Construction of Eukaryotic Expression Vector for Survivin-siRNA and Its Effect on Proliferation and Apoptosis of Gastric Cancer SGC-7901 Cells

Objective To construct a eukaryotic expression vector for Survivin-siRNA and investigate its effect on proliferation and apoptosis of gastric cancer SGC-7901 cells.Methods Four single-stranded template DNAs encoding siRNA against Survivin,each consisting of 75 bp,were synthesized chemically,based on which two double-stranded DNAs were formed by annealing,then identified by restriction analysis and inserted into vector pSUPER.basic.SGC-7901 cells were transfected with positive recombinants,then counted and determined for proliferation activity by MTT method,and for cell cycle by flow cytometry.The transcription level of Survivin mRNA was determined by semi-quantitative RT-PCR.Results Both PCR and restriction analysis proved that the eukaryotic expression vector for Survivin-siRNA was constructed correctly.The constructed expression vector down-regulated the transcription level of Survivin mRNA,inhibited the growth and proliferation and promoted the apoptosis of SGC-7901 cells.The number of transfected SGC-7901 cells at G0 /G1 and sub-G1 stages increased,while that at S stage decreased.Conclusion The eukaryotic ex-pression vector for Survivin-siRNA was successfully constructed,which down-regulated the transcription of Survivin mRNA in SGC-7901 cells,and inhibited the proliferation and promoted the apoptosis of the cells.It provided a certain experimental basis for gene therapy of gastric cancer by RNAi technique.