Objective: The investigate the inhibitory effect of the adenovirus-mediated CD/TK double suicide gene system driven by KDR promoter (Ad-KDRP-CD/TK) combined with survivin gene interference on the growth of hepatocellular carcinoma (HCC) cells in nude mice. Methods: Twenty nude mice were equally randomized into model group (subcutaneous implantation of HCC BEL-7402 cells to establish xenograft tumor without other additional treatment), double suicide gene transfection group (subcutaneous implantation of BEL-7402 cells transfected with Ad-KDRP-CD/TK, followed by intratumor injection with the prodrug gancilovir and 5-fluorocytosine after tumor formation), survivin siRNA transfection group (subcutaneous implantation of BEL-7402 cells, followed by intratumor injection with survivin siRNA/Lip-DMEM transfection complex after tumor formation), and combination transfection group (double suicide gene transfection plus survivin siRNA transfection). Two weeks after transfection treatment, mice in each group were sacrificed, tumor weight and tumor inhibition rate were measured, the microvessel density (MVD), and survivin mRNA and protein expressions in the tumor tissues were determined. Results: In each treatment group compared with model group, the tumor weight was significantly reduced, with the maximum tumor inhibition in combination transfection group (all P<0.05); the MVD, and the expression levels of survivin mRNA and protein were significantly decreased, with the maximum decreasing amplitude in combination transfection group (all P<0.05). Conclusion: Double suicide gene combined with survivin gene interference is an effective method to inhibit the growth of HCC cells in vivo.