To develop a ferroptosis-related prognostic model for esophageal squamous cell carcinoma (ESCC) using single-cell RNA sequencing and bulk transcriptomic data, addressing the need for improved prognostication in this aggressive malignancy.
Approach:
Data Integration: Integrated single-cell RNA sequencing and bulk transcriptomic data to identify ferroptosis-active cellular subpopulations, enhancing understanding of tumor microenvironment dynamics.
Prognostic Model Construction: Constructed a ferroptosis-related prognostic model using LASSO-Cox regression, validated across independent cohorts to ensure robustness.
In Vitro Experiments: Conducted functional experiments in vitro with ESCC cell lines to validate the prognostic core genes, providing insights into their biological roles.
Western Blot Analysis: Examined expression levels of ferroptosis-related proteins following CDCA3 knockdown to elucidate the molecular mechanisms involved.
Key Findings:
Identified four ferroptosis-related genes (CBS, CDCA3, GALNT14, IDO1) that stratified patients into high- and low-risk groups, with significant implications for survival outcomes.
CDCA3 knockdown inhibited proliferation and migration of ESCC cells and induced ferroptosis, highlighting its potential as a therapeutic target.
The identified genes were significantly upregulated in tumor cells at single-cell resolution and validated in clinical ESCC tissue samples, reinforcing their relevance in patient stratification.
Knockdown of CDCA3 led to downregulation of ferroptosis inhibitor-related genes and upregulation of ferroptosis-promoting genes, suggesting a shift in cellular response.
Interpretation:
The study presents a single-cell-resolved ferroptosis gene signature, contributing to the understanding of ESCC biology.
Limitations:
The study relies on data from specific cohorts and may not be generalizable to all ESCC populations.
Further clinical validation is necessary to confirm the utility of the prognostic model.
Conclusion:
The ferroptosis-related gene signature provides a foundation for developing targeted therapeutic strategies in ESCC.
