Subsequently, univariate COX regression analysis was employed to construct an apoptosis-related prognostic risk model. The expression of key risk genes was elucidated using immunohistochemistry (IHC). Additionally, the effects of CTSC knockdown on ESCC cell behavior were validated through in vitro and in vivo experiments.ResultsWe identified three malignant cell subtypes (Malig1, Malig2, and Malig4) associated with worse prognosis, which were enriched in apoptosis-related pathways. Pseudotime analysis revealed that the expression scores of apoptosis-related pathways increased along the inferred pseudotime, indicating that apoptosis plays a critical regulatory role in the differentiation of malignant epithelial cells. Furthermore, analysis of the TME demonstrated that immune cells and cancer-associated fibroblasts (CAFs) were significantly more abundant in tumor tissues compared to non-tumor tissues. Additionally, we identified eight apoptosis-related genes associated with prognosis, among which the expression of CTSC was closely correlated with resistance outcomes in patients receiving neoadjuvant immunotherapy. In vitro experiments showed that knockdown of CTSC inhibited the proliferation, migration, and other processes of ESCC cells. In vitro experiments showed that knockdown of CTSC inhibited tumor growth and expression of fibroblast markers.ConclusionsCTSC plays a crucial role in driving TME remodeling and the progression of drug resistance in ESCC, making it a potential target for clinical therapy. |
| format | Article |
| id | doaj-art-9136964fb79542da8f2cd3a69dc02ca0 |
| institution | DOAJ |
| issn | 1664-3224 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj-art-9136964fb79542da8f2cd3a69dc02ca02025-08-20T03:13:32ZengFrontiers Media S.A.Frontiers in Immunology1664-32242025-07-011610.3389/fimmu.2025.15851391585139Identification of CTSC-driven progression in ESCC by single-cell sequencing and experimental validationXin SuiYongxu JiaJing LiJiayao XuWenjia WangYanru QinBackgroundThe progression of cancer cells is influenced by the tumor microenvironment (TME); however, the molecular mechanisms driving the progression of esophageal squamous cell carcinoma (ESCC) remain unclear. Therefore, we aimed to investigate the TME of ESCC and construct a risk signature based on apoptosis-related genes to identify prognosis-related genes in ESCC.MethodsWe integrated a total of 92,714 cells from 18 samples across three single-cell datasets to analyze the differences in cellular landscapes between primary tumor tissues and adjacent normal tissues. Subsequently, univariate COX regression analysis was employed to construct an apoptosis-related prognostic risk model. The expression of key risk genes was elucidated using immunohistochemistry (IHC). Additionally, the effects of CTSC knockdown on ESCC cell behavior were validated through in vitro and in vivo experiments.ResultsWe identified three malignant cell subtypes (Malig1, Malig2, and Malig4) associated with worse prognosis, which were enriched in apoptosis-related pathways. Pseudotime analysis revealed that the expression scores of apoptosis-related pathways increased along the inferred pseudotime, indicating that apoptosis plays a critical regulatory role in the differentiation of malignant epithelial cells. Furthermore, analysis of the TME demonstrated that immune cells and cancer-associated fibroblasts (CAFs) were significantly more abundant in tumor tissues compared to non-tumor tissues. Additionally, we identified eight apoptosis-related genes associated with prognosis, among which the expression of CTSC was closely correlated with resistance outcomes in patients receiving neoadjuvant immunotherapy. In vitro experiments showed that knockdown of CTSC inhibited the proliferation, migration, and other processes of ESCC cells. In vitro experiments showed that knockdown of CTSC inhibited tumor growth and expression of fibroblast markers.ConclusionsCTSC plays a crucial role in driving TME remodeling and the progression of drug resistance in ESCC, making it a potential target for clinical therapy.https://www.frontiersin.org/articles/10.3389/fimmu.2025.1585139/fullesophageal squamous cell carcinomatumor microenvironmentsingle cell RNA sequencingmalignant cellapoptosisCTSC |
| spellingShingle | Xin Sui Yongxu Jia Jing Li Jiayao Xu Wenjia Wang Yanru Qin Identification of CTSC-driven progression in ESCC by single-cell sequencing and experimental validation Frontiers in Immunology esophageal squamous cell carcinoma tumor microenvironment single cell RNA sequencing malignant cell apoptosis CTSC |
| title | Identification of CTSC-driven progression in ESCC by single-cell sequencing and experimental validation |
| title_full | Identification of CTSC-driven progression in ESCC by single-cell sequencing and experimental validation |
| title_fullStr | Identification of CTSC-driven progression in ESCC by single-cell sequencing and experimental validation |
| title_full_unstemmed | Identification of CTSC-driven progression in ESCC by single-cell sequencing and experimental validation |
| title_short | Identification of CTSC-driven progression in ESCC by single-cell sequencing and experimental validation |
| title_sort | identification of ctsc driven progression in escc by single cell sequencing and experimental validation |
| topic | esophageal squamous cell carcinoma tumor microenvironment single cell RNA sequencing malignant cell apoptosis CTSC |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2025.1585139/full |
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