Experimental study on lentiviral vector-mediated RNA interference inhibiting XIAP expression and influencing proliferation and apoptosis of human pancreatic cancer cells.

Objective:To investigate the possibility of XIAP inhibition by Lentiviral vector-mediated RNA interference and the influence on cell proliferation and apoptosis in pancreatic cancer cell line ,and to set up a pancreatic cancer cell line in which XIAP expression is stably suppressed. Methods: The Lentiviral vector of SiRNA targeted against XIAP (LV-XIAP-1, LV-XIAP-2,LV-XIAP-3) was constructed and transfected into the packagingcells 293T, and then collected supernatant with virus to transfect SW1990 cells. After seledtion by puromycin and culture expansion, the stable cell clones were attained; quantitative real-time fluorescent PCR and western-blot was used to detect the expression of XIAP. The effect of suppressing XIAP by RNAi on cell proliferation was quantified by methylthiazoletetrazolium (MTT) assay. Enzymatic activity of caspase-3/7 detecting and DAPI staining were employed to examine the apoptosis. Results: Three lentiviral vector-xiap-shRNA were constructed successfully, the xiap expression was decreased more than 70% in RNA and protein level when compared to contral. a pancreatic cancer cell line in which XIAP expression is stably suppressed was successfully set up. MTT showed that the cell number of XIAP expression suppression cell clones decreased significantly (P0.05), but the activity of caspase-3/7 and apoptosis index did not increase significantly. Conclusions: Lentiviral vector-mediad RNA interference targeting against XIAP can effectively inhibit XIAP expression and cell proliferation. The pancreatic cell line in which xiap gene was stablely suppressed was successfully established, which paves a way for continuous studying of XIAP function and gene therapy.