The Construction Of SMO Gene SiRNA Lentiviral Vector And Its Effect On SMO Gene Expression In Pancreatic Cancer Cells

Objective: To construct SMO targeting siRNA lentiviral expression vector, and confirmed inhibition to SMO gene expression of pancreatic cancer cells. Method:Designed and synthesized three SMO gene-targeted siRNA fragments, use recombinant DNA technology to build this three fragments carrying lentiviral expression vector and determination of the titer, after transfected the constructed lentiviral vector into human pancreatic cancer cell line SW1990, using RT-PCR method to detect the expression of SMO gene to filter out the best interference expression vectors, and detect the SMO protein by Western blot after transfected SW1990 cells. Result :Gene sequencing and restriction endonuclease digestion results suggest that successful synthesis of siRNA targeting SMO fragment and insert it into lentiviral vector correctly, non-base deletion staggered and mutation, determination of the virus titer is 5.31 × 108 TU / ML, 1.49 ×109TU / ML and 8.50 × 108 TU / ML, the transfected SW1990 cells, determination of the SMO gene expression inhibition rate is 86.00%, 74.85% and 19.22%, respectively, SMO protein inhibition rate> 80% after transfected the best Interference expression vector into SW1990 cells. Conclusion: Successfully constructed carry SMO gene-targeted siRNA lentiviral vector, and the vector can effectively inhibit the expression of SMO gene in pancreatic cancer cell,provides a good experimental tool for further research.