Silence Effect of Short Hairpin RNA on Expression of PIM-1 in Prostate Cancer Cell Lines / 天津医药

ABSTRACT

Objective: To investigate the interference effect of lipofectamine-mediated short hairpin RNA (shRNA) on the expression of PIM-1 gene in prostate cancer cell lines. Methods: Three kinds of recombinant plasmid expression vectors (pP[M]- hRNA-1,-2,-3) were constructed, which can produce shRNA targeting different sequence of PIM-1 mRNA. The recombinant plasmid vectors were transfected into PC-3 cell lines, respectively. RT-PCR and Western blot assay were used to detect the expression of PIM-1 mRNA and protein after transfection. Results: The expression of mRNA and protein of PIM-1 in PC-3 were inhibited significantly at 48h post-transfection. Furthermore, pPLM1- hRNA-2 and pPIM1- hRNA-3 resulted in a stronger silencing effect than that of pPIM- hRNA-1. Conclusion: The recombinant plasmid expression vector which carrying PIM-1 shRNA may remarkably inhibit the expression of PIM-1 mRNA and protein in PC-3 cell lines. The experiment provided the basis for further investigation of the role of PIM-1.