Abstract: ObjectiveTo conduct a recombinant adeno-associated virus-2 carrying Slug-siRNA (rAAV2-Slug-siRNA),and to study the effect and mechanism of Slug-siRNA on AsPC-1 cells. MethodsThe targeting sequence of Slug gene which can be effectively silenced in RNA inference was confirmed in our previous study. The DNA containing both sense and antisense Oligo DNA fragments of the targeting sequence was designed, synthesized and cloned into the pDC316-EGFP vector. The insert was confirmed by PCR and sequencing. EGFP-Slug-siRNA sequence was amplificated and PCR product was obtained by EcoRⅠand SalⅠ digestion. pSNAV2.0-lacz-α plasmids was digested,and ligated with EGFP-Slug-siRNA. Transformation of DH5α with recombinant plasmids and identification of bacterial colonies containing recombinant plasmids by LB-agar plate were conducted，and pSNAV2.0-EGFP-Slug-siRNA were extracted purified and yerified. pSNAV2.0-EGFP-Slug-siRNA was transfected into BHK cells by means of lipofectamine. Using G418 Selection from mixed cells，BHK-Slug-siRNA was isolated，which was capable of Slug-siRNA expression,and was subsequenily infected with recombinant herpes simplex virusl(HSV1-rc/ΔUL2),which was able to pack the rAAV2-Slug-siRNA to form a functional and infectiou virus. After purifiation，the packaged rAAV2-Slug-siRNA was obtained.Western blot and RT-PCR methods was used to detect the expression of slug and PUMA after being infected AsPC-1 cells by Slug-siRNA for 48h,MTT and Annexin V/EGF methods was used to detect hibition rate and apoptosis of AsPC-1 cells. Results The recombinant adeno-assoeiated virus-2 vector carring Slug-siRNA was constructed successfully, using RT-PCR and restriction anzyme digestion，the destination gene was dectected. BHK-Slug-siRNA was was subsequenily infected with recombinant herpes simplex virusl(HSV1-rc/ΔUL2) able to pack the rAAV2-Slug-siRNA to form a functional and infectiou virus.The recombine viral titer was 9.23×1010(puf).Slug-siRNA inhibited the expression of Slug, at the same time, the proliferation of AsPC-1 cells was suppressed and the apoptosis was increased,significantly. Otherwise, with the decrease of Slug,the expression of PUMAwas increased. ConclusionThe constructed，high titer，recombinant adeno-associated virus-2 vector carring Slug-siRNA was constructed successfully.RNAi-mediated Slug gene with rAAV silencing could transfect to pancreatic cancer cells and silence Slug effectively and selectively, which resulted in the growth and proliferation inhibition in pancreatic cancer cells. The mechanism of Slug silencing in the growth and proliferation inhibition may be through upregulating expression of PUMA.