Objective：To construct survivin shRNA expression vector and investingate the effect of survivin shRNA on chemotherapy resistance of GBC-SD cells.
Methods：The siRNA sequence targeting survivin mRNA was synthesized and cloned into pEGFP-H1. The constructed plasmid and pEGFP-H1 were transfected into GBC-SD cells respectively via liposome. Then the transfected cells were selected with G418. GBC-SD cells were divided into three groups: GBC-SD, GBC-SD/EGFP and GBC-SD/survivin groups. Survivin mRNA was tested by RT-PCR. Then cells of the 3 groups were treated with adequate concentration of DDP（3.0 μg/mL） for similar periods of time, cell survival rate was detected with MTT and apoptosis was observed by TUNEL.
Results：The recombinant plasmid, pEGFP-survivin, was successfully constructed.  Compared to the other 2 groups, the level of  survivin expression in GBC-SD/survivin group was obviously decreased (74.7%, 71.5%). After DDP treatment, cell survival rate was obviously decreased in GBC-SD/survivin group compared with other 2 groups. There were brownly apopotosis nucleuses in the three groups.
Conclusions：Survivin shRNA expression vector has been constructed successfully and GBC-SD cells with stable expression shRNA has been obtained. The survivin shRNA could significantly down-regulate the expression of survivin in GBC-SD cells and improve the sensibility to chemotherapy.