Objective：To evaluate the inhibitory effect of adenovirus-mediated HS-CRM8-survivin-EHV4 TK suicide gene on the xenograft tumor of hepatocellular carcinoma（HCC）in nude mice. Methods：A liver-specific adenovirus vector，Ad-CRM8-survivin-EHV4 TK（EHV4 TK），was constructed by molecular cloning and recombination. In nude mice bearing human hepatocellular carcinoma，the recombinant adenovirus was injected directly into the tumor followed by intraperitoneal injection of the prodrugs GCV and the inhibition effect on tumor xenografts was observed. The tumor section was stained with hematoxylin and eosin. Results： After adenovirus treatment, the volume of the tumors in EHV4 TK and GCV+EHV4 TK groups were（108.5 ± 18.3）and（37.6 ± 8.7）mm3. GCV+EHV4 TK represented much stronger anti-tumor effect on tumor growth than EHV4 TK only. GCV+EHV4 TK also showed much more tumor necrosis than EHV4 TK only. Conclusion：The adenovirus-mediated HS-CRM8-survivin-EHV4 TK suicide gene combined with GCV treatment may provide us a more effective，specific and convenient gene therapy method.