Objective: To investigate the effect of RNAi-mediated Survivin gene with conditionally replicating adenovirus silencing on Survivin gene in colon carcinoma cell lines HT-29 lastingly. Methods: We transfected Ad-delE1b55KD-shRNA / Survivin-EGFP to HT-29 (control was replication defective adenovirus and liposome vector which was contained the same shRNA as Ad-delE1b55KD-shRNA / Survivin-EGFP). The expressions of EGFP, Survivin mRNA and Survivin protein in HT- 29 were detected at the 1st, 7th, 14th and 28th days after transfection. Results: The expression of EGFP, the inhibition of Survivin mRNA and Survivin protein in HT-29 were high in each group at the 7th day after transfection, among the total, the effect of Ad-delE1b55KD-shRNA / Survivin-EGFP group was the highest; at the 14th day, the effects of replication defective adenovirus group and liposome vector group were decreased obviously, and it was still high in Ad-delE1b55KD-shRNA / Survivin-EGFP group; at the 28th day, the effects of control groups were disappeared, and it was still high in Ad-delE1b55KD- shRNA / Survivin-EGFP group like before (P < 0.05). Conclusion: RNAi-mediated Survivin gene with conditionally replicating adenovirus can silence Survivin gene in colon carcinoma cell lines HT-29 lastingly. Objective: To investigate the effect of RNAi-mediated Survivin gene with conditionally replicating adenovirus silencing on Survivin gene in colon carcinoma cell lines HT-29 lastingly. Methods: We transfected Ad-delE1b55KD-shRNA / Survivin-EGFP to HT-29 replication defective adenovirus and liposome vector which was contained in the same shRNA as Ad-delE1b55KD-shRNA / Survivin-EGFP). The expressions of EGFP, Survivin mRNA and Survivin protein in HT-29 were detected at the 1st, 7th, 14th and 28th days after transfection. Results: The expression of EGFP, the inhibition of Survivin mRNA and Survivin protein in HT-29 were high in each group at the 7th day after transfection, among the total, the effect of Ad-delE1b55KD-shRNA / Survivin-EGFP group was the highest; at the 14th day, the effects of replication defective defective adenovirus group and liposome vector group were significantly obviously, and it was still high in Ad-delE1b55KD-shRNA / Survivin-EGFP group; at the 28th day, the effects o f control groups were disappeared and it was still high in Ad-delE1b55KD-shRNA / Survivin-EGFP group like before (P <0.05). Conclusion: RNAi-mediated Survivin gene with conditionally replicating adenovirus vaccine silence survivin gene in colon carcinoma cell lines HT-29 lastingly.