Supplementary Material for: EMT-Related Gene Signature: A New Method for Personalized Risk Assessment in Patients with Hepatocellular Carcinoma

Background: Hepatocellular carcinoma (HCC) is the fourth-leading cause of cancer-related death worldwide, accounting for more than 700,000 deaths annually. Epithelial–mesenchymal transition (EMT) is posited to contribute to HCC progression. We therefore aimed to elucidate the relationship between EMT-related gene (ERG) expression and prognosis in patients with HCC. Methods: Univariate and multivariate Cox regression analyses were performed to identify prognostic-related differentially expressed EMT-related genes (DE-ERGs). Time‑dependent receiver operating characteristic (ROC) and Kaplan–Meier curves were used to assess the prognostic performance of the EMT-related signature. pRRophetic R package was used to evaluate sorafenib sensitivity in the GSE14520 cohort. Gene expression in Huh7 and L02 cell lines was detected by real-time PCR and western blotting. Differential expression of the three genes between HCC tissues and normal tissues was validated using immunohistochemical analyses. Results: Of the 76 identified DE-ERGs, 29 were associated with overall survival. Three prognosis-related ERGs (GOLM1, SOX4 and CD14) were screened out by multivariate Cox regression. A gene signature was identified based on the three prognostic related ERGs. HCC patients with a low-risk score had a better prognosis and were more sensitive to sorafenib compared to those with a high-risk score. Moreover, we further confirmed increased expression of GOLM1 and SOX4, and decreased expression of CD14, in liver cancer cell line and HCC tissue. Conclusions: The results of the present study demonstrate the utility of an EMT-related gene signature as a potential biomarker informing prognosis in patients with HCC, which may contribute to the implementation of personalized therapies.