短发夹RNA对胰腺癌细胞PANC-1突变型K-ras基因表达的影响

AIM: To study the specific inhibitory effect of short hairpin RNA (shRNA) on the mutant K-ras gene expression in PANC-1 cells. METHODS: Plasmid pGensil-1 recombinant plasmids targeting mutant K-ras gene was established. After transfection into PANC-1 cells, the expression of K-ras gene was detected using RT-PCR, Western blot techniques, and the effect of cell proliferation was determined by CCK-8 test. And a tumor model of nude mouse was constructed through rejecting the recombination plasmids to observe the volume of tumor. RESULTS: The recombinant plasmid encoding mutant-specific shRNA for K-ras inhibited the expression of mutant K-ras significantly (P=0.01). Rates of K-ras protein expression in non-treatment group, negative control group and treatment group were 100%, 99.0%±0.73%, 39.9%±2.1%, respectively. No significant difference in K-ras protein expression was observed between non-treatment group and negative control group. A 60.1% reduction in K-ras protein expression from the treatment group to non-treatment group indicated that cell growth was significantly inhibited (P=0.02); and after four-teen day's recombinant plasmid treatment, the volume of tumor grew slowly in nude mice and was significantly decreased compared with the control group (P=0.03). CONCLUSION: The specific shRNA inhibits the K-ras expression efficiently, and the growth of cells and nude mice is inhibited.