3D-bioprinted liver cancer models derived from primary hepatocytes for simulating cancer initiation and drug screening.

BACKGROUND: HCC remains one of the most lethal cancers globally, and accurately replicating the early events of tumor evolution remains a critical challenge. METHODS: In this study, we developed early-stage liver cancer cell lines by introducing distinct combinations of oncogenes into primary mouse hepatocytes. Using 3D bioprinting technology, combined with bioinks composed of gelatin and alginate, we constructed a more precise representation of liver cancer tissue to better simulate key tumor characteristics. RESULTS: Our·findings revealed that different oncogene combinations produced unique drug response profiles, with Ras-driven cells exhibiting heightened sensitivity to ferroptosis. Furthermore, 3D bioprinting tumor tissues derived from transformed hepatocytes effectively captured early HCC characteristics. These models preserved key features of early-stage liver cancer and provided a reliable platform for drug screening. Importantly, the 3D models demonstrated higher resistance to chemotherapy and targeted therapies compared with 2D cultures. CONCLUSIONS: In summary, we established both 2D and 3D models that replicate early HCC progression, offering valuable tools for drug screening and advancing our understanding of early carcinogenic mechanisms.