Identification of CTHRC1 as a Potential Prognostic Biomarker in Esophageal Squamous Cell Carcinoma by Bioinformatics Analysis

Background Esophageal squamous cell carcinoma (ESCC) carries one of the highest case-fatality rates among gastrointestinal malignancies, owing in part to the scarcity of molecularly validated prognostic indicators. Collagen triple helix repeat containing 1 (CTHRC1), a secreted glycoprotein implicated in matrix remodeling across multiple tumor types, has yet to be systematically interrogated in ESCC. The present investigation aims to define the prognostic relevance of CTHRC1 in ESCC through a multi-layered bioinformatics framework. Methods Transcriptomic profiles from the TCGA-ESCA project and two independent GEO cohorts (GSE53624, GSE23400) were leveraged to quantify CTHRC1 abundance relative to normal esophageal mucosa. Weighted gene co-expression network analysis (WGCNA) and protein–protein interaction (PPI) mapping were applied to delineate CTHRC1-centric functional modules and molecular interaction hubs. Single-cell RNA sequencing (scRNA-seq) data from GSE160269 were interrogated to resolve cell-type-specific CTHRC1 distribution within the tumor ecosystem. Prognostic relevance was appraised via Kaplan–Meier survival curves, Cox regression, and time-dependent ROC analysis. Experimental corroboration was obtained by qRT-PCR and immunofluorescence (IF) in a panel of ESCC lines (EC109, KYSE-150, KYSE-30, TE-1) and primary cancer-associated fibroblasts (CAFs), supplemented by CTHRC1 loss- and gain-of-function manipulation in KYSE-30. Results Across all three genomic cohorts, CTHRC1 transcripts were markedly elevated in tumor versus matched normal tissue (p < 0.001). WGCNA assigned CTHRC1 to a gene module enriched for extracellular matrix (ECM) organization, epithelial–mesenchymal transition (EMT), and invasive signaling, while PPI mapping disclosed functional links to collagen family members and matrix metalloproteinases. Single-cell profiling localized peak CTHRC1 activity to cancer-associated fibroblasts and a defined subset of malignant epithelial cells. Clinically, elevated CTHRC1 tracked with higher T and N classifications and advanced pathological stage; high expressors faced substantially inferior overall survival (multivariate-adjusted hazard ratio = 1.78, 95% CI: 1.23–2.57, p = 0.002), with multivariate Cox analysis confirming its independence from conventional staging parameters. In vitro, qRT-PCR revealed 4.8–8.5-fold CTHRC1 mRNA enrichment across all four ESCC lines relative to HEEC (p < 0.001). IF experiments demonstrated cancer-restricted CTHRC1–Pan-CK co-localization, CTHRC1-dependent modulation of Vimentin and MMP9 abundance, and selective CTHRC1–α-SMA co-expression in myofibroblastic CAFs. Conclusions Integrative multi-omics interrogation nominates CTHRC1 as a reproducibly overexpressed, clinically actionable prognostic molecule in ESCC. Its tight coupling to ECM remodeling pathways, tumor-stromal crosstalk, and unfavorable survival outcomes positions CTHRC1 as a candidate for risk-stratification algorithms and targeted therapeutic development. Prospective multi-center validation and mechanistic dissection in vivo remain essential next steps toward clinical translation.