The effect of antisense oligonucleotide (ASODN) targeting survivin on renal cell carcinoma (RCC) apoptosis, proliferation and sensitivity to chemotherapeutic drugs was investigated. Antisense oligonucleotide targeting survivin was designed and composed. The cDNA of survivin was transfected into RCC cell lines ACHN and 769-P, respectively. They were both cultured in normal condition. After transfection, ACHN and 769-P cell lines were cultured in a 6-well culture plate. The cultured cells were divided into 6 groups. Twenty-four h after transfection, survivin protein expression was detected by Western blot. Apoptotic index (AI) and proliferative index (PI) were examined by flow cytometry. Rate of inhibition (IR) by chemotherapeutic drugs was determined by the colorimetric MTT cell viability/proliferation assay. The results showed AI and IR of chemotherapeutic drugs in ASODN groups were significantly higher than those in the groups without ASODN. There were statistical differences between ASODN groups and control groups (P< 0.05), but there was no difference among control groups (P>0.05). The PI in the ASODN groups was significantly lower than that in the control groups with the difference being statistically significant (P<0.05). In addition, there were no differences among control groups. It was concluded that the expression of survivin could be down-regulated in 2 cell lines after ASODN transfection. Antisense oligonucleotide targeting survivin could induce RCC apoptosis, inhibit cell proliferation and sensitize RCC to chemotherapy.