RNA interference suppresses expression of extraneous PC-1 gene in NIH3T3 cells

Objective: To study the correlation between PC-1 gene and prostate cancer by RNA interference technology, the most appropriate RNA interference target to PC-1 gene needs to be selected from candidates. Methods:Recombinant plasmids expressing short hairpin RNA targeting PC-1 mRNA were constructed using DNA recombinant technology and transfected into NIH3T3 cells with PC-1-EGFP plasmid respectively. The most effective RNA interferencing construct was selected from 5 candidates by observing the expression of EGFP using fluorescence microscope 48 hours after transfection. The effect of RNA interference was further confirmed by reverse transcriptase polymerase chain reaction and Western-blotting analysis. Results:PC-1 gene expression was surpressed by the most effective RNAi target. Conclusion:This strategy is adapted to select the most effective target from many candidates. The work provides basis for researhing the impact of suppression of the endogenous PC-1 gene expression on the biological behavior of prostate cancer cell.