TOP2A Drives Hepatocellular Carcinoma Progression: Integrative Multi-Omics Analysis Reveals Prognostic Power and Actionable Mechanisms

Objective To investigate the biological functions of TOP2A in hepatocellular carcinoma (HCC) using bioinformatics and single-cell RNA sequencing (scRNA-seq) methods, and to explore the epigenetic regulatory mechanisms of TOP2A in HepG2 and Huh-7 cells with TOP2A knockdown. This study aims to provide new theoretical insights and potential therapeutic targets for the early diagnosis and targeted treatment of HCC. Methods The sample information of liver cancer was sourced from TCGA and GEO databases, while the epigenetic gene data were from EpiFactors database. Single-factor Cox regression was employed to screen the prognostic genes from TCGA and GEO. By merging with the EpiFactors database, epigenetic-related genes ERGs that affected prognosis were obtained and enrichment analysis was conducted on them. Subsequently, a multivariate Cox regression was used to construct a prognostic model, nomogram was drawn and validation was performed. Through the analysis of single-cell RNA sequencing data, the immunological significance of TOP2A and its relationship with the diagnosis and prognosis of liver cancer were studied.The expression levels of TOP2A in HCC and normal liver epithelial cells were validated using quantitative PCR (QPCR). siRNA technology was employed to interfere with TOP2A expression in HepG2 and Huh7 cells, and a TOP2A knockdown cell line was established. The transfection efficiency of siRNA was measured using QPCR. The impact of TOP2A knockdown on the proliferation of HCC cells was assessed using the CCK8 assay. The effect of TOP2A on the migration ability of HCC cells was observed through a wound-healing assay. The influence of TOP2A knockdown on the invasive ability of HCC cells was examined using the Transwell assay. The effect of TOP2A knockdown on the apoptosis of HCC cells was evaluated using flow cytometry. Results We constructed a prognostic model based on three ERGs. The Kaplan-Meier curve demonstrated that the prognosis of the high-risk group was worse than that of the low-risk group.