Background/Aims: The purpose of this research was to investigate the application of carbon dioxide (CO2) ablation and determine if carbon dioxide plays a role in treating hepatocellular carcinoma. Methodology: Twelve mice with subcutaneous xenografts of a hepatocellular cancer cell line were randomly separated into 2 groups. One group underwent CO2 ablation with their liver tumor in vivo while the other group in vitro, respectively. All mice were killed on day 28. Hematoxylin-eosin (HE) staining was performed on the mice to estimate the pathological change. Results: Just after 4 weeks, no significant difference in tumor size was detected between the two groups. The dimensions of the principal tumor varied from 2-3cm with an average size of 2.2cm in the greatest dimension. HE staining demonstrated an array of construct damage and necrosis in both groups. Conclusions:CO2 ablation could be an important treatment in the management of cancer.