Inhibition Of CD147 Gene Expression Via RNA Interference Reduces Tumor Cell Invasion, Tumorigenicity And Increases Chemosensitivity To Gemcitabine In Pancreatic Cancer Cell Line Panc-1 Cells

Objective To investigate the effects of shRNA transfection on CD147 expression and cell proliferation and the potential of invasion and chemosensitivity to Gemcitabine in vitro and tumorigenicity in nude mice.Methods 1. The recombinant plasmid pSilencer-CD147shRNA was stably transfected into Panc-1 cells, and the mRNA and protein expression of CD147 was monitored by real-time PCR and Western blot.2. The change of proliferation level of the Panc-1 cells at 24, 48 and 72h after transfection and the chemosensitivity to Gemcitabine in vitro was detected by MTT.3. The effect of CD147shRNA on the level of MMP-2 and MMP-9 was detected by Gelatin zymography.4. The effect of CD147shRNA on invasion of the Panc-1 cells in vitro was detected by Transwell chamber.5. The effect of CD147shRNA on potential of tumorigenicity in nude mice was observed decreased.Results 1. The mRNA and protein expression of CD147 was significantly down-regulated after the Panc-1 cells was stably transfected by pSilencer-CD147shRNA, the interference of two shRNA target different sites of CD147 were 80.81% (P<0.05) and 34.58% (P<0.05) respectively, and the best inhibition clone was chosen to continue next study.2. The proliferation level of the Panc-1 cells at 24, 48 and 72h after transfection decresed to: 69.99% (P<0.05), 70.95% (P<0.05) and 70.61% (P<0.05) respectively and the chemosensitivity to Gemcitabine in vitro increased significantly (P<0.05).3. The level of MMP-2 and MMP-9 decresed significantly (P<0.05) after the Panc-1 cells were stably transfected by pSilencer-CD147shRNA.4. CD147shRNA transfection down-regulated the invasion in the Panc-1 cells in vitro significantly (P<0.05).5. CD147shRNA transfection decreased the size of subcutaneous tumor significantly (P<0.05).Conclusions 1. We successfully screened out pSilencer-CD147shRNA2 which has the best gene silence effect.2. We observed that that CD147 expression was significantly impeded at both mRNA and protein levels, which resulted in a decrease of the cell proliferation, MMP-2 and MMP-9 activities and invasive potential of Panc-1 cells as well as increased chemosensitivity to Gemcitabine in vitro and tumorigenicity in nude mice.