Unveiling the Therapeutic Potential of Targeting RRM2 in Hepatocellular Carcinoma: An Integrated In Silico and In Vitro Study.

Hepatocellular carcinoma (HCC) has attracted global and local interest, with alarming current and forecasted statistics. This is mainly due to its late-stage diagnosis, high recurrence rate and potential resistance to conventional therapies. Thus, there is an emerging need to explore new molecular targets and novel treatment strategies to effectively combat HCC. In this study, we analyzed four GEO datasets (GSE112790, GSE62232, GSE60502 and GSE84402) to identify upregulated differentially expressed genes (DEGs) associated with HCC. Enrichment analysis and protein‒protein interaction (PPI) network construction were performed on the overlapping upregulated DEGs to predict and prioritize potential therapeutic targets. Six hub genes (RRM2, TOP2A, CCNB1, CDK1, BIRC5 and PBK) were identified, with RRM2 emerging as the top candidate. The role of RRM2 in HCC was then validated through a literature review, in silico analysis and CRISPR/Cas9-mediated gene disruption, followed by in vitro functional assays in HepG2 cells. RRM2 knockdown resulted in significant reductions in cell viability, proliferation inhibition, migration impairment, G2/M phase cell cycle arrest and apoptosis. Our findings emphasize RRM2 as a critical player in HCC progression. Targeting RRM2 with CRISPR/Cas9 effectively reversed several hallmarks of HCC, offering new opportunities for the development of precise HCC therapeutic options. GRAPHICAL ABSTRACT: [Image: see text] SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10142-025-01630-0.